Review Article Recent Advances in Diagnosis, Prevention

Hindawi Publishing CorporationAdvances in VirologyVolume 2013 Article ID 595768 26 pageshttpdxdoiorg1011552013595768

Review ArticleRecent Advances in Diagnosis Prevention and Treatment ofHuman Respiratory Syncytial Virus

Swapnil Subhash Bawage Pooja Munnilal Tiwari Shreekumar PillaiVida Dennis and Shree Ram Singh

Center for NanoBiotechnology Research Alabama State University Montgomery AL 36104 USA

Correspondence should be addressed to Shree Ram Singh ssinghalasuedu

Received 31 July 2013 Accepted 30 September 2013

Academic Editor Subhash Verma

Copyright copy 2013 Swapnil Subhash Bawage et alThis is an open access article distributed under theCreativeCommonsAttributionLicense which permits unrestricted use distribution and reproduction in anymedium provided the originalwork is properly cited

Human respiratory syncytial virus (RSV) is a common cause of respiratory infection in infants and the elderly leading tosignificant morbidity and mortality The interdisciplinary fields especially biotechnology and nanotechnology have facilitated thedevelopment of modern detection systems for RSV Many anti-RSV compounds like fusion inhibitors and RNAi molecules havebeen successful in laboratory and clinical trials But currently there are no effective drugs for RSV infection even after decadesof research Effective diagnosis can result in effective treatment but the progress in both of these facets must be concurrent Thedevelopment in prevention and treatment measures for RSV is at appreciable pace but the implementation into clinical practicestill seems a challenge This review attempts to present the promising diverse research approaches and advancements in the area ofdiagnosis prevention and treatment that contribute to RSV management

1 Introduction

Worldwide there are reportedly about 12 million severeand 3 million very severe cases of lower respiratory tractinfection (LRTI) in children [1] Respiratory syncytial virus(RSV) is a common contributor of respiratory infectionscausing bronchiolitis pneumonia and chronic obstructivepulmonary infections in people of all ages but affects mainlychildren and elderly along with other viral infections leadingto highmortality andmorbidity [2ndash4] A recent global surveysuggests that RSV is not prevalent throughout the year inthe tropical regions of the globe but the incidence peaks inwinter with a wide ranging persistence depending on the geo-graphical topology [5] RSV has been reported to be a preva-lent lower respiratory tract pathogen distributed worldwideincluding countries from both the developed and developingworld The major countries with RSV seasonal outbreaksinclude USA Canada Cambodia Mexico Uruguay BrazilPeru France Finland Norway Sweden Latvia DenmarkGermany Netherlands Ireland Italy Turkey Iran SaudiArabia Australia New Zealand China Korea Hong KongJapan India Pakistan Bangladesh Nepal Taiwan Vietnam

Myanmar Thailand Madagascar Kenya Zambia Nigeriaand Columbia The data about human RSV described inliterature over the years seem to have been unchangedsignificantly indicating the severity of RSV and the urgentconcern to address this issue An estimate of more than 24billion US dollars per year is the economic cost of viral lowerrespiratory tract infection in children [6]

RSV is a Paramyxovirus belonging to the genus Pneu-movirus RSV is an enveloped nonsegmented negativesingle stranded linear RNA genome virus (Figure 1) RSVgenome (sim15 kb) has 10 genes encoding 11 proteins withtwo open reading frames of gene M2 [7 8] Other genesinclude nonstructural proteins NS1 and NS2 (type I inter-feron inhibitors) L (RNA polymerase) N (nucleoprotein) P(Phosphoprotein cofactor for L) M (Matrix protein) M21andM22 (required for transcription) SH (small hydrophobicprotein) G (glycoprotein) and F (fusion protein) Being anegative strand RNA genome virus RSV packages its ownpolymerase into the nucleocapsid Of these proteins fusionprotein (F) is indispensable for viral attachment to the hostand entry into the host cell Although the G protein isresponsible for the preliminary attachment the F protein is

2 Advances in Virology

G protein

N protein

F protein

(a) (b)

P protein L protein

SH protein

Single stranded RNA

Matrix protein

NS1

NS2

N

P

M

SH

G

F

M2

L

Figure 1 Structure and genome organization of respiratory syncytial virus (a) Approximately 200 nm RSV virion particle and (b) singlestranded negative RNA genome consisting of 10 genes

necessary for the fusion budding and spread of the virus[9 10] After attachment to the host cell RSV fuses withthe host cell membrane using the F protein through the6 helix coiled-coil bundle of the F protein a mechanismcharacteristically found in paramyxoviridae members [11]Although the detailed mechanism of RSV infection is notfully understood the most accepted mechanism is the entryof the nucleocapsid into the host cell mediated by the Fprotein through clathrin mediated endocytosis [12] TheRNA is first converted into a plus strand which serves asthe template for replication whereas for transcription theRNA genome itself transcribes mRNA for protein synthesiswithout any intermediate

Almost all children of 2 years of age will have had an RSVinfection and leading to 160000ndash600000 deaths per year [4]Approximately 25 to 40 of infants and children at thefirst exposure to RSV have signs or symptoms of bronchiolitisor pneumonia These symptoms include rhinorrhea low-grade fever cough and wheezing The symptoms in adultsmay include common cold with rhinorrhea sore throatcough malaise headache and fever It can also lead toexacerbated symptoms such as severe pneumonia in theelderly especially residing in nursing homes [13] Usuallychildren show symptoms within 4 to 6 days of infectionand most of them recover in 1 to 2 weeks while serving

as carriers of the virus for 1 to 3 weeks RSV infectionin children of nosocomial origin is associated with highermortality than community-acquired illness because of thepre-existingmorbidity [14 15] SevereRSVdisease risk hoversfor the elderly and adults with chronic heart or lung diseaseor with weakened immune system [16] RSV infection doesnot provoke lasting immunity [17] therefore reinfection isvery common [18] Recently RSV infection was reported toaccount for hospitalizations and mortality in elderly people[19] RSV accounted for severe lower respiratory tract infec-tions including chronic lung disease systemic comorbiditiesand even death At present there is no specific treatmentfor RSV infection ever since its first discovery in 1956 [20]Currently Food and Drug Administration (FDA) approvedprophylactic drug for RSV that includes palivizumab andribavirin administered along with symptomatic treatmentdrugs and supportive care Currently techniques used fordiagnosis of RSV include ELISA direct immunofluorescencewestern blot PCR and real-time PCR The diagnosis andtreatment scenario has significantly changed with the adventof advanced techniques and in-depth understanding of RSVbiology but the execution of these clinical developmentsin practice requires extensive study and time This reviewpresents the recent advances in the diagnosis prevention andtreatment of RSV (Figure 2)

Advances in Virology 3

Nanotechnology based assays

Quantum dots

Immuno-PCR

Microarray

Live cells RNA imaging

SERS

LFIA

ELISA

Immunoassays

OIA

DFA

LAMP

Real-time PCR

DNA based assays

PCR-ESI-MS

Microarray

DNA vaccines

Subunit vaccines

Prevention

AntibodiesNutrition

Nanovaccines

Fusion inhibitorsAnti-viral drugs

Treatment

Antisense RNA

EthnobotanicalsNanoparticles

Diagnosis

Figure 2 A schematic representation of RSV management through coordinated diagnosis prevention and treatment

2 Diagnosis

There are ten known genotypes of RSV based on the sequenceanalysis of the RSVgenomewhich suggests that the pathogenchanges with time and the resulting genotypes pose a threatto the public health Information from comparative genomeanalysis has been utilized for evolutionary studies and mostimportantly for the development of detection and treatmentstudies [21 22] Diagnostic assays for detection of pathogenshave a pivotal role in public-healthmonitoring [23]Howeverdiagnosis becomes difficult when the causative agent exhibitsoverlapping symptoms with other disease(s) or remainsnonsymptomatic Hence the correct diagnosis has to beempirically derived for the treatment Some of the promisingmolecular and biophysical techniques for RSV diagnosis arediscussed below (Table 1)

21 Immunoassays211 Enzyme-Linked Immunosorbent Assay Enzyme-linkedimmunosorbent assay (ELISA) is the enzyme facilitated col-orimetric detection of specific protein-antibody complexesELISA is extensively used for detection of various proteins atvery low concentration in different sample types thusmakingit clinically significant in routine diagnosis of pathogensELISA for RSV detection is mainly based on targeting RSVF protein (antigen) Recently several modifications of theclassical ELISA technique have been efficiently developedand employed for the detection of RSV Sensitivity of ELISAwas increased by using the high affinity anti-RSV F antibodypeptides derived from the motavizumab [85] Motavizumabis a high affinity antibody based therapeutic against RSVwhich binds to RSV F protein however it was disapproved bythe FDA due to higher hypersensitivity in patients receivingmotavizumab as compared to palivizumab Motavizumab

also caused urticaria [34] It is presumed to be a betterbinding target than the conventional F protein ELISA Thismethod could prove more effective than the F proteinELISA due to its higher sensitivity to the degradation ofmotavizumab In a simple thin layer amperometric enzymeimmunoassay RSV was detected as early as 25 minutes atlow cost with comparative sensitivity to that of real-timePCR and immunofluorescence assays [35]The assay is basedon the development of a double layer sandwich methodsimilar to ELISA It involves a polystyrene microarray slidecoated with monoclonal antibody which captures the antigen(RSV) The antigen-antibody complex is detected by horseradish peroxidase conjugated secondary antibody on a screenprinted electrochemical cell coated with the substrate

212 Immunofluorescence Assay Presently immunofluores-cence is one of the most common and rapid RSV detectiontechniques used where the antigen is detected by a fluo-rescently tagged antibody Direct fluorescent antibody assay(DFA) is a standard detection technique used for decadesand other RSV detection techniques are often comparedto DFA for evaluating their efficiency [24] An indirectassay which uses secondary fluorescently tagged antibody isanother option to DFA Currently there are many moleculartechniques that are more sensitive and reliable than DFAbut DFA is widely used for RSV detection in clinical samplesdue to the ease and rapidity The sensitivity and specificity ofDFA are a subject of variance as the success of the techniqueis dependent on numerous factors mainly the skills of thetechnician and nature of the sample A study showed thatDFA could detect RSV with a sensitivity and specificity of778 and 996 respectively (positive predictive value was986 and the negative predictive value was 94) DFA isa reliable technique for RSV detection for patients tested

4 Advances in VirologyTa

ble1Com

paris

onof

RSVdetectiontechniqu

es

Techniqu

eRe

ference

Principle

Advantages

Drawbacks

Currentu

sage

status

(A)F

luorescenceb

ased

metho

ds

(1)D

FA[2425]

Microscop

icdetectionof

RSVwith

specifica

ntibod

yconjugated

with

fluorop

hore

Easy

procedure

Hum

anerrorfading

ofdyes

Research

intentH

ospitalbased

procedurecommercial

diagno

sticassays

(2)Q

Ds

[26ndash

31]

Detectio

nof

signalsfro

mflu

orescent

nano

particles

upon

encoun

terw

ithRS

Veither

throug

hmicroscop

yor

flow

cytometry

Photostableinorganicinnatureresistant

tometabolicdegradation

Toxicityinsolub

ility

Research

intent

(3)M

olecular

beacon

based

imaging

[3233]

Hairpin

DNAfunctio

nalized

gold

nano

particlewith

fluorop

hore

hybridizationwith

targetmRN

ALive

cellim

agingwith

real-timed

etectio

nProb

ablegene

silencing

metabolicdegradation

Research

intent

(B)Immun

oassays

(1)E

LISA

[3435]

Specificb

inding

andcolorim

etric

detectionof

antig

en-antibod

ycomplex

Easy

protocolhighspecificityand

sensitivity

Cumbersom

epron

etohu

man

errors

Hospitalbased

procedure

commercialdiagno

sticassays

(2)O

IA[36ndash

38]

Presence

ofspecifica

ntigen-antibod

ycomplex

form

edaltersther

eflectiv

esurfa

cesp

ropertiesw

hich

isvisually

detected

Easyrapidspecificitycosteffective

Needs

confi

rmationby

other

tests

forn

egatives

amples

Research

intentnot

commercialized

(3)L

FIA

[39ndash

42]

Immun

o-complexes

detected

chromatograph

ically

EasyrapidhandycosteffectiveFD

Aapproved

Non

quantitativelim

itof

sample

volumelim

itsdetection

Hospitalbased

procedure

commercialdiagno

sticassays

(C)M

olecular

metho

ds

(1)L

AMP

[4143ndash4

5]Colorim

etric

turbidim

etric

detectionof

isothermalam

plificatio

nof

DNAusing

specificp

rimer

Sensitivityandspecificity

Semiquantitativ

edesig

ning

compatib

leprim

erset

Research

intentnot

commercialized

(2)P

CR[4647]

Amplificatio

nof

viralcDNAand

visualizationof

PCRprod

uct

Rapidandsensitive

than

conventio

nal

cultu

remetho

dsHighlim

itsof

detection

Research

intenthospitalbased

procedure

(3)R

eal-T

ime

PCR

[48ndash53]

Real-timea

mplificatio

nof

targetDNAor

cDNA

Rapid(3ndash5

hours)highlysensitive

and

very

lowlim

itsof

detection

Expensive

Research

intenthospitalbased

procedurecommercialassay

(4)M

ultip

lex

PCR

[54ndash

57]

Use

ofmultip

leprim

erandor

prob

esets

Simultaneou

sdetectio

nof

multip

lepathogenicspecieso

rstrains

Lesssensitive

Research

intenthospitalbased

procedure

(5)Immun

o-PC

R[5859]

Acombinatio

nof

immun

oassay

and

real-timeP

CR

Very

lowlim

itsof

detection

improved

limits

ofdetectionover

individu

alEL

ISA

andPC

R(400

0and4foldrespectively

)Com

plex

experim

entald

esign

Research

intentnot

commercialized

(6)M

icroarray

[60ndash

74]

Hybrid

izationof

sampleb

iomolecules

toim

mob

ilizedtargetDNAor

proteinon

achip

Highlysensitivelarge

scaleidentificatio

nof

multip

lepathogensproteinand

nucle

icacid

targets

Cost-ineffective

Research

intenthospitalbased


(D)B

ioph

ysicalmetho

d

(1)P

CR-ESI-M

S[7576]

Massspectroscop

yof

PCR-am

plicon

sthroug

hele

ctronspraydispersio

n

Highlysensitive

andspecifice

venat

strain

leveland

efficientm

ultip

lepathogensd

etectio

nEx

pensive

Research

intentnot

commercialized

(2)S

ERS

[77ndash84]

Inelastic

scatterin

gof

mon

ochrom

atic

radiationup

oninteractionwith

ananalytew

ithlow-fr

equencyvibrational

andor

rotatio

nalenergy

Rapidandno

ndestructiv

edetectio

nof

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with

high

sensitivity

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reparatio

nRe

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within first 3 days after onset of symptoms but the sensitivitydecreases if tested 4ndash7 days after the onset of the symptoms[24] DFA can detect RSV alone or as a multivalent testfor other respiratory viruses (influenza A and B virusesparainfluenza virus types 1 to 3 and adenovirus) usingSimulFluor Respiratory Screen assay [25]

213 Optical Immunoassay The direct visualization of anantigen-antibody complex often referred to as optical immu-noassay (OIA) is used for qualitative detection of Streptococ-cus and influenza virus Visualization of immunocomplexescan be enhanced by immobilizing themon a special reflectingsurface The antigen-antibody complex forms a thin layerwhich changes the reflective properties of the surface [36]The technique is simple rapid and sensitive and as low as1 ng of antibody per mL can also be detected [37] OIA isnow used for detection of RSV with a sensitivity specificitypositive and negative predictive values of 879 996989 and 945 respectively Though OIA offers rapid andcost-effective RSV detection it is recommended that negativeresults of OIA should be confirmed by other tests [38]

214 Lateral Flow Immunoassay Lateral flow immunoassay(LFIA) is an immunochromatographic technique known forthe rapid detection of RSV from nasal washes or nasalaspirates The lateral flow of antigen-antibody complex onthe substrate matrix reaches the reaction area and results information of colored band indicating the presence of antigenin the specimen There are many LFIA kits like Remel XpectBinax Now RSV BDDirectigen EZ RSV QuickLab RSV Testand RSV Respi-Strip [39ndash42] The sensitivity and specificityare normally above 90 and 95 respectively but vary asper the manufacturer Modifications of LFIA may includecolloidal gold conjugated with antibody specific to RSV inthe matrix for trapping the antigen and assisting the goldmediated reaction for color band development

22 Loop-Mediated Isothermal Amplification Loop-mediat-ed isothermal amplification (LAMP) is a nucleic acid baseddetection method used for bacterial and viral pathogens Itcan be used for RNA viruses where the additional step ofcDNA synthesis is required and commonly designated asreverse transcription LAMP (RT-LAMP) LAMP consists ofpairs of primers specific to cDNA which is then amplified byautocycling strand displacement activity of DNA polymerasegenerally at 60∘C The reaction time of 1 to 15 hours issufficient and can be monitored by real-time turbidimeterand the resulting product can also be viewed by agarosegel electrophoresis This method can distinguish RSV strainA and B upon restriction digestion of the product [41]Alternatively RSV strain A and B can be detected designingspecific primer sets for them The efficiency of RT-LAMPwas tested for RSV detection from nasopharyngeal aspiratesand compared with viral isolation enzyme immunoassayimmuno-chromatographic assay and real-time PCR RT-LAMP was the most sensitive among all the methods testedwith the exception of real-time PCR Also the RT-LAMPwasspecific for RSV and did not react with any other respiratory

pathogens like RNA viruses DNA virus or bacteria [43]Although this method is rapid the requirement of theturbidimeter makes it slightly inconvenient as a cost-effectivedetection technique Development in the chemistry of theLAMP has made it possible to get rid of the turbidimeter asit is now possible to visually detect the presence of pathogenin the test sample LAMP is now referred to as colorimetricdetection of loop-mediated isothermal amplification reactiondue to incorporation of the dye in the chemistry to aid visualdetection Wang et al (2012) have developed such an RT-LAMP assay for the detection of human metapneumoviruses(which include rhinovirus RSV influenza virus APR834(H1N1)) which can detect as low as ten viral RNA copieswith better efficiency than RT-PCR [44] It is now possibleto detect RSV A and B strains in nasopharyngeal specimenusing multiplex LAMP (M-LAMP) in just 30 minutes M-LAMP had a sensitivity and specificity of 100 when com-pared with PCR [45]

23 Polymerase Chain Reaction (PCR) Based Detection

231 Conventional PCR The applicability of PCR is ubiqui-tous and has made the diagnosis of pathogens tremendouslyrapid and sensitive [46] RSV is usually challenging to detectdue to poor viral titer and sensitivity to antigen based detec-tion methodsThus a PCR based method was developed andcompared to serological and culture based detectionmethodsin adults having respiratory infections [47]The PCRmethodwas based on the reverse transcription-nested PCR techniqueinvolving the outer and inner primers designed from theF gene of RSV strain A over a two day procedure with asensitivity of 73 This method is faster as compared to theconventional culture method which usually takes 3ndash5 daysresulting in faster treatments

232 Real-Time PCR Although conventional reverse tran-scriptase-PCR (RT-PCR) is sensitive as compared to theculture methods it suffers from the lower sensitivity Thisproblem has been obviated by the development of real-timePCR based methods Real-time PCR is by far the mostsensitive method for the detection and diagnosis of a widearray of pathogens including RSV Several studies have beenconducted for the development of real-time PCR assays forRSV detection especially the RT-PCR In one such studya rapid sensitive and specific method was devised basedon TaqMan real-time PCR for the detection of both RSV Aand RSV B strain [48] The primer and the probe sets weredesigned from the nucleocapsid gene (N) The sensitivity ofthismethodwas found to be 0023 PFU (plaque forming unit)or two copies of mRNA for RSV A whereas the sensitivityfor RSV B was 0018 PFU or nine mRNA copiesThis methodis fast and efficient as the diagnosis was performed within 6hours of the procurement of samples

A similar assay based on real-time PCR was developedand employed in the detection of RSV in the bronchoalveolarlavage (BAL) of lung transplant patients or patients withrespiratory infections [49] The assay was designed based onthe RSVN gene and first involved a screening step of the RSV


positive samples using a SYBR green based assay followedby quantitative real-time PCR using TaqMan based assay tobe more cost effective Also the assay was developed for thedetection of both RSV A and B subgroups separately Theassay was found positive in 16 of transplant patients thusindicating some possible association between RSV infectionand lung transplant tolerance

Real-time PCR based on RSV N gene has also been usedto quantify RSV from the nasal aspirates of children [50]Themethodwas one logmore sensitive than the conventionalculture method The real-time PCR assay resulted in 56 ofpositives whereas the immunofluorescence assay had 413and culture method had 453 of positives Surprisingly theRSV to GAPDH ratio did not differ in children with severe ornonsevere infection raising confusion about the correlationof viral load to the severity of infection due to the intensityof viral replication genetic susceptibility of the host andimmune responses thus making it necessary to considerother factors besides the viral load

A similar assay was developed for the detection of RSVusing primer-probe sets from the RSV F gene [51] A com-parative analysis wasmade between the nested RT-PCR real-time PCR and ELISA wherein real-time PCR was 25moresensitive than the conventional nested PCR thus making itmore applicable for clinical samples Additionally the real-time PCRwas performed in two steps to increase the sensitiv-ity of the samples This method was more rapid as the resultswere obtained in 35 to 4 hours upon receiving the samplesSeveral other reports also emphasize the applicability of real-time PCR in detection of RSV in immunocompromisedpatients [52] Besides the RSV F and N gene the RSV matrixgene and polymerase gene have also been used to detect RSVin children [53]The latter method had the ability to quantifyand classify RSV with high efficiency and rapidity Howeverthe sensitivity of the technique relies on the age of thepatients

233 Multiplex PCR Themultiplex PCR approach has beenused for the detection and subtyping of RSV and humaninfluenza virus simultaneously [54] and as many as 18respiratory viruses could be detected [55] The method wasbased on the primer sets of hemagglutinin and nucleoproteingene of influenza virus and nucleocapsid protein gene ofRSV and found to be rapid specific and sensitive Howeverthe method suffered from the drawback that it could notdistinguish between RSV A and B This method could provesignificant during a respiratory outbreak for surveillance Insuch situations multiplex real-time PCR assay such as thecommercial ldquoSimplexa Flu AB amp RSV kitrdquo that differentiatesinfluenza A virus influenza B virus and RSV is very useful[56] Likewise recently a rapid and sensitive detection assayfor RSV and other respiratory viruses was developed byIdaho Technologies christened as FilmArray (Idaho Tech-nologies Salt Lake City UT) This is an automatic real-timemolecular station with capability of nucleic acid extractioninitial reverse transcription and multiplex PCR followedby singleplex second-stage PCR reactions for specific virusdetection [57]

24 Microarray Correlating gene expression signatures withdisease progression of the patientrsquos individual genetic profileis possible by comprehensive understanding and interpre-tation of the dynamics of biome-interactions in the lungsThis would result in more efficient therapy for respira-tory diseases via the concept of personalized medicinewherein the microarray finds its applications Microarraysare the miniaturized assay platforms with a high-densityarray of immobilized DNA or protein Hybridization ofsample biomolecules to corresponding DNA or protein onthe chip is detected which allows determination of a varietyof analytes present in the samples in a single experiment[60] Microarray has proved to be a robust and reliabletool to understand the echelon of genomics transcriptomicsand proteomics Implementation of microarray with themetagenomic approach serves for rapid virus identifica-tion Microarray based identification and characterizationof viruses in clinical diagnostics are possible by designingoligonucleotide probes using the sequence data available inthe public database This approach is capable of discoveringnovel viruses even though there are no conserved genes thatcan be targeted by sequencing In the case of respiratorydiseases it can be executed by collecting the bronchoalveolarlavage enriched by nuclease treatment followed by filtrationand extraction of total nucleic acids Further the nucleic acidsare amplified by a randompriming-basedmethod referred toas sequence independent single primer amplification givingnear-full-length reads of genomes of RNA or DNA viruseswhich could then be compared to known viruses and usedfor designing the oligomers for the microarray [61] Thussequencing coupled with microarray is a powerful highthroughput diagnostic tool

The utility of a metagenomics based strategy for broad-spectrum diagnostic assay using microarrays was demon-strated for screening viruses like Rhino virus Parainfluenzavirus Sendai virus Poliovirus Adenovirus and RSV fromclinical samples [62] A well-known example is ldquoVirochiprdquowhich is a pan-viral microarray designed to simultaneouslydetect all known viruses and has comparable or superior sen-sitivity and specificity to conventional diagnostics [63 64]Similarly an influenza microarray ldquoFluChip-55 microarrayrdquofor the rapid identification of influenza A virus subtypesH1N1 H3N2 andH5N1 was developed [65]The procedure issimple and follows few steps including RNA extraction fromclinical samples subsequently their reverse transcriptionsecond-strand cDNA synthesis and then PCR amplificationof randomly primed cDNA The hybridization of nucleicacids with probes is done by fluorescent dyes or by analternative method of electrochemical detection The lattermethod relies on a redox reaction to generate electricalcurrent on the array for measurement Incorporation of Cy3fluorescent dye and hybridization to the microarray [66]has been empowered by specific algorithms to match thediagnostic needs leading to the final and critical step ofscanning and analysis [67]

The genes involved in the pathways of neuroactive ligand-receptor interaction p53 signaling ubiquitinmediated prote-olysis Jak-STAT signaling cytokine-cytokine receptor inter-action hematopoietic cell lineage cell cycle apoptosis and


cancer were upregulated in RSV-infected BEAS-2B cells RSVinfection up-regulated 947 and 3047 genes at 4 h and 24 hrespectively and 124 genes were common at both instancesMoreover 1682 and 3771 genes were downregulated at 4 h and24 h respectively and only 192 genes were same Respiratorydisease biomarkers like ARG2 SCNN1G EPB41L4B CSF1PTEN TUBB1 and ESR2 were also detected RSV infectionsigns and symptoms render a partial consequence of hostpathogen interaction but the transcription profile is betterexemplified by microarray The transcription profiles of RSVinfectedmice lungs and lymphnodes showed gene expressionof antigen processing and inflammation The response ishigher in lungs on the day 1 after RSV infection than day 3The gene expression profile shortly after RSV infection can beaccounted as a biomarker and can be scaled up for diagnosticswith in vitro and in vivo profiles [68 69]

Sometimes DNA microarrays do not confer their utilityin very specific investigations in the case of personalizedmedicine wherein the manifestation of disease occurs at thetranscription level This situation may arise in complicationsof RSV with other associated disorders like asthma [7071] As protein is the abundant functional biomoleculereflecting the physiological or pathological state of the organprotein microarray profile is an option to access underthis situation [72] Protein microarray has analytical andfunctional applications to study the protein-protein protein-DNA and protein-ligand interactions These features enablethe profiling of immune responses and are thus importantfor diagnostics and biomarker discovery Protein microarraycan serve as a rapid sensitive and simple tool for large-scaleidentification of viral-specific antibodies in sera [73 74] Anextensive study was done during the 2002 SARS pandemicusing coronavirus protein microarray to screen antibodies inhuman sera (gt600 samples) with gt90 accuracy and at leastas sensitive as and more specific than the available ELISAtests [73]Thus this systemhas enormous potential to be usedas an epidemiological tool to screen viral infections

25 Mass Spectroscopy Mass spectroscopy (MS) has becomea method of choice for molecular investigation of pathogensas the reliability is reinforced due the well-characterizedsequence information of nucleic acids or proteins and evenfor intact viruses [86 87] But pragmatic usage of MS ispossible when coupled with various chromatography andaffinity-based techniques The combination of affinity basedviral detention and nucleic acid basedMS serves as a solutionfor low detection limits Affinity-based methods employingnanotechnology can be used to find traces of target pathogento improve detection limits PCR amplification of pathogennucleic acid combined with MS can be used as substitutes[88ndash90] MS has the advantage of rapid identification of mul-tiple viruses at the same time and even identifies the proteinmodification status [86 91] Now MS is no longer confinedto proteomics based analysis and MS based genomics havebecome common practice There are several variations ofMS like Matrix-assisted laser desorptionionization massspectroscopy (MALDI-MS) Surface-enhanced laser desorp-tionionization mass spectroscopy (SELDI-MS) Bioaerosolmass spectrometry (BAMS) Pyrolysis gas chromatography

mass spectrometry (PyGCMS) Capillary electrophoresis-MS and Liquid chromatography mass spectrometry (LC-MS) are the few examples that are used for the identificationof pathogens [89] But the MS technique that promisesrobust practical application for pathogen detection is theelectrospray ionization mass spectrometry (ESI-MS) [92 93]Figure 3(a)

A notable ESI-MS for global surveillance of influenzavirus was accomplished by Sampath et al [75] by couplingMSwith reverse-transcription PCR (RT-PCR)The study cor-rectly identified 92 mammalian and avian influenza isolates(which represented 30 different H and N types including 29avian H5N1 isolates) The analysis showed more than 97sensitivity and specificity in the identification of 656 clinicalhuman respiratory specimens collected over a seven-yearperiod (1999ndash2006) at species and subtypes levelThe surveil-lance of samples from 2005-2006 influenza virus incidenceshowed evidence of new genotypes of the H3N2 strains Thestudy also suggested approximately 1 mixed viral quasi-species in the 2005-2006 samples providing insight into viralevolution This study led to a number of RT-PCRESI-MSbased detection methods for RSV and related respiratoryviruses like influenza A and B parainfluenza types 1ndash4adenoviridae types AndashF coronaviridae human bocavirus andhuman metapneumovirus screening [76] These assays had879 accuracy compared to conventional clinical virologyassays and pathogens undetected by traditional clinical virol-ogy methods could be successfully detected The advantageof RT-PCRESI-MS platform is the ability to determine thequantity of pathogens detailed pathogen characterizationand the detection of multiple pathogens with high sensitivityand efficiency

26 Nanotechnology Based Detection Recent advancementsin nanotechnology have changed the perception and perspec-tive of research When scaled down to nanometers the prop-erties of matter change and nanotechnology exploits thesenew properties and harnesses them with the existing tech-nologies to exceptional capabilities Techniques discussedbelow are examples of the nanotechnology based detectionapproaches which utilize basic traditional but indispensabledetection techniques

261 Nanoparticle Amplified Immuno-PCR Immuno-PCRa combination of ELISA and PCR is used widely for thedetection of various bacterial and viral antigens with lowertiters as meagre as zepto moles [58] Perez et al [59] reporteda modification of immuno-PCR using gold nanoparticlesfor RSV detection Target extraction was enhanced by usingmagnetic microparticles (MMPs) functionalized with anti-RSV antibody to capture the antigen (RSV) The MMP-RSV complex is then countered with gold nanoparticlesfunctionalizedwith both palivizumab (Synagis) an anti-RSVF protein antibody and DNA sequence partially hybridizedwith a tag DNA sequence (fAuNP) The MMP-RSV-fAuNPcomplex is then heated to release the partially hybridizedtag DNA sequence which is then quantified from thesupernatant by real-time PCR These modifications enableddetection of RSV even at 41 PFUmL This assay offers


ESI-MS Identification of virus

Virus A Virus B Virus C Virus D

PCRamplicon

(a) PCR-Electrospray IonizationMass Spectroscopy

Silver filmSilver nanorods

Silver vaporsource

Oblique vapor desposition

Spectra


Laser

Silver nanorods

(b) Surface Enhanced Raman Spectroscopy

Figure 3 A schematic representation of biophysical method of RSV detection (a) PCR-electrospray ionization mass spectroscopy and (b)Surface enhanced Raman spectroscopy

a 4000-fold improvement in the limit of detection over ELISAand a 4-fold improvement over detection when comparedwith real-time RT-PCR [59]

262 Live Cell RNA Imaging Live cell imaging can pro-vide the benefit of effective diagnosis and treatment ifthe capability of identifying monitoring and quantifyingbiomolecules is developed However development of suchsystems for detection of viral agents is difficult especially inthe early stages of infection With the advent of molecularbeacon technology it is now possible to track mRNA ofhost and RSV [32] Based on this framework a modifica-tion by oligonucleotide-functionalized gold nanoparticulateprobe was suggested as an improvement wherein a goldnanoparticle is functionalized withDNAhairpin structure bythiol linkage The DNA is so designed that the loop portionhas a complimentary sequence to RNA to be detected andthe 51015840 stem is linked to gold nanoparticle by thiol group andthe 31015840 end is linked to a fluorophore On hybridization to thetarget RNA with loop the fluorophore goes away from thequenching gold and the emission is tracked Thus live imag-ing of mRNA is possible This mechanism of hairpin DNAfunctionalized gold nanoparticles (hAuNP) was executed byJayagopal et al in detecting RSV mRNA in HEp-2 cells [33]This technique offers the advantage of real-time detectionof multiple mRNA at the same time including the mRNAof RSV and glyceraldehyde 3-phosphate dehydrogenase ofthe HEp-2 cell Quantitative assay using this approach was

possible using DNA hairpin structures functionalized toa gold filament which is immersed in a capillary tubecontaining viral RNA and scanned for fluorescence The set-up was able to detect as low as 119 PFUs which was sim200times better than a standard comparison ELISA

263 Quantum Dots Immunofluorescence microscopybased detection of RSV that is direct fluorescent-antibodyassay (DFA) is considered as gold standard [39] but thecomparative efficacy of DFA with other assays does not seemto have reached a consensus [39 40 94ndash97] The primepossible disadvantages and inconsistencies of DFA can beattributed to the fading of the dyes conjugating antibodieswith dyes limited sensitivity due to background staining andexcitation at two different wavelengths [26ndash28] To addressthese issues fluorescent nanoparticles that is quantumdots (QDs) appear as promising candidates for field clinicaldiagnostics Due to their inorganic nature they are lesssusceptible to metabolic degradation QDs are photostablethat is they do not lose fluorescence on long exposure to lightand can be excited at the same wavelength while radiating atdifferent wavelengths and hence can be used for multiplexing[27] Various successful attempts were made to ameliorateRSV detection using QDs The progression of RSV infectionin the HEp-2 cell line was studied using confocal microscopyby QDs probing F and G proteins and it was found thatthis method was more sensitive than real-time quantitativeRT-PCR particularly at early infection [29] This approach


was used by Tripp et al in vitro on Vero cell lines andwas extrapolated by an in vivo BALBc mice study whichconcluded the approach beyond diagnostics as it can be usedfor multiplexed virus andor host cell antigen detection andintracellular tracking studies [28]

Flow cytometry is nowwidely used in diagnostics and thereliability is improved because millions of cells are analyzedat a time and comprehensive data is produced [30] Trackingand targeting cellular proteins and the ease of multipleparameters correlation allow flow cytometry to be used forvarious qualitative and quantitative assays Flow cytometercould detect RSV with sensitivity and reproducibility [31]Agrawal et al [26] showed that antibody conjugated QDscould detect RSV rapidly and sensitively using the principlesof microcapillary flow cytometry (integrated with a fixed-point confocal microscope) and single-molecule detectionA 40-nm carboxylate-modified fluorescent nanoparticles andstreptavidin-coated QDs were used in their study whichcould estimate relative levels of surface protein expression

264 Gold Nanoparticle Facilitated Microarray The FDAapproved microarray systems semiautomated respiratoryvirus nucleic acid test (VRNAT) and the fully automated res-piratory virus nucleic acid test SP (RVNATSP) (NanosphereNorthbrook IL) are examples of the microarray based detec-tion systems for influenza A virus influenza B virus RSVA and RSV B [56 98] These systems are based on the effi-cient detection of microarray based hybridization using goldnanoparticlesThe hybridization between the oligonucleotideprobes and target DNARNA is detected specifically byhybridizing them again to gold nanoparticles functionalizedwith oligonucleotide and the signal is generated by goldfacilitated reduction of silver in the presence of a reducingagent [99]

265 Surface Enhanced Raman Spectroscopy Raman spec-troscopy works on the basis of the inelastic scattering ofmonochromatic radiation like near infrared visible or nearultraviolet interacting with an analyte with low-frequencyvibrational andor rotational energy But the signal generatedis low and hence the signal is enhanced using silver or goldmatrix substrates There are many modifications of Ramanspectroscopy but the most widely used one is the surfaceenhanced Raman spectroscopy (SERS) (Figure 3(b)) [77]Direct intrinsic indirect intrinsic and extrinsic detection arethree major SERS detection configurations [78] In contrastto IR spectroscopy the Raman spectra can be obtainedwithout the interference of water molecules and thus bio-logical analytes can be studied in their native conformationSERS is routinely used for various bioanalytical purposesdue to the rapid and nondestructive detection of analyteswith sensitivity specificity and precision even for a singlemolecule or live cells [79 80] SERS can be targeted tovarious analytes that may constitute DNA RNA proteinsor other organic compounds Nucleic acids are the preferredcandidates in biological SERS investigations as the influenceof base composition and sequence conformation (localandor global) of nucleic acids or intermolecular dynamicswith protein or ligand is correspondingly expressed as typical

spectral signature [81] Bacteria and viruses from variousbiological samples can be identified characterized and clas-sified from clinical samples [77 82] SERS can distinguishbetween DNA or RNA viruses like Adenovirus RhinovirusRotavirus [83] and RSV [84] Rotavirus is the common causeof gastroenteritis in children and the rapid and sensitivedetection was demonstrated with SERS [83] SERS is anestablished powerful tool for sensitive expedited and specificdetection of various respiratory pathogens like Mycoplasmapneumoniae and RSV Using silver nanorod arrays (NA)platform for SERS differentiation of M pneumoniae inculture and in spiked and true clinical throat swab sampleswas achieved [82] The notable sensitivity of SERS to resolvestrain level differences for RSV strains ALong A2 ΔG andB1 can be exploited for clinical diagnosis instrumentation[84]

3 Prevention

Prevention is the most important aspect of healthcare andhas prime contribution to the culmination of the diseasethan the treatment measures Effective preventive measuresreduce the mortality and economic burden of the diseaseUntil now there is no effective vaccine for prevention for RSVDirect or indirect contact with the nasopharyngeal secretionsor droplets (sneezing coughing and kissing) fomites andfood from infected patients can potentially transmit RSV Livevirus can survive on surfaces for several hours [100] but at thecellular level the viral spread is a series of systematic eventsof invasion including viral attachment and fusion followedby viral replication and protein synthesis The F proteinaccumulates in the host membrane and then surrounds thebudding progeny viruses thus spreading the infection toadjacent cells and exacerbating the infection (Figure 4)Thusthese proteins are considered as the potential candidate forthe development of prevention measures such as antibodiesDNA vaccines and subunit vaccines

31 Antibodies RSV has three envelope proteins F G andSH Both F and G are glycosylated and represent the targetsof neutralizing antibodies The RSV F protein emerged asa good vaccine candidate due to its conserved and vitalrole in cell attachment Passive immunization is a directapproach to counter RSV (Figure 5) Initially polyclonalantibodies from healthy human individuals resistant to RSVwere successful in preventing RSV infection in high riskinfants and these pooled and purified immunoglobins werepopular as RespiGam The monoclonal antibody specificallyneutralizing F protein conferred effective protection againstRSV as compared to RespiGam and this licensedmonoclonalantibody palivizumab (Synagis) is now used to passivelyprotect high risk infants from severe disease thus replacingthe RespiGam The efficacy of the recombinant monoclonalantibody palivizumab has been tested for prophylaxis andtherapy in immunocompromised cotton rats [101] Repeateddoses of palivizumab were required to prevent reboundRSV replication Palivizumab is administered alone or incombination with aerosolized ribavirin Palivizumab cannot


Antigenome

Transcription Replication

Protein synthesis

AssemblySH protein

N protein

Nucleus

P protein

L protein

Endoplasmicreticulum

Golgi complex

Life cycle of RSVRSV

Merging with the cell membrane

Budding RSVready to infectadjacent cell

F protein

G protein

Figure 4 A schematic representation of RSV life cycle

Modes of RSV inhibitionF protein G protein

+

+

+

+

+

Gold nanoparticles

Antibodies

No bindingto the cell



RSV

RSV

RSV

RSV

Drug

PeptideFusion inhibitors

No treatmentFusion Infection

No infection

No infection

No infection

Binding

Cell

Figure 5 A schematic representation of various compounds inhibiting RSV binding to the cell

cure or treat serious RSV disease but neutralization of RSVcan help in preventing serious RSV infections Motavizumabhas been found to neutralize RSV by binding the RSVfusion protein F after attachment to the host but before theviral transcription [102] Viral entry was not inhibited by

palivizumab or motavizumab when pretreated with RSV butthere was a reduction in viral transcription thus inhibitingboth cell-cell and virus-cell fusion most likely by preventingthe conformational changes in the F protein needed for viralfusion


The effective use of palivizumab is limited due to the costand its use in infants with high risk of bronchiolitis based onthe coverage by different healthcare systems [103] In spiteof these restrictions on palivizumab it has a wide societalimpact on use in infants with chronic lung disease due topremature birth or those with haemodynamically significantcardiac disease According to themodified recommendationsof the Committee on Infectious Diseases of the Centers forDisease Control and Prevention of RSV palivizumab is rec-ommended for infants with congenital heart disease (CHD)chronic lung disease (CLD) and birth before 32 weeks [104]Minimum 5 doses are recommended irrespective of themonth of the first dose for all geographical locations forinfants with a gestational age of 32 weeks 0 days to 32 weeks6 days without hemodynamically significant CHD or CLDThe new recommendations of the committee were aimed atthe high risk groups including infants attending child careor one or more siblings or other children younger than 5years living with the child Also the infants were qualifiedfor receiving prophylaxis only until they reached 90 days ofage Palivizumab although effective is costly and thus is notbeneficial to the recipients especially during the periodswhenRSV is not circulating A cost effective means of producingRSV F neutralizing antibodies was experimented in phagesand plants Much success in this regard of palivizumabproduction was observed in theNicotiana benthamiana plantsystem which offered glycosylation and high productionat lower upstream and equivalent downstream cost whencompared tomammalian derived palivizumabThe efficacy ofthe plant derived palivizumabwasmore than themammalianderived palivizumab or the plant derived humanmonoclonalantibodies in cotton rats [105]

32 DNA Vaccines RSV genome codes structural and func-tional proteins that are immunogenic and referred for vaccinedevelopment against RSV DNA based vaccines were devel-oped based on these proteins because the concept was simpleas it involved aDNA fragment coding part orwhole protein ofRSV that was inserted into an appropriate expression plasmidvector under a constitutive promoter control (Figure 6)The initial work with this approach was successful in theexpression in cells and in vivo murine models to eliminatethe RSV infection but the problem of RSV associated Th2type immune response was persistent This problem wasattempted to be resolved by manipulating the parametersof choice of the protein to be expressed the expressionvector adjuvants formulations and intracellular stability ofthe plasmid

Mice challenged with the RSV-G construct had balancedsystemic and pulmonaryTh1Th2 cytokines andRSVneutral-izing antibody responses [106] However the RSV F proteingene is considered as a widely used and prospective targetfor the development of vaccines and is often a favouritemodel for DNA vaccines against RSV [107] But the wildtype RSV F protein expressed from DNA plasmid waspoorly expressed so a codon optimized DNA vaccine wasdesigned for better in vivo expression and hence was moreimmunogenic and reduced the RSV titer [108] To addressthe low immunogenicity of F protein the antagonistic activity

of RSV for IFN and immunopathology a construct wasdesigned by inserting the F gene into Newcastle diseasevirus (NDV) vector (NDV-F) This modification served thepurpose of higher elicitation of IFN by NDV-F than RSVprotection against RSV infection without immunopathologyand enhanced adaptive immunity in BALBc mice [109] Wuet al [110] developed a DNA vaccination strategy againstRSV using a mucosal adjuvant Two DNA vaccine vectorsnamely DRF-412 and DRF-412-P containing residues 412ndash524 of the RSV F gene were cloned into the phCMV1DNA vaccine vector The DNA vaccine vectors DRF-412contained the cholera toxin gene region called ctxA2B actingas a mucosal adjuvant The DNA vaccine was successfullyexpressed in mouse muscle tissue which was confirmed byimmunohistological analysis and RT-PCR The immunizedmice induced neutralization antibody systemic Ab (IgGIgG1 IgG2a and IgG2b) responses and mucosal antibodyresponses (IgA)whichmimicked the challengewith live RSVThemice immunized with the DRF-412 vector contained lessRSV RNA in lung tissue and induced a higher mixedTh1Th2cytokine immune response and had better protection thanthose immunized with the DRF-412-P vector which wasconfirmed by lung immunohistology studies [110] A rationalapproach to confer protection against various pathogensthrough single vaccination ormore often known as combinedor composite vaccines was employed for RSV influenza Avirus (INF-A) and herpes simplex virus type-1 [111] Micewere immunized either by injection or by gene gun (goldbeads as carrier of the plasmid DNA) with mixture of fourplasmids INF-A haemagglutinin (HA) INF-A nucleoprotein(NP) HSV-1 glycoprotein D (gD) and RSV glycoprotein FThis led to protection of mice from the respective pathogensof challenge in addition it also offered protection fromMycoplasma pulmonis challenge as well [111] Recently severaldevelopments have been made in the field of recombinantvaccines In one such approachMycobacterium bovisBacillusCalmette-Guersquorin (BCG) vaccine was modified to carry RSVN or M2 and was tested and found to establish the Th1 typeimmunity in RSV challenged mice [112] The recombinantvaccine also elicited the activation of RSV specific T cellsproducing IFN-120574 and IL-2 along with reduction in weightloss and lung viral protein load thus establishing a Th1-polarized immune response

Bactofection is bacteria mediated transfer of plasmidDNA into mammalian cells Xie et al employed this interest-ingway of delivering and expressing theDNAvaccine inmiceagainst RSV [113] This approach serves the purpose of natu-rally activating the immunostimulatory response of the hostand also delivery of the DNA vaccine Attenuated Salmonellatyphimurium strain SL7207 expressing vector pcDNA31Fcontaining theRSVF genewas orally administered to BALBcmice triggering efficient antigen-specific humoral cellularand mucosal immunity [113 114]

A novel method of developing RSV DNA vaccine wasdevised by replacing the structural genes with RSV genesin an attenuated strain of Venezuelan equine encephalitisvirus (VEEV) VEEV has an ssRNA (+) genome and con-tains a strong subgenomic promoter The replicon particleswere prepared by providing helper RNAs encoding VEEV


Naked DNAvector

Vaccination

RSV

DNA functionalizednanoparticles

RSV

RSV challenge

RSV challenge

DNA encapsulatednanoparticles

Mice protected

Expression

DNA vaccine

Viral DNA

Vector

vector

No vaccination RSV infection

Figure 6 A schematic representation of a simple DNA vaccine administered as a naked DNA vector or functionalized nanoparticle or as anencapsulation for controlled delivery

capsid and envelope glycoprotein which comprise structuralproteins and all of these when transfected into Vero cellsresulted into a replicon particle that could independentlysynthesize the RSV protein thereby activating the immuneresponse and protection The system could be modulated bythe administration of helper RNAsThis strategy was appliedto themice and rhesusmonkeymodels conferring protectionagainst RSV and a desirable extent of a balancedTh1Th2 typeimmune response was received [115]

33 Subunit Vaccines Several approaches have been con-sidered for developing an effective vaccine against RSVincluding the formalin inactivated RSV vaccine developed inlate 1960s which instead resulted in an enhanced infection[116] An efficient RSV vaccine would be the one with properbalance between immunogenicity and protectionwithout anyallergic response [8]

RSV F has widely been accepted as the vaccine candidatedue to its conserved nature among various strains as well asamong the other paramyxoviruses [8 117ndash119] An antigenicregion corresponding to RSV F protein (region 255ndash278)was cloned into a vector with ctxA2 B gene of choleratoxin and named as rF255 which elicited a helper T celltype 1 immune response in mice It also resulted in higherexpression of serum neutralizing antibody in immunizedmice [120] Similarly a multivalent recombinant protein wasdeveloped by cloning RSV F M2 and G protein into abacterial pET32a (+) vector (called rFM2G) which resultedin enhanced serum IgG titers [121] rFM2G was also usedin conjunction with flagellin as an adjuvant which did notincrease the IgG titers In an effort to address the issues ofimmunogenicity several studies have used the approach ofcombining other viruseswithRSVgenesOne such study uses

virus like particles (VLP) developed fromNP andM proteinsof Newcastle disease virus (NDV) and a chimeric proteinconsisting of cytoplasmic and transmembrane domains ofNDV HN protein and ectodomain of the RSV G protein(HG) [122] These VLPs resulted in an immune responsebetter than UV-inactivated RSV and provided completeprotection in mice from RSV even at a single dose withelicited neutralizing antibodies The VLP-HG-immunizedmice did not show enhanced pathology as compared toFI-RSV In another approach to solve the immunogenicityrelated problems in devising an RSV vaccine a recombinantvector based on either murine PIV type 1 or Sendai virus wasused to deliver RSV G protein through reverse genetics [123]This provided effective protection against RSV in cottonrats Another similar study using Sendai virus (SeV) as avaccine carrying RSVF gene provided protection against fourpathogens including hPIV-1 mouse PIV-1 hPIV-3 and RSV[124] This approach has been used to compare two vaccinemodels one with SeV backbone and the other with PIV-3backbone The SeV based vaccine showed a decrease in RSVload in African green monkeys lung titers

Another study with recombinant vaccine utilizes recom-binant simian Varicella viruses (rSVVs) which express RSVG and M2 protein genes and was evaluated in the Verocell line [125] Such a recombinant vaccine approach couldbe very useful in elderly people with risk of infection withVaricella and RSV as well as children with chickenpox andRSV rSVV based vaccines resulted in enhanced immuneresponses to RSV antigens serving as suitable vaccines inrhesus monkeys A very similar approach was adopted fordeveloping a recombinant alphavirus or immune stimu-lating complex (ISCOM) antigen against RSV [126] Therecombinant vector was designed by using a self-abortive


alphavirus called Semiliki forest virus carrying RSV F andG genes The advantages of using such recombinant abortiveviruses are that they can undergo viral infection just onceeliminating chances of adverse effects resulting from furtherinfection and infecting the host cell resulting in expressionof the inserted RSV gene followed by humoral and cellmediated immune responseThe ISCOMusedwas composedofQuillaja saponin lipids andRSV antigens having adjuvantproperties as well ISCOM has been shown to enhanceantibody production T cell proliferation and MHC class-I responses as well as RSV specific neutralizing antibodiesIgG and IgA The recombinant vaccine SFVF or SFVFGresulted in IFN gamma response along with resistance toRSV infection without worsened RSV disease whereas theresults were opposite in ISCOMFGwith enhanced goblet cellhyperplasia post-RSV challenge

34 Nanovaccines Recently several applications of nan-otechnology have appeared in the development of vaccinespopularly known as Nanovaccines DNA vaccine is prone torapid degradation when introduced into an animal systemso to increase the retention and increase the efficacy of theDNA vaccines they can be encapsulated into a polymerthat will protect and facilitate controlled release (Figure 5)Various synthetic or natural polymers are now experimentedfor targeted delivery and controlled release of the carrierChitosan is the polymer of great interest in respiratorydisease treatment because of its mucoadhesive property andbiodegradability which balances the purpose of longer reten-tion and controlled release of carrier molecules encapsulatedThus chitosan nanoparticles are being developed againstRSV A DNA vaccine (DR-FM2G) constitutive expressionvector consisting of antigenic regions of RSV F M2 andG genes driven by human cytomegalovirus promoter wasencapsulated using chitosan nanoparticles (DCNPs) Theadvantages of DCNPs are that it was more stable than nakedDNA or chitosan and so ideal for protection against DNAdegradation by nucleasesThe sustainability of the DCNPs inmice was more than naked DNA which was correspondinglyindicated by higher RSV protein expression evident byimmunohistochemical and real-time PCR studies [127 128]A similar study was conducted by Eroglu et al [129] wherethe highly conserved RSV F gene was cloned into pHCMV1expression vector and encapsulated into poly-hydroxyethyl-methacrylate nanospheres coated by chitosan and transfectedinto Cos-7 cells The transfection efficiency of the systemwas at par with commercially available transfecting agents Invivo studies with the BALBC mice also indicated F proteinexpression and reduced RSV infection

A vaccine based on the recombinant RSV N gene ringscalled N-SRS enclosing a bacterial RNA has been developedand assessed intranasally in BALBc mice [130] N-SRS wasadjuvanted with E coli enterotoxin LT (R192G) with efficientprotection against RSV and high titers of IgG1 IgG2a andIgA and so forth Although this nanovaccine elicited a mildinflammatory response in airways of the mice it enhancedthe expression of antigen specific CD4+ and CD8+ responsesIn another novel approach nanoemulsion (NE) was usedas an adjuvant to mucosal RSV vaccine in a mouse model

and found to provide significant protection against RSValong with RSV specific humoral responses and an enhancedTh1Th2 response [131] The NE rendered the RSV ineffectivewhen treated for 2 or 3 hours with a reduction in viral titerup to 10-fold as compared tomedia controls NE-RSV also ledto higher expression of RSV specific antibody with significantdecrease in viral load no hyperreactivity andnoTh2cytokineinductionThus this novel approach seems to be a promisingand safe vaccine option against RSV

The bitter episode of formalin inactivated RSV vaccinehas impeded the vaccine development and in fact has raisedserious concern in the use of native RSV or its componentsThis approach was revived with a novelty wherein theengineered RSV F protein aggregates formed nanoparticlesand were used as vaccine and these nanoparticles inducedprotective immunity in cotton rats [132] To combat RSV hostneutralizing antibodies are always more preferred than thetherapeutic antibodies But the epitopes of the neutralizingantibodies are larger than those of the therapeutic neu-tralizing antibodies (palivizumab and motavizumab) Thusthe use of these epitopes for neutralizing antibodies as avaccine requires the retention of immuno functional con-formation while getting rid of the undesired protein RSV Foligomeric protein nanoparticle was synthesized by insertingrecombinant RSV F gene into Baculovirus and expressedin Sf9 insect cell lines This resulted in high recombinantprotein expression compared to native protein This alsoresulted in the conformation of rosette nanoparticles whichwas the aggregate of multiple RSV F oligomers found to beimmunogenic The study was extrapolated to phase 1 humanclinical trials which showed its safety and efficacy againstRSV [133]

35 Nutrition Nutrition is not considered as treatmentbut is a prerequisite for homeostasis and any nutritionalimbalance attracts disorders and diseases Supplementarynutrients could reverse the adverse effects of the disordersand diseases so in sensu nutrition can serve both as treatmentand prevention Carbohydrate rich diets and diet lackingantioxidants like fruits and vegetables during pregnancy aresuspected for RSV susceptibility [134] Resveratrol (trans-345-trihydroxystilbene) a polyphenol from grapes hasdemonstrated reduction in RSV replication and inflamma-tion [135 136] A study demonstrated that the cord blooddeficient in vitamin D was associated with RSV bronchiolitisthe neonates were at higher risk of RSV in the first yearof life [137] Vitamin D inhibits NF-120581B signalling whichis responsible for RSV inflammation without affecting theantiviral activity of the host [138] Lower levels of micronutri-ents like zinc copper selenium and retinol (Vitamin A) andalpha-tocopherol (Vitamin E) were observed in the childrenaffected by RSV and human metapneumovirus [139] In onestudy RSV infected children were administered vitamin Ato compensate the lower vitamin A serum level which wasfound beneficial for children with severe RSV infection [140]Also probiotic diet has shown to boost resistance againstpathogen by modulating immune response as in case ofRSV Lactobacillus rhamnosus (a probiotic bacteria) treatedBALBc mice showed significantly reduced lung viral loads


and pathology after the RSV challenge [141] These studiesindicate the importance and association of nutrition withRSV susceptibility

4 Treatment

There are very limited treatment options available for RSVHowever there are many drugs for the symptoms associatedwith RSV infection The target genes and proteins vital forRSV infection (discussed in Section 3) are important fordeveloping preventative and treatment measures The modeof action and potency of a drug determines the approachof prophylactic or curative application Considering theproposed life cycle of RSV theoretically there are numerousmodes to interfere with RSV infection but these options maynot be practical Replication transcription and fusion are thefew target processes for drug development against RSV Afocus is therefore on development of potent drug which holdsconformity in the human trials Some of the approaches aredescribed below that promise to be a potential treatment forRSV (Table 2)

41 Antiviral Drugs Ribavirin or 1-[(2R 3R 4S 5R)-34-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-1H-124-tri-azole-3-carboxamide is a widely used broad spectrumsynthetic anti-viral drug for both DNA and RNA virustreatment Oral and nasal administration of ribavirinfor treatment of severe lower respiratory tract infectionscaused by RSV and influenza virus are an option [104]Ribavirin is phosphorylated in the cells and competes withadenosine-51015840-triphosphate and guanosine-51015840-triphosphatefor viral RNA-dependent RNA polymerases in RNAviruses However the mechanism of ribavirin differs forDNA viruses as it is a competitive inhibitor of inosinemonophosphate dehydrogenase (IMPDH) causing deletionof GTP and messenger RNA (mRNA) guanylyl transferase(mRNA capping enzyme) and adversely affecting proteinsynthesis [142 143] The exact mechanism of anti-viralactivity of ribavirin against RNA and DNA viruses is stillnot clear The usefulness of ribavirin against viruses isnot only due to its anti-viral activity but also due to itscapability to modulate the immune system Ribavirin issuggested to have immuno-stimulatory effects on Th cells[144] The derivatives of ribavirin such as viramidinemerimepodib and other IMPDH inhibitory moleculeslike mycophenolate and mizoribine have shown antiviralactivity against the hepatitis C virus and hence there isscope for investigating them as potential anti-RSV drugs[143 145] There are many other compounds that can inhibitRSV replication and a well-known compound RSV 604((S)-1-(2-fluorophenyl)-3-(2-oxo-5-phenyl-23-dihydro-1H-benzo[e][14]diazepin-3-yl)-urea) showed promising resultsagainst RSV [146]

A derivative of antibiotic geldanamycin 17-ally-lamino-17-demethoxygeldanamycin (17AAG) and 17-dimethylam-inoethylamino-17-demethoxygeldanamycin (17DMAG) tar-geted against cancer has now attracted researchers due to itsantiviral property These compounds are HSP90 inhibitorsand thus helpful against RSV as RSV is dependent on heat

shock protein (HSP90) for its replication [165] The anti-RSV activity of these compounds was seen in human airwayepithelial cells (HAEC) and is considered as drug resistanttherapeutics due to the highly conserved target chaperonprotein These compounds are also known to inhibit repli-cation of HPIV influenza virus and rhinovirus The anti-viral activity dose is not toxic to the cells and inhalationmode of treatment can increase the local efficacy and avoidunnecessary exposure to other organs [166]

42 Fusion Inhibitors Recent advances in the developmentof anti-viral drugs include the fusion inhibitors The fusioninhibitors are usually synthetic compounds or moleculesinterrupting the fusion of virus with the host cell usually bybinding the fusion proteins (Figure 5) The fusion inhibitorshave beenwidely studied as anti-viral agents in several virusesincluding HIV RSV Henipavirus Hendra virus Nipah virusParamyxovirus metapneumoviruses HIV and RSV [11 167ndash173] The first reports of the use of peptide(s) as fusioninhibitors include the development of DP-178 a syntheticpeptide based on the leucine zipper region of the HIV fusionglycoprotein gp41 [167] which showed an IC50 at 038 nMagainst HIV-1 Fusion inhibitors for the paramyxoviruseshave also been developed based on the conserved regionof the fusion protein F The F protein is widely known forits conserved nature among the Paramyxoviridae family [7]Lambert et al [174] developed the fusion inhibitors belongingto the conserved heptad repeat (HR) domains of F1 regionof F protein which is analogous to the peptides DP-107 andDP-178 of HIV gp41 These fusion inhibitors were testedagainst RSV human parainfluenza virus 3 and measles viruswhich showed antiviral activity specific to the species oforigin DP-178 is an FDA approved anti-HIV drug withInternational Nonproprietary Name (INN) Enfuvirtide andtrade name Fuzeon Out of the peptides tested the peptideT-118 developed from RSV was the most effective with anEC50 of 0050120583MThese fusion inhibitors were then furthercharacterized and tested It was shown that different fusioninhibitors derived from same HR region differ in their anti-viral activity [147]TheHR121 andHR212 peptides showed anIC50 of 33 and 795 120583M respectively against RSV Similarlyanother peptide inhibitor was developed from Rho-A whichshowed inhibition of syncytia formation induced by RSV[148] RhoA (a small GTPase) is involved in many biologicalprocesses and was shown to bind the RSV-F protein at aminoacids 146ndash155 A peptide derived from the RSV F bindingdomain of RhoA (RhoA77-95) was shown to inhibit RSV andPIV-3 infection and syncytium formation block cell-to-cellfusion and reduce viral titers and illness in mice

The fusion inhibitors are not only limited to peptideinhibitors but a range of chemical inhibitors have also beentested against RSV and benzimidazoles are well-knownfusion inhibitors [175] A lead compound was identifiedto subsequently synthesize an analogue JNJ2408068 a lowmolecular weight benzimidazole which showed high anti-viral activity It had an EC50 of 016 nM 100000 times betterthan that of ribavirin This compound showed anti-fusionactivity against RSV in a dual mode of action includingprevention of cell-virus fusion activity as well as cell-cell


Table2Com

paris

onof

different

treatmentapp

roachesfor

RSV

Treatm

ent

Mechanism

Exam

ple

Remark

References

Antivira

ldrugs

Replicationinhibitio

nRS

V60

4Eff

ectiv

eagainstRS

Vbu

tadverse

effecto

ntheh

ost

[104]

[142ndash146]

Mutation

Ribavirin

vira

midinemerim

epod

ibInhibitoro

fino

sinem

onop

hosphate

dehydrogenase

Ribavirin

mycop

heno

latem

izoribine

Immun

ostim

ulatoryeffects

Ribavirin

Fusio

ninhibitors

Inhibitin

gfusio

nproteinattachmenttocell

Peptidemdash

HR121H

R212R

hoA

Chem

icalmdashBM

S-433771R

FI-641

Peptidefusioninhibitorsprom

ising

anti

RSVdrugchemicalfusio

ninhibitorshave

sidee

ffects

[147ndash150]

Nanop

articles

Inhibitin

gattachmenttocell

Silver

nano

particlesgold

nano

particles

Emerging

fieldcon

clusiv

estudies

requ

ired

[151152]

Antise

nsetherapy

RNAinterfe

rence

siRNA-ALN

-RSV

01Ph

osph

orod

iamidatem

orph

olinooligom

ersEff

ectiv

eand

safeA

LN-RSV

01completed

phaseIIb

clinicaltrails

[153ndash157]

Ethn

obotanicals

Prob

ablyfusio

ninhibitors

anti-inflammatory

Plantextractsmdash

Cinn

amom

umcassia

Cimicifuga

foetida

Sheng-Ma-Ge-Gen-TangGingeretc

Decoctio

ns-m

odified

Dingchu

an

Liu-He-Tang

water

extracto

fLicorice

Prom

ising

butcon

clusiv

estudies

requ

ired

[158ndash164

]


fusion activityHowever it was found ineffective against otherviruses of the family including HPIV-3 and measles virusA vast screening of as many as 16671 compounds (sourceChemBioNet library) was conducted for anti-RSV activityin vitro and two novel compounds N-(2-hydroxyethyl)-4-methoxy-N-methyl-3-(6-methyl[124]triazolo[34-a]phthal-azin-3-yl)benzenesulfonamide (named as P13) and the14-bis(3-methyl-4-pyridinyl)-14-diazepane (named as C15)were mined which reduced the virus infectivity with IC50values of 011 and 013 120583M respectively [176]

Recently several groups have reported synthetic fusioninhibitors of RSV especially the benzotriazole derivatives[177] After evaluating the structure-activity relationship(SAR) of these compounds named as series 1 compoundsit was observed that the topology of the side chains of thesecompounds is important and facilitates the modificationof their physical properties as many of these compoundsshowed poor therapeutic indices (cytotoxic effects) to thehost cells tested In order to address these issues a secondseries of derivatives of the compound 1 were developed andevaluated for SAR and their functionality as fusion inhibitors[178] These compounds were developed from 1 which hada tolerant diethylaminoethyl side chain with both polar andnonpolar functionality against RSV and had a replacementof the benzotriazole with benzimidazole-2-one These werepotent inhibitors of RSV in vitro These compounds werenamed 2 and had an additional structural vector absentin 1 which accounted for enhanced potency as fusioninhibitors and served as the base for further developmentof fusion inhibitors Further these group 2 compounds weremodified by introducing acidic and basic functional groupsinto the side chains [179] The oxadiazolone had anti-RSVactivity comparable to that of ribavirin whereas the estermodified group 2 compounds were suitable for oral admin-istration These studies further led to the identification ofa benzimidazole-2-one derivative called BMS-433771 whichwas an orally active RSV inhibitor [149] Another compoundstudied was the 5-aminomethyl analogue 10aa with potentanti-RSV activity towards BMS-433771 resistant RSV Thecompound BMS-433771 was further modified at side chainsand with the introduction of an aminomethyl substituentat the 5-position of the core benzimidazole moiety [180]The aminomethyl substitution in the benzimidazole ring wasfound to enhance the antiviral activity

Furthermore the consecutive modification of benzimi-dazole resulted in benzimidazole-isatin oximes which wereevaluated for anti-RSV activity [181]The compoundwas ana-lyzed for its antiviral activity cell permeability andmetabolicstability in human live microsomes Several other derivativeswith modification such as O-alkylation and addition ofnitrogen atoms to isatin phenyl ring were implemented alsoto enhance antiviral activity Three compounds 18j 18i and18n were shown to have anti-viral activity against RSV in theBALBc mice Further a compound RFI-641 was identifiedwhich was found to be the most potent anti-RSV agentinhibiting RSV both in vitro and in vivo and is in phase Iclinical trials RFI-641 is a biphenyl triazine synthesized bycoupling diaminobiphenyl to two chlorotriazine moleculesunder microwave conditions [150] RFI-641 was found to be

effective against 6 laboratory and 18 clinical viruses at con-centrations between 0008 and 011mM (0013ndash018mgmL)The compound reduced the viral load to an extent of 17 logsin the African green monkey model and also in mice andcotton rats In order to further enhance the antiviral activityof RFI-641 it was modified by replacing its triazine linkerswith pyrimidine [182] However this modification did nothave much difference in the anti-viral activity thus renderingthis modification not much of practical useThere are severalnovel nonbenzimidazole based compounds showing anti-RSV activity in vitro but a more polar compound thiazole-imidazole 13 was selected on the compound potency mod-erate permeability and low metabolic rate in rats and moredetailed in vivo studies are further anticipated [183]

43 Nanoparticles It has been established that metals likesilver [184] and gold [185] have anti-microbial activity butcytotoxic effects of these reactive metals make them unsuit-able for their use in humans The reactivity and behaviour ofmetals can be modulated by reducing their size to nanoscaleCarbon nanotubes (CNTs) are emerging nanomaterials forbiomedical application [186] Polyvinylpyrrolidone (PVP)conjugated silver nanoparticles showed low toxicity to HEp-2 cells at low concentrations and exhibited 44 RSV inhi-bition [151] (Figure 6) Singh et al used fusion inhibitorpeptide functionalized gold nanoparticles and carboxylatedgold nanoparticles of size 13 nm against RSV which showed83 and 88 inhibition of RSV respectively [152] Similarapproach was employed by recombinant RSV F proteinfunctionalized on gold nanorods [187] The emergence ofnanotechnology has opened new avenues for RSV treatment

44 Antisense Treatment RNA interference (RNAi) whichis a normal cellular event has become a powerful means ofcontrolling gene regulation The interference mediated bysiRNA was used against human immunodeficiency viruspoliovirus hepatitis C and parainfluenza virus (PIV) in cellculture [188ndash190] The concept of inhibiting RSV infectionusing targeted antisense mechanism was applied by Jairathet al by silencing the RSV-NS2 gene [191] Following theRNAi approach Bitko et al designed siRNA against the Pgene of RSV and PIVwhich protectedmice against individualand mixed infections upon intranasal administration [192]The effectiveness of siRNA action was observed with andwithout the use of transfection reagents This approach wasalso effective when targeting the RSV-F gene [193] Similarwork on HEp-2 cell lines was replicated using four siRNAdesigned to silence RSV F gene which showed inhibitoryaction against RSV at various concentrations [194] Silencingdifferent RSV genes too had an inhibitory action on theRSV a plasmid encoding siRNA which was complexed withchitosan targeting RSV-NS1 gene decreased RSV infectionin BALBc mice and Fischer 344 rats and also reducedthe associated inflammation [195 196] Zhang et al showedthat siRNA nanoparticle targeting RSV NS1 gene resulted inincreased IFN-120573 and IFN-inducible genes in A549 cells andin human dendritic cells elevated type-1 IFN and increaseddifferentiation of CD4+ T cells to Th1 cells [196] Also


mice treated with siNS1 nanoparticles exhibited significantdecrease in lung viral titers and inflammation

An interferon-inducible enzyme 2-5A-dependent RNasepresent in higher vertebrates requires 51015840-phosphorylated2101584051015840-linked oligoadenylate (2-5A) for its endoribonucleaseactivity against single-stranded RNAs This feature of 2-5A-dependent RNase is looked upon as an effective RSVtreatment [197] RSV upon infection elicits immune responseof the host and particularly the interferon levels [68] andthis phenomenon is exploited for the anti-RSV activity of2-5A-dependent RNaseL The endoribonuclease activity of2-5A-dependent RNaseL was used for targeting RSV M2gene specifically by covalent 21015840ndash51015840 oligoadenylate targetantisense which resulted in the reduction of RSV replicationThe endoribonuclease activity had negligible effect with aninactive dimeric formof 2-5A linked to antisense 2-5A linkedto a randomized sequence of nucleotides and antisensemolecules lacking 2-5A and did not affect the other RSVsor cellular RNAs [198] Additionally to widen the range ofthe approach the effects of modification of oligonucleotidesand RNA target sites were studied [199] This model wasimproved with respect to the specificity and activity bya chimera of 2-5A-antisense christened as NIH351 Thesequence information of RSV genome was used to developNIH351 and was 50- to 90-fold more potent against RSVstrain A2 than ribavirin [200] Administration of siRNA incombination with ribavirin was recommended for effectivetreatment [201] The parent molecule was chemically modi-fied to further increase the in vivo stability and specificity andpotency of NIH351 The resulting new version RBI034 wassim50 more effective than the parent molecule against RSV(strain A and B) and was not cytotoxic in the effective doseranges RBI034 treatment of African green monkeys showspromising results [202]

Alvarez and coworkers [153] came up with a new RSV-NS1 gene specific siRNA (ALN-RSV01) having a broadspectrum of antiviral activity that targeted the nucleocapsidgene of RSV In vivo BALBc murine studies demonstratedthat intranasal dosing of ALN-RSV01 resulted in a 25- to30-log-unit reduction in RSV lung concentration To scaleup this molecule for RSV treatment in humans the safetytolerability and pharmacokinetics were tested on healthyadults demonstrating its safety and tolerance in humansubjects [154] In the human clinical trials of ALN-RSV01healthy subjects were grouped and administered either aplacebo or ALN-RSV01 nasal spray for RSV There was 44reduction in the RSV infection in the subjects who receivedALN-RSV01 without any adverse effect Thus this study inreal terms has established a unique ldquoproof-of-conceptrdquo for anRNAi therapeutic agent in RSV treatment [155] ALN-RSV01proved to be safe and was effective against RSV even in acomplex clinical situation like lung transplants which was aremarkable achievement [156] It is the product of AlnylamPharmaceuticals Inc and has completed phase IIb clinicaltrials

Another approach of RNAi treatment to combat RSVis to decelerate the adverse effects of RSV mediatedTh2 type immune response because the aggravated hostimmune response is more harmful than RSV infection itself

Particularly in neonates RSV bronchiolitis increases IL-4120572 levels which results in increased Th2 response so anantisense oligomer was synthesised for the local silencingof the IL-4120572 gene Intranasal application of the antisenseoligomers into a neonatal murine model reduced the Th2typemediated pulmonary pathological signs of inflammationand lung dysfunction [203] A combinatorial approach of theanti-sense oligomer against RSV and IL-4120572 would controlRSV infection and the adverse effects of RSV mediatedinflammation

Phosphorodiamidate morpholino oligomers (PMOs) arethe oligomers where the nucleobases are covalently attachedwith the morpholine ring replacing the deoxyribose sugarwhile the phosphodiester bond is replaced by the phosphoro-diamidate linkage [157] Morpholino chemical modificationof RNA can be used as antisense with the advantages ofspecificity in vivo stability and targeted delivery PMOs blockthe target complementary RNA and the target RNA failsto interact with the proteins and thus the RNA functionis hindered This is specifically true for mRNA and itstranslation This phenomenon is different from antisense asthe RNase H is not involved [204] Hence PMOs can bedesigned containing the initiation codon against viruses tobe anti-viral Better intake into the cell and in vivo systemscan be facilitated by conjugating cell penetrating peptide(s)(arginine-rich peptide (RXR) 4XB) to it This approach wasattempted against the RSV-L gene to inhibit RSV in cellularand murine models [205]

45 Ethnobotanicals Various natural and synthetic chemicalcompounds have been screened for their application totreat RSV infections Plants are rich sources of alkaloidssteroids flavonoids and other complex compounds that havemedicinal value andmedicinal plants contribute significantlyto the traditional Indian and Chinese medicine AncientChinese literature has descriptions of plant extract againstrespiratory diseases [206] The exact active compound oraction of the traditional formulations is not understood[207] Now modern assays have made it possible to get aninsight into the mechanism and purification of active plantbased compound Extracts of Lonicera deflexicalyx (ChinJinyinhua) were tested against RSV The active compoundfrom the extract was 3 5-dicaffeoylquinic acid (CJ 4-16-4) which was isolated and purified by a series of chro-matographic processes It is suggested that CJ 4-16-4 is afusion inhibitor and the in vitro and in vivo studies suggestthat it is a more effective RSV inhibitor than ribavirin[208] Cytopathic effect (CPE) assay based screening showedthat 27 of 44 herbs had moderate or potent anti-RSVactivity [206] The plant extracts from Cinnamomum cas-sia Cimicifuga foetida Sheng-Ma-Ge-Gen-Tang (SMGGT)(Shoma-kakkon-to) Xiao-Qing-Long-Tang (Sho-seiryu-toso-cheong-ryong-tang) Ge-Gen-Tang and Ginger (Zingiberofficinale) show anti-RSV activity These extracts probablyinhibit RSV infection by blocking the F protein binding to thecell and someof the extracts even stimulate IFN-120573production[158ndash163] Some decoctions like modified Dingchuan (con-sists of Salviae miltiorrhizae radix Scutellariae radix Farfaraeflos and Ephedrae herba) Liu-He-Tang (consist of 13 plant


extract) and water extracts of Licorice (Radix glycyrrhizaeand Radix glycyrrhizae Preparata) have shown effectivenessagainst RSV in vitro Moreover the modified Dingchuandecoction (MDD) exhibited anti-inflammatory and anti-viraleffect in mice (SPF ICR mice) infected with RSV MDDsuppressed eotaxin IL-4 and IFN-120574 level in serum andmRNA expression of TLR4 and NF-120581B in lungs of RSVinfected mice [164]

5 Challenges in the DiagnosisPrevention and Treatment of RSV

Technology has provided enough capabilities for the detec-tion of RSV in various sample types at various stages ofinfection using an array of techniques but the challenge isavailability of these facilities at the correct time for a reason-able cost The most important perspective of RSV diagnosisis the strategic management of choice between the pointof care testing and central laboratory testing [36] Thoughthe point of care testing gives rapid detection advantageit suffers from lower sensitivity and thus is a problem tobe dealt with Rapid diagnostic tools like RT-PCRESI-MSmicroarray based semiautomated respiratory virus nucleicacid test (VRNAT) and the fully automated respiratory virusnucleic acid test SP (RVNATSP) (Nanosphere NorthbrookIL) have proved their efficiency but their application inroutine clinical practice is still a challenge

There are numerous molecules that can be potentialantiviral drugs but the screening of a vast number ofcompounds is cumbersome hence high throughput filteringis an essential part of drug development As an examplewhen 313816 compounds from theMolecular Libraries SmallMolecule Repository were screened against RSV in HEp-2cell line only 409 compounds showed 50 inhibition of thecytopathic effects [209]The challenge after this sophisticatedscreening is the translation into drugs by clearing the phasesof in vivo animal studies and human trials A setback forthe development of therapies against RSV is the lack of agood animal model as they do not truly manifest effects ofRSV infection as in humans RSV experiments in variousanimal models like BALBc mice cotton rats macaquesAfrican green monkeys owl monkeys cebus monkeys bon-net monkeys olive baboons and chimpanzees are evidentin the literature Small animal models like BALBc miceand cotton rats are commonly used due to ease of handlingand low cost whereas the primate studies are conductedwith more stringent regulations and bear heavy expenses[210 211] There are many aspects that need to be addressedin the challenges for vaccine development programs andthe technological interventions to deal with RSV [212]These challenges include safety issues concerning the subjectsinvolved in clinical trials as evident by the failure of formalininactivated RSV vaccine andmotavizumab at the clinical triallevels which resulted in undesired immunogenic responses inthe patients involved [34 116]

Though the present data conclude that RSV is one of theleading causes of morbidity and mortality in children andelders there is no significant correlation between increaseddisease severity respiratory deaths and detection of any

of the respiratory viruses [6 213] The situation is moreintricate as some authors report that coinfection with non-RSV respiratory viruses tends to increase RSV severity [214]and also there is a hypothesis of a synergistic association ofRSV with other viral or bacterial infections [215] Thus thistopic is a subject of debate and these contradictions needclarification and consensus for proper treatment options Alink between atopy asthma and RSV was suspected for avery long time but now there is supporting evidence for thispossible relationship [216ndash218] and is also ascribed partly togenetic factors of RSV and the host [70 71 219 220] Theactivity of RSV in the community may be affected by manyfactors including climate air pollution [221] raceethnicity[222] and social behavior of the population Under thesecomplex factors associated with RSV it is established thatearly detection of risk factors and medical intervention canreduce the incidence of RSV [223] A balance of executionandor abeyance of prophylactic measures is critical withrespect to the above discussed determinants [224] There islimited data to correlate RSV global transmission dynamicswith climate and population [2] due to which it is difficult todevelop strategies for RSV prevention and treatment

6 Conclusions

Currently there is no vaccine or effective treatment againstRSV but the rapid and sensitive RSV detection is possibleThe detection techniques are ameliorated by incorporatingone or more methods and with the advancement in materialscience and biophysical capabilities it has reinforced thedevelopment and design of RSV detection systems Howeveran effective detection technique can be transformed intoeffective diagnosis by integrating it into the communityhealth monitoring program at a reasonable cost Preventionof RSV infection at present is limited to only high riskindividuals with a limited efficacy New preventive measuresresearch like DNA vaccines subunit vaccines and nano-vaccines have reached animal trials On the other handthe RSV treatment approaches using antisense oligomersfusion inhibitors and benzimidazole drug have proceededinto clinical trials The challenges associated with RSVmanagement are categorically numerous However at thecurrent pace of scientific research and development and withthe implementation of scientific commercial and programrecommendations to develop epidemiological strategies itseems optimistic to have an effective diagnosis preventionand treatment solution for RSV in near future

Competing Interests

The authors declare that they have no competing interests

Acknowledgments

The authors acknowledge the National Science FoundationGrant NSF-CREST (HRD-1241701) and HBCU-UP (HRD-1135863) They thank Eva Dennis for the figures used in thispaper


References

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[2] K Bloom-Feshbach W J Alonso V Charu et al ldquoLatitudinalvariations in seasonal activity of influenza and respiratorysyncytial virus (RSV) a global comparative reviewrdquo PLoS ONEvol 8 no 2 Article ID e54445 2013

[3] H Henrickson ldquoCost-effective use of rapid diagnostic tech-niques in the treatment and prevention of viral respiratoryinfectionsrdquo Pediatric Annals vol 34 no 1 pp 24ndash31 2005

[4] L R Krilov ldquoRespiratory syncytial virus disease update ontreatment and preventionrdquo Expert Review of Anti-InfectiveTherapy vol 9 no 1 pp 27ndash32 2011

[5] A Galindo-Fraga A A Ortiz-Hernandez A Ramırez-Venegaset al ldquoClinical characteristics and outcomes of influenza andother influenza-like illnesses in Mexico cityrdquo InternationalJournal of Infectious Diseases vol 17 no 7 pp 510ndash517 2013

[6] D N Tran T M H Pham M T Ha et al ldquoMolecular epidemi-ology and disease severity of human respiratory syncytial virusin Vietnamrdquo PLoS ONE vol 8 no 1 Article ID e45436 2013

[7] P L Collins ldquoRespiratory syncytial virusmdashhuman (Paramyx-oviridae)rdquo in Encyclopedia of Virology G Allan and G WRobert Eds pp 1479ndash1487 Elsevier Oxford UK 2nd edition1999

[8] D Hacking and J Hull ldquoRespiratory syncytial virusmdashviralbiology and the host responserdquo Journal of Infection vol 45 no1 pp 18ndash24 2002

[9] P L Ogra ldquoRespiratory syncytial virus the virus the diseaseand the immune responserdquo Paediatric Respiratory Reviews vol5 pp S119ndashS126 2004

[10] H M Costello W C Ray S Chaiwatpongsakorn and ME Peeples ldquoTargeting RSV with vaccines and small moleculedrugsrdquo Infectious Disorders vol 12 no 2 pp 110ndash128 2012

[11] X Zhao M Singh V N Malashkevich and P S Kim ldquoStruc-tural characterization of the human respiratory syncytial virusfusion protein corerdquo Proceedings of the National Academy ofSciences of theUnited States of America vol 97 no 26 pp 14172ndash14177 2000

[12] P L Collins and B S Graham ldquoViral and host factors in humanrespiratory syncytial virus pathogenesisrdquo Journal of Virologyvol 82 no 5 pp 2040ndash2055 2008

[13] D L Kasper Harrisonrsquos Principles of Internal MedicineMcGraw-Hill 2005

[14] J M Langley J C LeBlanc E E L Wang et al ldquoNosocomialrespiratory syncytial virus infection in Canadian pediatrichospitals a pediatric investigators collaborative network oninfections in Canada studyrdquo Pediatrics vol 100 no 6 pp 943ndash946 1997

[15] A Simon AMuller K Khurana et al ldquoNosocomial infection arisk factor for a complicated course in children with respiratorysyncytial virus infectionmdashresults from a prospective multicen-ter German surveillance studyrdquo International Journal of Hygieneand Environmental Health vol 211 no 3-4 pp 241ndash250 2008

[16] A R Falsey P A Hennessey M A Formica C Cox and EE Walsh ldquoRespiratory syncytial virus infection in elderly andhigh-risk adultsrdquoTheNew England Journal ofMedicine vol 352no 17 pp 1749ndash1759 2005

[17] R Singleton N Etchart S Hou and L Hyland ldquoInability toevoke a long-lasting protective immune response to respiratory

syncytial virus infection inmice correlates with ineffective nasalantibody responsesrdquo Journal of Virology vol 77 no 21 pp11303ndash11311 2003

[18] R D Pockett D Campbell S Carroll F Rajoriya andN Adlard ldquoRotavirus respiratory syncytial virus and non-rotaviral gastroenteritis analysis of hospital readmissions inEngland and Walesrdquo Acta Paediatrica vol 102 no 4 pp e158ndashe163 2013

[19] A R Falsey ldquoRespiratory syncytial virus a global pathogen inan aging worldrdquo Clinical Infectious Diseases vol 57 no 8 pp1078ndash1080 2013

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[21] C Rebuffo-Scheer M Bose J He et al ldquoWhole genomesequencing and evolutionary analysis of human respiratorysyncytial virus A and B from Milwaukee WI 1998-2010rdquo PLoSONE vol 6 no 10 Article ID e25468 2011

[22] C S Khor I C Sam P SHooi andY F Chan ldquoDisplacement ofpredominant respiratory syncytial virus genotypes in Malaysiabetween 1989 and 2011rdquo Infection Genetics and Evolution vol14 pp 357ndash360 2013

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[27] P K Chattopadhyay D A Price T F Harper et al ldquoQuantumdot semiconductor nanocrystals for immunophenotyping bypolychromatic flow cytometryrdquo Nature Medicine vol 12 no 8pp 972ndash977 2006

[28] R A Tripp R Alvarez B Anderson L Jones C Weeksand W Chen ldquoBioconjugated nanoparticle detection of res-piratory syncytial virus infectionrdquo International Journal ofNanomedicine vol 2 no 1 pp 117ndash124 2007

[29] E L Bentzen F House T J Utley J E Crowe Jr and D WWright ldquoProgression of respiratory syncytial virus infectionmonitored by fluorescent quantum dot probesrdquo Nano Lettersvol 5 no 4 pp 591ndash595 2005

[30] D Mattanovich and N Borth ldquoApplications of cell sorting inbiotechnologyrdquoMicrobial Cell Factories vol 5 article 12 2006

[31] M Chen J S Chang M Nason et al ldquoA flow cytometry-based assay to assess RSV-specific neutralizing antibody isreproducible efficient and accuraterdquo Journal of ImmunologicalMethods vol 362 no 1-2 pp 180ndash184 2010

[32] P J Santangelo B Nix A Tsourkas and G Bao ldquoDual FRETmolecular beacons for mRNA detection in living cellsrdquo NucleicAcids Research vol 32 no 6 article e57 2004

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mRNA in live cellsrdquo Journal of the American Chemical Societyvol 132 no 28 pp 9789ndash9796 2010

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[36] P von Lode ldquoPoint-of-care immunotesting approaching theanalytical performance of central laboratory methodsrdquo ClinicalBiochemistry vol 38 no 7 pp 591ndash606 2005

[37] R J Harbeck J Teague G R Crossen D M Maul and PL Childers ldquoNovel rapid optical immunoassay technique fordetection of group A streptococci from pharyngeal specimenscomparison with standard culture methodsrdquo Journal of ClinicalMicrobiology vol 31 no 4 pp 839ndash844 1993

[38] W K Aldous K Gerber E W Taggart J Rupp J Wintchand J A Daly ldquoA comparison of Thermo Electron RSV OIA toviral culture and direct fluorescent assay testing for respiratorysyncytial virusrdquo Journal of Clinical Virology vol 32 no 3 pp224ndash228 2005

[39] R Slinger RMilk I Gaboury and FDiaz-Mitoma ldquoEvaluationof the QuickLab RSV test a new rapid lateral-flow immunoas-say for detection of respiratory syncytial virus antigenrdquo Journalof Clinical Microbiology vol 42 no 8 pp 3731ndash3733 2004

[40] D Gregson T Lloyd S Buchan and D Church ldquoComparisonof theRSVRespi-Stripwith direct fluorescent-antigen detectionfor diagnosis of respiratory syncytial virus infection in pediatricpatientsrdquo Journal of Clinical Microbiology vol 43 no 11 pp5782ndash5783 2005

[41] A P Borek S H Clemens V K Gaskins D Z Aird andA Valsamakis ldquoRespiratory syncytial virus detection by remelXpect BinaxNowRSV direct immunofluorescent staining andtissue culturerdquo Journal of ClinicalMicrobiology vol 44 no 3 pp1105ndash1107 2006

[42] R Selvarangan D Abel and M Hamilton ldquoComparison ofBD Directigen EZ RSV and Binax NOW RSV tests for rapiddetection of respiratory syncytial virus from nasopharyngealaspirates in a pediatric populationrdquoDiagnosticMicrobiology andInfectious Disease vol 62 no 2 pp 157ndash161 2008

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[45] J Mahony S Chong D Bulir A Ruyter K Mwawasi and DWaltho ldquoDevelopment of a sensitive loop-mediated isothermalamplification (LAMP) assay providing specimen-to-result diag-nosis of RSV infections in thirty minutesrdquo Journal of ClinicalMicrobiology vol 51 no 8 pp 2696ndash2701 2013

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transcription-PCR to viral culture and serology in adults withrespiratory illnessrdquo Journal of Clinical Microbiology vol 40 no3 pp 817ndash820 2002

[48] A Hu M Colella J S Tam R Rappaport and S-M ChengldquoSimultaneous detection subgrouping and quantitation ofrespiratory syncytial virus A and B by real-time PCRrdquo Journalof Clinical Microbiology vol 41 no 1 pp 149ndash154 2003

[49] G Dewhurst-Maridor V Simonet J E Bornand L P Nicodand J C Pache ldquoDevelopment of a quantitative TaqMan RT-PCR for respiratory syncytial virusrdquo Journal of VirologicalMethods vol 120 no 1 pp 41ndash49 2004

[50] M Gueudin A Vabret J Petitjean S Gouarin J Brouardand F Freymuth ldquoQuantitation of respiratory syncytial virusRNA in nasal aspirates of children by real-time RT-PCR assayrdquoJournal of Virological Methods vol 109 no 1 pp 39ndash45 2003

[51] R Mentel U Wegner R Bruns and L Gurtler ldquoReal-timePCR to improve the diagnosis of respiratory syncytial virusinfectionrdquo Journal of Medical Microbiology vol 52 no 10 pp893ndash896 2003

[52] L J R van Elden AM van Loon A van der Beek et al ldquoAppli-cability of a real-time quantitative PCR assay for diagnosis ofrespiratory syncytial virus infection in immunocompromisedadultsrdquo Journal of Clinical Microbiology vol 41 no 9 pp 4378ndash4381 2003

[53] J Kuypers N Wright and R Morrow ldquoEvaluation of quanti-tative and type-specific real-time RT-PCR assays for detectionof respiratory syncytial virus in respiratory specimens fromchildrenrdquo Journal of Clinical Virology vol 31 no 2 pp 123ndash1292004

[54] X S Chi F Li J S Tam R Rappaport and S-M Cheng ldquoSemi-quantitative one-step RT-PCR for simultaneous identificationof human influenza and respiratory syncytial virusesrdquo Journalof Virological Methods vol 139 no 1 pp 90ndash92 2007

[55] M L Choudhary S P Anand M Heydari et al ldquoDevelopmentof amultiplex one stepRT-PCR that detects eighteen respiratoryviruses in clinical specimens and comparisonwith real timeRT-PCRrdquo Journal of Virological Methods vol 189 no 1 pp 15ndash192013

[56] K Alby E B Popowitch and M B Miller ldquoComparativeevaluation of the Nanosphere Verigene RV+ assay with theSimplexa Flu AB amp RSV Kit for the detection of influenza andrespiratory syncytial virusesrdquo Journal of Clinical Microbiologyvol 51 no 1 pp 352ndash353 2012

[57] M Xu X Qin M L Astion et al ldquoImplementation of FilmAr-ray respiratory viral panel in a core laboratory improves testingturnaround time and patient carerdquo American Journal of ClinicalPathology vol 139 no 1 pp 118ndash123 2013

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National Academy of Sciences of the United States of Americavol 99 no 24 pp 15687ndash15692 2002

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[66] E C Chen S A Miller J L Derisi and C Y Chiu ldquoUsing apan-viral microarray assay (virochip) to screen clinical samplesfor viral pathogensrdquo Journal of Visualized Experiments vol 27no 50 Article ID e2536 2011

[67] CWWong C LWHeng LWanYee et al ldquoOptimization andclinical validation of a pathogen detectionmicroarrayrdquoGenomeBiology vol 8 no 5 article R93 2007

[68] Y C T Huang Z Li X Hyseni et al ldquoIdentification ofgene biomarkers for respiratory syncytial virus infection in abronchial epithelial cell linerdquo Genomic Medicine vol 2 no 3-4pp 113ndash125 2008

[69] R Janssen J Pennings H Hodemaekers et al ldquoHost tran-scription profiles upon primary respiratory syncytial virusinfectionrdquo Journal of Virology vol 81 no 11 pp 5958ndash59672007

[70] L B Bacharier R Cohen T Schweiger et al ldquoDeterminantsof asthma after severe respiratory syncytial virus bronchiolitisrdquoJournal of Allergy and Clinical Immunology vol 130 no 1 pp91ndash100 2012

[71] N Krishnamoorthy A Khare T B Oriss et al ldquoEarly infec-tion with respiratory syncytial virus impairs regulatory T cellfunction and increases susceptibility to allergic asthmardquoNatureMedicine vol 18 no 10 pp 1525ndash1530 2012

[72] X Yu N Schneiderhan-Marra and T O Joos ldquoProteinmicroarrays for personalizedmedicinerdquoClinical Chemistry vol56 no 3 pp 376ndash387 2010

[73] H Zhu S Hu G Jona et al ldquoSevere acute respiratory syndromediagnostics using a coronavirus protein microarrayrdquo Proceed-ings of the National Academy of Sciences of the United States ofAmerica vol 103 no 11 pp 4011ndash4016 2006

[74] L Yang S Guo Y Li S Zhou and S Tao ldquoProtein microarraysfor systems biologyrdquo Acta Biochimica et Biophysica Sinica vol43 no 3 pp 161ndash171 2011

[75] R Sampath K L Russell C Massire et al ldquoGlobal surveillanceof emerging Influenza virus genotypes by mass spectrometryrdquoPLoS ONE vol 2 no 5 article e489 2007

[76] K F Chen L Blyn R E Rothman et al ldquoReverse transcriptionpolymerase chain reaction and electrospray ionization massspectrometry for identifying acute viral upper respiratory tractinfectionsrdquo Diagnostic Microbiology and Infectious Disease vol69 no 2 pp 179ndash186 2011

[77] M C Demirel S Kao N Malvadkar et al ldquoBio-organism sens-ing via surface enhanced Raman spectroscopy on controlledmetalpolymer nanostructured substratesrdquo Biointerphases vol4 no 2 pp 35ndash41 2009

[78] R A Tripp R A Dluhy and Y Zhao ldquoNovel nanostructures forSERS biosensingrdquo Nano Today vol 3 no 3-4 pp 31ndash37 2008

[79] S D Hudson and G Chumanov ldquoBioanalytical applications ofSERS (surface-enhanced Raman spectroscopy)rdquo Analytical andBioanalytical Chemistry vol 394 no 3 pp 679ndash686 2009

[80] H Liu L Zhang X Lang et al ldquoSingle molecule detectionfrom a large-scale SERS-active Au79Ag21 substraterdquo ScientificReports vol 1 article 112 2011

[81] J M Benevides S A Overman and G J Thomas JrldquoRaman polarized Raman and ultraviolet resonance Ramanspectroscopy of nucleic acids and their complexesrdquo Journal ofRaman Spectroscopy vol 36 no 4 pp 279ndash299 2005

[82] S L Hennigan J D Driskell R A Dluhy et al ldquoDetection ofMycoplasma pneumoniaein simulated and true clinical throatswab specimens by nanorod array-surface-enhanced ramanspectroscopyrdquo PLoS ONE vol 5 no 10 Article ID e13633 2010

[83] J D Driskell Y Zhu C D Kirkwood Y Zhao R A Dluhy andR A Tripp ldquoRapid and sensitive detection of rotavirus molec-ular signatures using surface enhanced raman spectroscopyrdquoPLoS ONE vol 5 no 4 Article ID e10222 2010

[84] S Shanmukh L Jones Y-P Zhao J D Driskell R A Trippand R A Dluhy ldquoIdentification and classification of respiratorysyncytial virus (RSV) strains by surface-enhanced Ramanspectroscopy andmultivariate statistical techniquesrdquoAnalyticaland Bioanalytical Chemistry vol 390 no 6 pp 1551ndash1555 2008

[85] J B McGivney E Bishop K Miller et al ldquoEvaluation of asynthetic peptide as a replacement for the recombinant fusionprotein of respiratory syncytial virus in a potency ELISArdquoJournal of Pharmaceutical and Biomedical Analysis vol 54 no3 pp 572ndash576 2011

[86] G Siuzdak ldquoProbing viruses with mass spectrometryrdquo Journalof Mass Spectrometry vol 33 no 3 pp 203ndash211 1998

[87] B Bothner and G Siuzdak ldquoElectrospray ionization of a wholevirus analyzing mass structure and viabilityrdquo ChemBioChemvol 5 no 3 pp 258ndash260 2004

[88] R Aebersold and M Mann ldquoMass spectrometry-based pro-teomicsrdquo Nature vol 422 no 6928 pp 198ndash207 2003

[89] Y P Ho and P Muralidhar Reddy ldquoIdentification of pathogensby mass spectrometryrdquo Clinical Chemistry vol 56 no 4 pp525ndash536 2010

[90] T C Chou W Hsu C-H Wang Y-J Chen and J-M FangldquoRapid and specific influenza virus detection by functionalizedmagnetic nanoparticles and mass spectrometryrdquo Journal ofNanobiotechnology vol 9 article 52 2011

[91] Z P Yao P A Demirev and C Fenselau ldquoMass spectrometry-based proteolytic mapping for rapid virus identificationrdquo Ana-lytical Chemistry vol 74 no 11 pp 2529ndash2534 2002

[92] A M Caliendo ldquoMultiplex PCR and emerging technologiesfor the detection of respiratory pathogensrdquo Clinical InfectiousDiseases vol 52 no 4 pp S326ndashS330 2011

[93] R Sampath N Mulholland L B Blyn et al ldquoComprehensivebiothreat cluster identification by PCRelectrospray-ionizationmass spectrometryrdquo PLoS ONE vol 7 no 6 Article ID e365282012

[94] A E Casiano-Colon B B Hulbert T K Mayer E E Walshand A R Falsey ldquoLack of sensitivity of rapid antigen tests forthe diagnosis of respiratory syncytial virus infection in adultsrdquoJournal of Clinical Virology vol 28 no 2 pp 169ndash174 2003

[95] A K Shetty E Treynor DWHill KM Gutierrez AWarfordandE J Baron ldquoComparison of conventional viral cultureswithdirect fluorescent antibody stains for diagnosis of community-acquired respiratory virus infections in hospitalized childrenrdquoThe Pediatric Infectious Disease Journal vol 22 no 9 pp 789ndash794 2003


[96] J Aslanzadeh X Zheng H Li et al ldquoProspective evaluation ofrapid antigen tests for diagnosis of respiratory syncytial virusand human metapneumovirus infectionsrdquo Journal of ClinicalMicrobiology vol 46 no 5 pp 1682ndash1685 2008

[97] S A Ali J E Gern T V Hartert et al ldquoReal-world comparisonof two molecular methods for detection of respiratory virusesrdquoVirology Journal vol 8 article 332 2011

[98] P J Jannetto B W Buchan K A Vaughan et al ldquoReal-timedetection of influenza A influenza B and respiratory syncytialvirus A and B in respiratory specimens by use of nanoparticleprobesrdquo Journal of Clinical Microbiology vol 48 no 11 pp3997ndash4002 2010

[99] C S Thaxton D G Georganopoulou and C A Mirkin ldquoGoldnanoparticle probes for the detection of nucleic acid targetsrdquoClinica Chimica Acta vol 363 no 1-2 pp 120ndash126 2006

[100] C B Hall and R G Douglas Jr ldquoModes of transmission ofrespiratory syncytial virusrdquo Journal of Pediatrics vol 99 no 1pp 100ndash103 1981

[101] M G Ottolini S R Curtis A Mathews S R Ottolini andG A Prince ldquoPalivizumab is highly effective in suppressingrespiratory syncytial virus in an immunosuppressed animalmodelrdquoBoneMarrowTransplantation vol 29 no 2 pp 117ndash1202002

[102] K Huang L Incognito X Cheng N D Ulbrandt and H WuldquoRespiratory syncytial virus-neutralizing monoclonal antibod-ies motavizumab and palivizumab inhibit fusionrdquo Journal ofVirology vol 84 no 16 pp 8132ndash8140 2010

[103] C Harkensee M Brodlie N D Embleton and M MckeanldquoPassive immunisation of preterm infants with palivizumabagainst RSV infectionrdquo Journal of Infection vol 52 no 1 pp 2ndash82006

[104] Committee on Infectious Diseases ldquoModified recommenda-tions for use of Palivizumab for prevention of respiratorysyncytial virus infectionsrdquo Pediatrics vol 124 no 6 pp 1694ndash1701 2009

[105] L Zeitlin O Bohorov N Bohorova et al ldquoProphylacticand therapeutic testing of Nicotiana-derived RSV-neutralizinghuman monoclonal antibodies in the cotton rat modelrdquoMAbsvol 5 no 2 pp 263ndash269 2013

[106] X Li S Sambhara C X Li et al ldquoPlasmid DNA encodingthe respiratory syncytial virus G protein is a promising vaccinecandidaterdquo Virology vol 269 no 1 pp 54ndash65 2000

[107] X Li S Sambhara C X Li et al ldquoProtection against respiratorysyncytial virus infection by DNA immunizationrdquoThe Journal ofExperimental Medicine vol 188 no 4 pp 681ndash688 1998

[108] N Ternette B Tippler K Uberla and T Grunwald ldquoImmuno-genicity and efficacy of codon optimized DNA vaccines encod-ing the F-protein of respiratory syncytial virusrdquoVaccine vol 25no 41 pp 7271ndash7279 2007

[109] L Martinez-Sobrido N Gitiban A Fernandez-Sesma et alldquoProtection against respiratory syncytial virus by a recombinantNewcastle disease virus vectorrdquo Journal of Virology vol 80 no3 pp 1130ndash1139 2006

[110] HWuVADennis S R Pillai and S R Singh ldquoRSV fusion (F)protein DNA vaccine provides partial protection against viralinfectionrdquo Virus Research vol 145 no 1 pp 39ndash47 2009

[111] A M Talaat R Lyons and S A Johnston ldquoA combination vac-cine confers full protection against co-infections with influenzaherpes simplex and respiratory syncytial virusesrdquo Vaccine vol20 no 3-4 pp 538ndash544 2001

[112] S M Bueno P A Gonzalez K M Cautivo et al ldquoProtectiveT cell immunity against respiratory syncytial virus is efficientlyinduced by recombinant BCGrdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 105 no52 pp 20822ndash20827 2008

[113] C Xie J S He M Zhang et al ldquoOral respiratory syncytialvirus (RSV) DNA vaccine expressing RSV F protein deliveredby attenuated Salmonella typhimuriumrdquo Human Gene Therapyvol 18 no 8 pp 746ndash752 2007

[114] Y H Fu J S He X B Wang et al ldquoA prime-boost vacci-nation strategy using attenuated Salmonella typhimurium anda replication-deficient recombinant adenovirus vector elicitsprotective immunity against human respiratory syncytial virusrdquoBiochemical and Biophysical Research Communications vol 395no 1 pp 87ndash92 2010

[115] M B Elliott T Chen N B Terio et al ldquoAlphavirus repliconparticles encoding the fusion or attachment glycoproteins ofrespiratory syncytial virus elicit protective immune responsesin BALBc mice and functional serum antibodies in rhesusmacaquesrdquo Vaccine vol 25 no 41 pp 7132ndash7144 2007

[116] H W Kim J G Canchola C D Brandt et al ldquoRespiratorysyncytial virus disease in infants despite prior administrationof antigenic inactivated vaccinerdquo American Journal of Epidemi-ology vol 89 no 4 pp 422ndash434 1969

[117] P J M Openshaw and J S Tregoning ldquoImmune responsesand disease enhancement during respiratory syncytial virusinfectionrdquo Clinical Microbiology Reviews vol 18 no 3 pp 541ndash555 2005

[118] J F Valarcher and G Taylor ldquoBovine respiratory syncytial virusinfectionrdquo Veterinary Research vol 38 no 2 pp 153ndash180 2007

[119] S van Drunen Littel-van den Hurk J W Mapletoft N Arsicand J Kovacs-Nolan ldquoImmunopathology of RSV infectionprospects for developing vaccines without this complicationrdquoReviews in Medical Virology vol 17 no 1 pp 5ndash34 2007

[120] S R Singh V A Dennis C L Carter et al ldquoImmunogenicityand efficacy of recombinant RSV-F vaccine in a mouse modelrdquoVaccine vol 25 no 33 pp 6211ndash6223 2007

[121] P Subbarayan H Qin S Pillai et al ldquoExpression and charac-terization of a multivalent human respiratory syncytial virusproteinrdquo Molekuliarnaia Biologiia vol 44 no 3 pp 477ndash4872010

[122] M R Murawski L W McGinnes R W Finberg et alldquoNewcastle disease virus-like particles containing respiratorysyncytial virus G protein induced protection in BALBc micewith no evidence of immunopathologyrdquo Journal of Virology vol84 no 2 pp 1110ndash1123 2010

[123] T Takimoto J L Hurwitz C Coleclough et al ldquoRecombinantSendai virus expressing the G glycoprotein of respiratorysyncytial virus (RSV) elicits immune protection against RSVrdquoJournal of Virology vol 78 no 11 pp 6043ndash6047 2004

[124] B G Jones R E Sealy R Rudraraju et al ldquoSendai virus-based RSV vaccine protects African green monkeys from RSVinfectionrdquo Vaccine vol 30 no 5 pp 959ndash968 2012

[125] T M Ward V Traina-Dorge K A Davis and W L GrayldquoRecombinant simian varicella viruses expressing respiratorysyncytial virus antigens are immunogenicrdquo Journal of GeneralVirology vol 89 no 3 pp 741ndash750 2008

[126] M Chen K-F Hu B Rozell C Orvell B Morein andP Liljestrom ldquoVaccination with recombinant alphavirus orimmune-stimulating complex antigen against respiratory syn-cytial virusrdquo Journal of Immunology vol 169 no 6 pp 3208ndash3216 2002


[127] S Boyoglu K Vig S Pillai et al ldquoEnhanced delivery andexpression of a nanoencapsulated DNA vaccine vector forrespiratory syncytial virusrdquoNanomedicine vol 5 no 4 pp 463ndash472 2009

[128] S B Barnum P Subbarayan K Vig et al ldquoNano-EncapsulatedDNAandor protein boost immunizations increase efficiency ofDNA vaccine protection against RSVrdquo Journal of Nanomedicineand Nanotechnology vol 3 article 312 2012

[129] E Eroglu P M Tiwari A B Waffo et al ldquoA nonviralpHEMA+chitosan nanosphere-mediated high-efficiency genedelivery systemrdquo International Journal of Nanomedicine vol 8pp 1403ndash1415 2013

[130] X Roux C Dubuquoy G Durand et al ldquoSub-nucloecapsidnanoparticles a nasal vaccine against respiratory syncytialvirusrdquo PLoS ONE vol 3 no 3 Article ID e1766 2008

[131] D M Lindell S B Morris M P White et al ldquoA novel inac-tivated intranasal respiratory syncytial virus vaccine promotesviral clearance without TH2 associated Vaccine-Enhanced dis-easerdquo PLoS ONE vol 6 no 7 Article ID e21823 2011

[132] G Smith R Raghunandan Y Wu et al ldquoRespiratory syncytialvirus fusion glycoprotein expressed in insect cells form proteinnanoparticles that induce protective immunity in cotton ratsrdquoPLoS ONE vol 7 no 11 Article ID e50852 2012

[133] GMGlenn G Smith L Fries et al ldquoSafety and immunogenic-ity of a Sf9 insect cell-derived respiratory syncytial virus fusionprotein nanoparticlerdquo Vaccine vol 31 no 3 pp 524ndash532 2013

[134] F M Ferolla D R Hijano P L Acosta et al ldquoMacronutrientsduring pregnancy and life-threatening respiratory syncytialvirus infections in childrenrdquo American Journal of Respiratoryand Critical Care Medicine vol 187 no 9 pp 983ndash990 2013

[135] N Zang X Xie Y Deng et al ldquoResveratrol-mediated gammainterferon reduction prevents airway inflammation and air-way hyperresponsiveness in respiratory syncytial virus-infectedimmunocompromisedmicerdquo Journal of Virology vol 85 no 24pp 13061ndash13068 2011

[136] X H Xie N Zang S M Li et al ldquoReseveratrol inhibits re-spiratory syncytial virus-induced IL-6 production decreasesvirla replication and downregulates TRIF expression in airwayepithelial cellsrdquo Inflammation vol 35 no 4 pp 1392ndash1401 2012

[137] M E Belderbos M L Houben B Wilbrink et al ldquoCord bloodvitamin D deficiency is associated with respiratory syncytialvirus bronchiolitisrdquo Pediatrics vol 127 no 6 pp e1513ndashe15202011

[138] S Hansdottir M M Monick N Lovan L Powers A Gerkeand G W Hunninghake ldquoVitamin D decreases respiratorysyncytial virus induction of NF-120581B-linked chemokines andcytokines in airway epithelium while maintaining the antiviralstaterdquo Journal of Immunology vol 184 no 2 pp 965ndash974 2010

[139] N Al-Sonboli N Al-Aghbari A Al-Aryani et al ldquoMicronu-trient concentrations in respiratory syncytial virus and humanmetapneumovirus in yemeni childrenrdquo Annals of TropicalPaediatrics vol 29 no 1 pp 35ndash40 2009

[140] S F Dowell Z Papic J S Bresee et al ldquoTreatment ofrespiratory syncytial virus infection with vitamin A a random-ized placebo-controlled trial in Santiago Chilerdquo The PediatricInfectious Disease Journal vol 15 no 9 pp 782ndash786 1996

[141] E Chiba Y Tomosada M G Vizoso-Pinto et al ldquoImmuno-biotic Lactobacillus rhamnosus improves resistance of infantmice against respiratory syncytial virus infectionrdquo InternationalImmunopharmacology vol 17 no 2 pp 373ndash382 2013

[142] J Z Wu C C Lin and Z Hong ldquoRibavirin viramidineand adenosine-deaminase-catalysed drug activation implica-tion for nucleoside prodrug designrdquo Journal of AntimicrobialChemotherapy vol 52 no 4 pp 543ndash546 2003

[143] R G Gish ldquoTreating HCV with ribavirin analogues andribavirin-like moleculesrdquo Journal of Antimicrobial Chemother-apy vol 57 no 1 pp 8ndash13 2006

[144] B Langhans H D Nischalke S Arndt et al ldquoRibavirinexerts differential effects on functions of CD4+Th1 Th2 andregulatory T cell clones in hepatitis Crdquo PLoS ONE vol 7 no7 Article ID e42094 2012

[145] W Markland T J Mcquaid J Jain and A D Kwong ldquoBroad-spectrum antiviral activity of the IMP dehydrogenase inhibitorVX- 497 a comparison with ribavirin and demonstration ofantiviral additivity with alpha interferonrdquo Antimicrobial Agentsand Chemotherapy vol 44 no 4 pp 859ndash866 2000

[146] J Chapman E Abbott D G Alber et al ldquoRSV604 a novelinhibitor of respiratory syncytial virus replicationrdquo Antimicro-bial Agents and Chemotherapy vol 51 no 9 pp 3346ndash33532007

[147] L Ni L Zhao Y Qian et al ldquoDesign and characterization ofhuman respiratory syncytial virus entry inhibitorsrdquo AntiviralTherapy vol 10 no 7 pp 833ndash840 2005

[148] M K Pastey T L Gower P W Spearman J E Crowe Jr andB S Graham ldquoA RhoA-derived peptide inhibits syncytium for-mation induced by respiratory syncytial virus andparainfluenzavirus type 3rdquo Nature Medicine vol 6 no 1 pp 35ndash40 2000

[149] X A Wang C W Cianci K L Yu et al ldquoRespiratory syncytialvirus fusion inhibitors Part 5 optimization of benzimidazolesubstitution patterns towards derivatives with improved activ-ityrdquo Bioorganic and Medicinal Chemistry Letters vol 17 no 16pp 4592ndash4598 2007

[150] A A Nikitenko Y E Raifeld and T Z Wang ldquoThe discoveryof RFI-641 as a potent and selective inhibitor of the respiratorysyncytial virusrdquo Bioorganic and Medicinal Chemistry Lettersvol 11 no 8 pp 1041ndash1044 2001

[151] L Sun A K Singh K Vig S R Pillai and S R Singh ldquoSilvernanoparticles inhibit replication of respiratory syncytial virusrdquoJournal of Biomedical Nanotechnology vol 4 no 2 pp 149ndash1582008

[152] S R Singh P M Tiwari and V A Dennis ldquoAnti-respiratorysyncytial virus peptide functionalized gold nanoparticlesrdquo USPatent Office 2012

[153] R Alvarez S Elbashir T Borland et al ldquoRNA interference-mediated silencing of the respiratory syncytial virus nucleocap-sid defines a potent antiviral strategyrdquoAntimicrobial Agents andChemotherapy vol 53 no 9 pp 3952ndash3962 2009

[154] J DeVincenzo J E Cehelsky R Alvarez et al ldquoEvaluation ofthe safety tolerability and pharmacokinetics of ALN-RSV01 anovel RNAi antiviral therapeutic directed against respiratorysyncytial virus (RSV)rdquoAntiviral Research vol 77 no 3 pp 225ndash231 2008

[155] J DeVincenzo R Lambkin-Williams T Wilkinson et alldquoA randomized double-blind placebo-controlled study of anRNAi-based therapy directed against respiratory syncytialvirusrdquo Proceedings of the National Academy of Sciences of theUnited States of America vol 107 no 19 pp 8800ndash8805 2010

[156] M R Zamora M Budev M Rolfe et al ldquoRNA interferencetherapy in lung transplant patients infected with respiratorysyncytial virusrdquo American Journal of Respiratory and CriticalCare Medicine vol 183 no 4 pp 531ndash538 2011


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[158] K C Wang J S Chang L C Chiang and C-C LinldquoSheng-Ma-Ge-Gen-Tang (Shoma-kakkon-to) inhibited cyto-pathic effect of human respiratory syncytial virus in cell linesof human respiratory tractrdquo Journal of Ethnopharmacology vol135 no 2 pp 538ndash544 2011

[159] K CWang J S Chang L C Chiang andC C Lin ldquoCimicifugafoetida L inhibited human respiratory syncytial virus in HEp-2and A549 cell linesrdquoThe American Journal of Chinese Medicinevol 40 no 1 pp 151ndash162 2012

[160] J S Chang C F Yeh K C Wang D E Shieh M H Yen andL C Chiang ldquoXiao-Qing-Long-Tang (Sho-seiryu-to) inhibitedcytopathic effect of human respiratory syncytial virus in celllines of human respiratory tractrdquo Journal of Ethnopharmacologyvol 147 no 2 pp 481ndash487 2013

[161] J S Chang K CWang C F Yeh D E Shieh and L C ChiangldquoFresh ginger (Zingiber officinale) has anti-viral activity againsthuman respiratory syncytial virus in human respiratory tractcell linesrdquo Journal of Ethnopharmacology vol 145 no 1 pp 146ndash151 2013

[162] J S Chang K CWang D E Shieh F F Hsu and L C ChiangldquoGe-Gen-Tang has anti-viral activity against human respiratorysyncytial virus in human respiratory tract cell linesrdquo Journal ofEthnopharmacology vol 139 no 1 pp 305ndash310 2012

[163] C F Yeh J S Chang K C Wang D E Shieh and L CChiang ldquoWater extract ofCinnamomum cassia Blume inhibitedhuman respiratory syncytial virus by preventing viral attach-ment internalization and syncytium formationrdquo Journal ofEthnopharmacology vol 147 no 2 pp 321ndash326 2013

[164] L Li C H Yu H Z Ying and J M Yu ldquoAntiviral effectsof modified Dingchuan decoction against respiratory syncytialvirus infection in vitro and in an immunosuppressive mousemodelrdquo Journal of Ethnopharmacology vol 147 no 1 pp 238ndash244 2013

[165] J H Connor M O McKenzie G D Parks and D S LylesldquoAntiviral activity and RNA polymerase degradation followingHsp90 inhibition in a range of negative strand virusesrdquoVirologyvol 362 no 1 pp 109ndash119 2007

[166] R Geller R Andino and J Frydman ldquoHsp90 inhibitors exhibitresistance-free antiviral activity against respiratory syncytialvirusrdquo PLoS ONE vol 8 no 2 Article ID e56762 2013

[167] C Wild T Greenwell and T Matthews ldquoA synthetic peptidefrom HIV-1 gp41 is a potent inhibitor of virus-mediated cell-cell fusionrdquo AIDS Research and Human Retroviruses vol 9 no11 pp 1051ndash1053 1993

[168] E Wang X Sun Y Qian L Zhao P Tien and G F Gao ldquoBothheptad repeats of human respiratory syncytial virus fusionprotein are potent inhibitors of viral fusionrdquo Biochemical andBiophysical Research Communications vol 302 no 3 pp 469ndash475 2003

[169] K N Bossart B A Mungall G Crameri L F Wang B TEaton and C C Broder ldquoInhibition of Henipavirus fusion andinfection by heptad-derived peptides of the Nipah virus fusionglycoproteinrdquo Virology Journal vol 2 article 57 2005

[170] M Porotto L Doctor P Carta et al ldquoInhibition ofHendra virusfusionrdquo Journal of Virology vol 80 no 19 pp 9837ndash9849 2006

[171] M Porotto P Carta Y Deng et al ldquoMolecular determinants ofantiviral potency of paramyxovirus entry inhibitorsrdquo Journal ofVirology vol 81 no 19 pp 10567ndash10574 2007

[172] M Porotto B Rockx C C Yokoyama et al ldquoInhibitionof Nipah virus infection in vivo targeting an early stage ofparamyxovirus fusion activation during viral entryrdquo PLoS Path-ogens vol 6 no 10 Article ID e1001168 2010

[173] C Deffrasnes M-E Hamelin G A Prince and G BoivinldquoIdentification and evaluation of a highly effective fusioninhibitor for human metapneumovirusrdquo Antimicrobial Agentsand Chemotherapy vol 52 no 1 pp 279ndash287 2008

[174] D M Lambert S Barney A L Lambert et al ldquoPeptides fromconserved regions of paramyxovirus fusion (F) proteins arepotent inhibitors of viral fusionrdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 93 no5 pp 2186ndash2191 1996

[175] K Andries M Moeremans T Gevers et al ldquoSubstitutedbenzimidazoles with nanomolar activity against respiratorysyncytial virusrdquo Antiviral Research vol 60 no 3 pp 209ndash2192003

[176] A Lundin T Bergstrom L Bendrioua N Kann B Adamiakand E Trybala ldquoTwo novel fusion inhibitors of human respira-tory syncytial virusrdquo Antiviral Research vol 88 no 3 pp 317ndash324 2010

[177] K L Yu Y Zhang R L Civiello et al ldquoFundamental structure-activity relationships associated with a new structural class ofrespiratory syncytial virus inhibitorrdquo Bioorganic and MedicinalChemistry Letters vol 13 no 13 pp 2141ndash2144 2003

[178] K L Yu Y Zhang R L Civiello et al ldquoRespiratory syncytialvirus inhibitors Part 2 benzimidazol-2-one derivativesrdquo Bioor-ganic and Medicinal Chemistry Letters vol 14 no 5 pp 1133ndash1137 2004

[179] K L Yu X A Wang R L Civiello et al ldquoRespiratory syncytialvirus fusion inhibitors Part 3 water-soluble benzimidazol-2-one derivatives with antiviral activity in vivordquo Bioorganic andMedicinal Chemistry Letters vol 16 no 5 pp 1115ndash1122 2006

[180] KD CombrinkH B Gulgeze JWThuring et al ldquoRespiratorysyncytial virus fusion inhibitors Part 6 an examination of theeffect of structural variation of the benzimidazol-2-one hetero-cycle moietyrdquo Bioorganic and Medicinal Chemistry Letters vol17 no 17 pp 4784ndash4790 2007

[181] N Sin B L Venables K D Combrink et al ldquoRespiratorysyncytial virus fusion inhibitors Part 7 structure-activityrelationships associated with a series of isatin oximes thatdemonstrate antiviral activity in vivordquoBioorganic andMedicinalChemistry Letters vol 19 no 16 pp 4857ndash4862 2009

[182] A Nikitenko Y Raifeld B Mitsner and H Newman ldquoPyrimi-dine containing RSV fusion inhibitorsrdquo Bioorganic and Medici-nal Chemistry Letters vol 15 no 2 pp 427ndash430 2005

[183] D C Pryde T D Tran I Gardner et al ldquoNon-benzimidazolecontaining inhibitors of respiratory syncytial virusrdquo BioorganicandMedicinal Chemistry Letters vol 23 no 3 pp 827ndash833 2013

[184] S Galdiero A Falanga M Vitiello M Cantisani V Marra andM Galdiero ldquoSilver nanoparticles as potential antiviral agentsrdquoMolecules vol 16 no 10 pp 8894ndash8918 2011

[185] P M Tiwari K Vig V A Dennis and S R Singh ldquoFunction-alized gold nanoparticles and their biomedical applicationsrdquoNanomaterials vol 1 no 1 pp 31ndash63 2011

[186] S Vardharajula S Z Ali PM Tiwari et al ldquoFunctionalized car-bon nanotubes biomedical applicationsrdquo International Journalof Nanomedicine vol 7 pp 5361ndash5374 2012

[187] J W Stone N J Thornburg D L Blum S J Kuhn DW Wright and J E Crowe Jr ldquoGold nanorod vaccine forrespiratory syncytial virusrdquo Nanotechnology vol 24 no 29Article ID 295102 2013


[188] N S Lee T Dohjima G Bauer et al ldquoExpression of smallinterfering RNAs targeted against HIV-1 rev transcripts inhuman cellsrdquo Nature Biotechnology vol 20 no 5 pp 500ndash5052002

[189] S Barik ldquoControl of nonsegmented negative-strand RNA virusreplication by siRNArdquo Virus Research vol 102 no 1 pp 27ndash352004

[190] Y L Zhang T Cheng Y J Cai et al ldquoRNA interference inhibitshepatitis B virus of different genotypes in vitro and in vivordquoBMCMicrobiology vol 10 article 214 2010

[191] S Jairath P Brown Vargas H A Hamlin A K Field and R EKilkuskie ldquoInhibition of respiratory syncytial virus replicationby antisense oligodeoxyribonucleotidesrdquo Antiviral Researchvol 33 no 3 pp 201ndash213 1997

[192] V Bitko AMusiyenko O Shulyayeva and S Barik ldquoInhibitionof respiratory viruses by nasally administered siRNArdquo NatureMedicine vol 11 no 1 pp 50ndash55 2005

[193] V Bitko and S Barik ldquoPhenotypic silencing of cytoplasmicgenes using sequence-specific double-stranded short interfer-ing RNA and its application in the reverse genetics of wild typenegative-strand RNA virusesrdquo BMCMicrobiology vol 1 article34 2001

[194] K Vig N Lewis E G Moore S Pillai V A Dennis andS R Singh ldquoSecondary RNA structure and its role in RNAinterference to silence the respiratory syncytial virus fusionprotein generdquo Molecular Biotechnology vol 43 no 3 pp 200ndash211 2009

[195] M Kumar S S Mohapatra A K Behera et al ldquoIntranasalgene transfer by chitosan-DNA nanospheres protects BALBcmice against acute respiratory syncytial virus infectionrdquoHumanGene Therapy vol 13 no 12 pp 1415ndash1425 2002

[196] W Zhang H Yang X Kong et al ldquoInhibition of respiratorysyncytial virus infection with intranasal siRNA nanoparticlestargeting the viral NS1 generdquo Nature Medicine vol 11 no 1 pp56ndash62 2005

[197] B Dong and R H Silverman ldquo2-5A-dependent RNasemolecules dimerize during activation by 2-5Ardquo The Journal ofBiological Chemistry vol 270 no 8 pp 4133ndash4137 1995

[198] NM Cirino G LiW Xiao P F Torrence and R H SilvermanldquoTargeting RNA decay with 2101584051015840 oligoadenylate-antisense inrespiratory syncytial virus-infected cellsrdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 94 no 5 pp 1937ndash1942 1997

[199] D L Barnard R W Sidwell W Xiao M R Player S AAdah and P F Torrence ldquo2-5A-DNA conjugate inhibition ofrespiratory syncytial virus replication effects of oligonucleotidestructuremodifications and RNA target site selectionrdquoAntiviralResearch vol 41 no 3 pp 119ndash134 1999

[200] M R Player D L Barnard and P F Torrence ldquoPotentinhibition of respiratory syncytial virus replication using a2-5A-antisense chimera targeted to signals within the virusgenomic RNArdquo Proceedings of the National Academy of Sciencesof the United States of America vol 95 no 15 pp 8874ndash88791998

[201] Z Xu M Kuang J R Okicki H Cramer and N ChaudharyldquoPotent inhibition of respiratory syncytial virus by combina-tion treatment with 2-5A antisense and ribavirinrdquo AntiviralResearch vol 61 no 3 pp 195ndash206 2004

[202] DW Leaman F J Longano J ROkicki et al ldquoTargeted therapyof respiratory syncytial virus in African greenmonkeys by intranasally administered 2-5A antisenserdquo Virology vol 292 no 1pp 70ndash77 2002

[203] M J Ripple D You S Honnegowda et al ldquoImmunomodu-lation with IL-4R120572 antisense oligonucleotide prevents respira-tory syncytial virus-mediated pulmonary diseaserdquo Journal ofImmunology vol 185 no 8 pp 4804ndash4811 2010

[204] J Summerton ldquoMorpholino antisense oligomers the case for anRNaseH-independent structural typerdquoBiochimica et BiophysicaActa vol 1489 no 1 pp 141ndash158 1999

[205] S H Lai D A Stein A Guerrero-Plata et al ldquoInhibition of res-piratory syncytial virus infections with morpholino oligomersin cell cultures and in micerdquo Molecular Therapy vol 16 no 6pp 1120ndash1128 2008

[206] S C Ma J Du P P-H But et al ldquoAntiviral Chinese medicinalherbs against respiratory syncytial virusrdquo Journal of Ethnophar-macology vol 79 no 2 pp 205ndash211 2002

[207] R Vaidya ldquoObservational therapeutics scope challenges andorganizationrdquo Journal of Ayurveda and IntegrativeMedicine vol2 no 4 pp 165ndash169 2011

[208] J O Ojwang Y H Wang P R Wyde et al ldquoA novel inhibitorof respiratory syncytial virus isolated from ethnobotanicalsrdquoAntiviral Research vol 68 no 3 pp 163ndash172 2005

[209] D H Chung B Moore D Matharu et al ldquoA cell basedhigh-throughput screening approach for the discovery of newinhibitors of respiratory syncytial virusrdquo Virology Journal vol10 article 19 2013

[210] U F Power ldquoRespiratory syncytial virus (RSV) vaccines-twosteps back for one leap forwardrdquo Journal of Clinical Virology vol41 no 1 pp 38ndash44 2008

[211] J F Papin R F Wolf S D Kosanke et al ldquoInfant Baboonsinfected with respiratory syncytial virus develop clinical andpathologic changes that parallel those of human infantsrdquoAmerican Journal of PhysiologymdashLung Cellular and MolecularPhysiology vol 304 no 8 pp L530ndashL539 2013

[212] B S Graham ldquoBiological challenges and technological oppor-tunities for respiratory syncytial virus vaccine developmentrdquoImmunological Reviews vol 239 no 1 pp 149ndash166 2011

[213] L J Stockman W A Brooks P K Streatfield et al ldquoChal-lenges to evaluating respiratory syncytial virus mortality inBangladesh 2004-2008rdquo PLoS ONE vol 8 no 1 Article IDe53857 2013

[214] Y Harada F Kinoshita L M Yoshida et al ldquoDoes respira-tory virus co-infection increase the clinical severity of acuterespiratory infection among children infected with respiratorysyncytial virusrdquoThePediatric Infectious Disease Journal vol 32no 5 pp 441ndash445 2013

[215] A A T M Bosch G Biesbroek K Trzcinski E A M Sandersand D Bogaert ldquoViral and bacterial interactions in the upperrespiratory tractrdquo PLoS Pathogens vol 9 no 1 Article IDe1003057 2013

[216] P G Holt and P D Sly ldquoInteractions between RSV infectionasthma and atopy unraveling the complexitiesrdquoThe Journal ofExperimental Medicine vol 196 no 10 pp 1271ndash1275 2002

[217] J Han K Takeda and EW Gelfand ldquoThe role of RSV infectionin asthma initiation and progression findings in a mousemodelrdquo Pulmonary Medicine vol 2011 Article ID 748038 8pages 2011

[218] S M Szabo A R Levy K L Gooch et al ldquoElevated riskof asthma after hospitalization for respiratory syncytial virusinfection in infancyrdquo Paediatric Respiratory Reviews vol 13 no2 pp 70161ndash70166 2013

[219] L I Tapia S AmpueroM A Palomino et al ldquoRespiratory syn-cytial virus infection and recurrentwheezing inChilean infants


a genetic backgroundrdquo Infection Genetics and Evolution vol16 no 0 pp 54ndash61 2013

[220] S FThomsen S van der Sluis L G Stensballe et al ldquoExploringthe association between severe respiratory syncytial virus infec-tion and asthma a registry-based twin studyrdquoAmerican Journalof Respiratory and Critical Care Medicine vol 179 no 12 pp1091ndash1097 2009

[221] S Vandini L Corvaglia R Alessandroni et al ldquoRespiratorysyncytial virus infection in infants and correlation with meteo-rological factors and air pollutantsrdquo Italian Journal of Pediatricsvol 39 article 1 2013

[222] M K Iwane S S Chaves P G Szilagyi et al ldquoDisparitiesbetween black and white children in hospitalizations associ-ated with acute respiratory illness and laboratory-confirmedinfluenza and respiratory syncytial virus in 3 US Counties-2002-2009rdquo American Journal of Epidemiology vol 177 no 7pp 656ndash665 2013

[223] A A El Kholy N AMostafa S A El-Sherbini et al ldquoMorbidityand outcome of severe respiratory syncytial virus infectionrdquoPediatrics International vol 14 no 10 pp 283ndash288 2013

[224] R C Welliver Sr ldquoTemperature humidity and ultraviolet Bradiation predict community respiratory syncytial virus activ-ityrdquoThe Pediatric Infectious Disease Journal vol 26 supplement11 pp S29ndashS35 2007

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of


Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 2014

Zoology


Molecular Biology International

GenomicsInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014


BioinformaticsAdvances in

Marine BiologyJournal of



Signal TransductionJournal of


BioMed Research International

Evolutionary BiologyInternational Journal of



Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Genetics Research International


Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational



Enzyme Research



Microbiology


G protein

N protein

F protein

(a) (b)

P protein L protein

SH protein

Single stranded RNA

Matrix protein

NS1

NS2

N

P

M

SH

G

F

M2

L

Figure 1 Structure and genome organization of respiratory syncytial virus (a) Approximately 200 nm RSV virion particle and (b) singlestranded negative RNA genome consisting of 10 genes

necessary for the fusion budding and spread of the virus[9 10] After attachment to the host cell RSV fuses withthe host cell membrane using the F protein through the6 helix coiled-coil bundle of the F protein a mechanismcharacteristically found in paramyxoviridae members [11]Although the detailed mechanism of RSV infection is notfully understood the most accepted mechanism is the entryof the nucleocapsid into the host cell mediated by the Fprotein through clathrin mediated endocytosis [12] TheRNA is first converted into a plus strand which serves asthe template for replication whereas for transcription theRNA genome itself transcribes mRNA for protein synthesiswithout any intermediate

Almost all children of 2 years of age will have had an RSVinfection and leading to 160000ndash600000 deaths per year [4]Approximately 25 to 40 of infants and children at thefirst exposure to RSV have signs or symptoms of bronchiolitisor pneumonia These symptoms include rhinorrhea low-grade fever cough and wheezing The symptoms in adultsmay include common cold with rhinorrhea sore throatcough malaise headache and fever It can also lead toexacerbated symptoms such as severe pneumonia in theelderly especially residing in nursing homes [13] Usuallychildren show symptoms within 4 to 6 days of infectionand most of them recover in 1 to 2 weeks while serving

as carriers of the virus for 1 to 3 weeks RSV infectionin children of nosocomial origin is associated with highermortality than community-acquired illness because of thepre-existingmorbidity [14 15] SevereRSVdisease risk hoversfor the elderly and adults with chronic heart or lung diseaseor with weakened immune system [16] RSV infection doesnot provoke lasting immunity [17] therefore reinfection isvery common [18] Recently RSV infection was reported toaccount for hospitalizations and mortality in elderly people[19] RSV accounted for severe lower respiratory tract infec-tions including chronic lung disease systemic comorbiditiesand even death At present there is no specific treatmentfor RSV infection ever since its first discovery in 1956 [20]Currently Food and Drug Administration (FDA) approvedprophylactic drug for RSV that includes palivizumab andribavirin administered along with symptomatic treatmentdrugs and supportive care Currently techniques used fordiagnosis of RSV include ELISA direct immunofluorescencewestern blot PCR and real-time PCR The diagnosis andtreatment scenario has significantly changed with the adventof advanced techniques and in-depth understanding of RSVbiology but the execution of these clinical developmentsin practice requires extensive study and time This reviewpresents the recent advances in the diagnosis prevention andtreatment of RSV (Figure 2)



Quantum dots

Immuno-PCR

Microarray


SERS

LFIA

ELISA

Immunoassays

OIA

DFA

LAMP

Real-time PCR

DNA based assays

PCR-ESI-MS

Microarray

DNA vaccines

Subunit vaccines

Prevention

AntibodiesNutrition

Nanovaccines


Treatment

Antisense RNA


Diagnosis


2 Diagnosis






ble1Com

paris

onof


es

Techniqu

eRe

ference

Principle

Advantages

Drawbacks

Currentu

sage

status

(A)F

luorescenceb

ased

metho

ds

(1)D

FA[2425]

Microscop

icdetectionof

RSVwith

specifica

ntibod

yconjugated

with

fluorop

hore

Easy

procedure

Hum

anerrorfading

ofdyes

Research

intentH

ospitalbased

procedurecommercial

diagno

sticassays

(2)Q

Ds

[26ndash

31]

Detectio

nof

signalsfro

mflu

orescent

nano

particles

upon

encoun

terw

ithRS

Veither

throug

hmicroscop

yor

flow

cytometry



Toxicityinsolub

ility

Research

intent

(3)M

olecular

beacon

based

imaging

[3233]

Hairpin

DNAfunctio

nalized

gold

nano

particlewith

fluorop

hore

hybridizationwith

targetmRN

ALive

cellim

agingwith

real-timed

etectio

nProb

ablegene

silencing


Research

intent

(B)Immun

oassays

(1)E

LISA

[3435]

Specificb

inding

andcolorim

etric

detectionof

antig

en-antibod

ycomplex

Easy


sensitivity

Cumbersom

epron

etohu

man

errors

Hospitalbased

procedure

commercialdiagno

sticassays

(2)O

IA[36ndash

38]

Presence

ofspecifica

ntigen-antibod

ycomplex

form

edaltersther

eflectiv

esurfa

cesp

ropertiesw

hich

isvisually

detected


Needs

confi

rmationby

other

tests

forn

egatives

amples

Research

intentnot

commercialized

(3)L

FIA

[39ndash

42]

Immun

o-complexes

detected

chromatograph

ically


Aapproved

Non

quantitativelim

itof

sample

volumelim

itsdetection

Hospitalbased

procedure

commercialdiagno

sticassays

(C)M

olecular

metho

ds

(1)L

AMP

[4143ndash4

5]Colorim

etric

turbidim

etric

detectionof

isothermalam

plificatio

nof

DNAusing

specificp

rimer


Semiquantitativ

edesig

ning

compatib

leprim

erset

Research

intentnot

commercialized

(2)P

CR[4647]

Amplificatio

nof

viralcDNAand

visualizationof

PCRprod

uct

Rapidandsensitive

than

conventio

nal

cultu

remetho

dsHighlim

itsof

detection

Research

intenthospitalbased

procedure

(3)R

eal-T

ime

PCR

[48ndash53]

Real-timea

mplificatio

nof

targetDNAor

cDNA

Rapid(3ndash5


and

very

lowlim

itsof

detection

Expensive

Research

intenthospitalbased


(4)M

ultip

lex

PCR

[54ndash

57]

Use

ofmultip

leprim

erandor

prob

esets

Simultaneou

sdetectio

nof

multip


rstrains

Lesssensitive

Research

intenthospitalbased

procedure

(5)Immun

o-PC

R[5859]

Acombinatio

nof

immun

oassay

and

real-timeP

CR

Very

lowlim

itsof

detection

improved

limits

ofdetectionover

individu

alEL

ISA

andPC

R(400


)Com

plex

experim

entald

esign

Research

intentnot

commercialized

(6)M

icroarray

[60ndash

74]

Hybrid

izationof

sampleb

iomolecules

toim

mob

ilizedtargetDNAor

proteinon

achip


scaleidentificatio

nof

multip


nucle

icacid

targets

Cost-ineffective

Research

intenthospitalbased


(D)B

ioph

ysicalmetho

d

(1)P

CR-ESI-M

S[7576]

Massspectroscop

yof

PCR-am

plicon

sthroug

hele

ctronspraydispersio

n

Highlysensitive

andspecifice

venat

strain

leveland

efficientm

ultip

lepathogensd

etectio

nEx

pensive

Research

intentnot

commercialized

(2)S

ERS

[77ndash84]

Inelastic

scatterin

gof

mon

ochrom

atic

radiationup

oninteractionwith

ananalytew

ithlow-fr

equencyvibrational

andor

rotatio

nalenergy

Rapidandno

ndestructiv

edetectio

nof

analytes

with

high

sensitivity

Samplep

reparatio

nRe

search

intentnot

commercialized






























PCRamplicon



Silver vaporsource


Spectra


Laser

Silver nanorods













3 Prevention




Antigenome


Protein synthesis

AssemblySH protein

N protein

Nucleus

P protein

L protein


Golgi complex




F protein

G protein



+

+

+

+

+

Gold nanoparticles

Antibodies




RSV

RSV

RSV

RSV

Drug



No infection

No infection

No infection

Binding

Cell












Naked DNAvector

Vaccination

RSV


RSV

RSV challenge

RSV challenge


Mice protected

Expression

DNA vaccine

Viral DNA

Vector

vector

















4 Treatment








Table2Com

paris

onof

different

treatmentapp

roachesfor

RSV

Treatm

ent

Mechanism

Exam

ple

Remark

References

Antivira

ldrugs


nRS

V60

4Eff

ectiv

eagainstRS

Vbu

tadverse

effecto

ntheh

ost

[104]

[142ndash146]

Mutation

Ribavirin

vira

midinemerim

epod

ibInhibitoro

fino

sinem

onop

hosphate

dehydrogenase

Ribavirin

mycop

heno

latem

izoribine

Immun

ostim

ulatoryeffects

Ribavirin

Fusio

ninhibitors

Inhibitin

gfusio


Peptidemdash

HR121H

R212R

hoA

Chem

icalmdashBM

S-433771R

FI-641


ising

anti


ninhibitorshave

sidee

ffects

[147ndash150]

Nanop

articles

Inhibitin

gattachmenttocell

Silver

nano

particlesgold

nano

particles

Emerging

fieldcon

clusiv

estudies

requ

ired

[151152]

Antise

nsetherapy

RNAinterfe

rence

siRNA-ALN

-RSV

01Ph

osph

orod

iamidatem

orph

olinooligom

ersEff

ectiv

eand

safeA

LN-RSV

01completed

phaseIIb

clinicaltrails

[153ndash157]

Ethn

obotanicals

Prob

ablyfusio

ninhibitors

anti-inflammatory

Plantextractsmdash

Cinn

amom

umcassia

Cimicifuga

foetida


Decoctio

ns-m

odified

Dingchu

an

Liu-He-Tang

water

extracto

fLicorice

Prom

ising

butcon

clusiv

estudies

requ

ired

[158ndash164

]























6 Conclusions


Competing Interests


Acknowledgments



References




















































































































































































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology



Quantum dots

Immuno-PCR

Microarray


SERS

LFIA

ELISA

Immunoassays

OIA

DFA

LAMP

Real-time PCR

DNA based assays

PCR-ESI-MS

Microarray

DNA vaccines

Subunit vaccines

Prevention

AntibodiesNutrition

Nanovaccines


Treatment

Antisense RNA


Diagnosis


2 Diagnosis






ble1Com

paris

onof


es

Techniqu

eRe

ference

Principle

Advantages

Drawbacks

Currentu

sage

status

(A)F

luorescenceb

ased

metho

ds

(1)D

FA[2425]

Microscop

icdetectionof

RSVwith

specifica

ntibod

yconjugated

with

fluorop

hore

Easy

procedure

Hum

anerrorfading

ofdyes

Research

intentH

ospitalbased

procedurecommercial

diagno

sticassays

(2)Q

Ds

[26ndash

31]

Detectio

nof

signalsfro

mflu

orescent

nano

particles

upon

encoun

terw

ithRS

Veither

throug

hmicroscop

yor

flow

cytometry



Toxicityinsolub

ility

Research

intent

(3)M

olecular

beacon

based

imaging

[3233]

Hairpin

DNAfunctio

nalized

gold

nano

particlewith

fluorop

hore

hybridizationwith

targetmRN

ALive

cellim

agingwith

real-timed

etectio

nProb

ablegene

silencing


Research

intent

(B)Immun

oassays

(1)E

LISA

[3435]

Specificb

inding

andcolorim

etric

detectionof

antig

en-antibod

ycomplex

Easy


sensitivity

Cumbersom

epron

etohu

man

errors

Hospitalbased

procedure

commercialdiagno

sticassays

(2)O

IA[36ndash

38]

Presence

ofspecifica

ntigen-antibod

ycomplex

form

edaltersther

eflectiv

esurfa

cesp

ropertiesw

hich

isvisually

detected


Needs

confi

rmationby

other

tests

forn

egatives

amples

Research

intentnot

commercialized

(3)L

FIA

[39ndash

42]

Immun

o-complexes

detected

chromatograph

ically


Aapproved

Non

quantitativelim

itof

sample

volumelim

itsdetection

Hospitalbased

procedure

commercialdiagno

sticassays

(C)M

olecular

metho

ds

(1)L

AMP

[4143ndash4

5]Colorim

etric

turbidim

etric

detectionof

isothermalam

plificatio

nof

DNAusing

specificp

rimer


Semiquantitativ

edesig

ning

compatib

leprim

erset

Research

intentnot

commercialized

(2)P

CR[4647]

Amplificatio

nof

viralcDNAand

visualizationof

PCRprod

uct

Rapidandsensitive

than

conventio

nal

cultu

remetho

dsHighlim

itsof

detection

Research

intenthospitalbased

procedure

(3)R

eal-T

ime

PCR

[48ndash53]

Real-timea

mplificatio

nof

targetDNAor

cDNA

Rapid(3ndash5


and

very

lowlim

itsof

detection

Expensive

Research

intenthospitalbased


(4)M

ultip

lex

PCR

[54ndash

57]

Use

ofmultip

leprim

erandor

prob

esets

Simultaneou

sdetectio

nof

multip


rstrains

Lesssensitive

Research

intenthospitalbased

procedure

(5)Immun

o-PC

R[5859]

Acombinatio

nof

immun

oassay

and

real-timeP

CR

Very

lowlim

itsof

detection

improved

limits

ofdetectionover

individu

alEL

ISA

andPC

R(400


)Com

plex

experim

entald

esign

Research

intentnot

commercialized

(6)M

icroarray

[60ndash

74]

Hybrid

izationof

sampleb

iomolecules

toim

mob

ilizedtargetDNAor

proteinon

achip


scaleidentificatio

nof

multip


nucle

icacid

targets

Cost-ineffective

Research

intenthospitalbased


(D)B

ioph

ysicalmetho

d

(1)P

CR-ESI-M

S[7576]

Massspectroscop

yof

PCR-am

plicon

sthroug

hele

ctronspraydispersio

n

Highlysensitive

andspecifice

venat

strain

leveland

efficientm

ultip

lepathogensd

etectio

nEx

pensive

Research

intentnot

commercialized

(2)S

ERS

[77ndash84]

Inelastic

scatterin

gof

mon

ochrom

atic

radiationup

oninteractionwith

ananalytew

ithlow-fr

equencyvibrational

andor

rotatio

nalenergy

Rapidandno

ndestructiv

edetectio

nof

analytes

with

high

sensitivity

Samplep

reparatio

nRe

search

intentnot

commercialized






























PCRamplicon



Silver vaporsource


Spectra


Laser

Silver nanorods













3 Prevention




Antigenome


Protein synthesis

AssemblySH protein

N protein

Nucleus

P protein

L protein


Golgi complex




F protein

G protein



+

+

+

+

+

Gold nanoparticles

Antibodies




RSV

RSV

RSV

RSV

Drug



No infection

No infection

No infection

Binding

Cell












Naked DNAvector

Vaccination

RSV


RSV

RSV challenge

RSV challenge


Mice protected

Expression

DNA vaccine

Viral DNA

Vector

vector

















4 Treatment








Table2Com

paris

onof

different

treatmentapp

roachesfor

RSV

Treatm

ent

Mechanism

Exam

ple

Remark

References

Antivira

ldrugs


nRS

V60

4Eff

ectiv

eagainstRS

Vbu

tadverse

effecto

ntheh

ost

[104]

[142ndash146]

Mutation

Ribavirin

vira

midinemerim

epod

ibInhibitoro

fino

sinem

onop

hosphate

dehydrogenase

Ribavirin

mycop

heno

latem

izoribine

Immun

ostim

ulatoryeffects

Ribavirin

Fusio

ninhibitors

Inhibitin

gfusio


Peptidemdash

HR121H

R212R

hoA

Chem

icalmdashBM

S-433771R

FI-641


ising

anti


ninhibitorshave

sidee

ffects

[147ndash150]

Nanop

articles

Inhibitin

gattachmenttocell

Silver

nano

particlesgold

nano

particles

Emerging

fieldcon

clusiv

estudies

requ

ired

[151152]

Antise

nsetherapy

RNAinterfe

rence

siRNA-ALN

-RSV

01Ph

osph

orod

iamidatem

orph

olinooligom

ersEff

ectiv

eand

safeA

LN-RSV

01completed

phaseIIb

clinicaltrails

[153ndash157]

Ethn

obotanicals

Prob

ablyfusio

ninhibitors

anti-inflammatory

Plantextractsmdash

Cinn

amom

umcassia

Cimicifuga

foetida


Decoctio

ns-m

odified

Dingchu

an

Liu-He-Tang

water

extracto

fLicorice

Prom

ising

butcon

clusiv

estudies

requ

ired

[158ndash164

]























6 Conclusions


Competing Interests


Acknowledgments



References




















































































































































































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


ble1Com

paris

onof


es

Techniqu

eRe

ference

Principle

Advantages

Drawbacks

Currentu

sage

status

(A)F

luorescenceb

ased

metho

ds

(1)D

FA[2425]

Microscop

icdetectionof

RSVwith

specifica

ntibod

yconjugated

with

fluorop

hore

Easy

procedure

Hum

anerrorfading

ofdyes

Research

intentH

ospitalbased

procedurecommercial

diagno

sticassays

(2)Q

Ds

[26ndash

31]

Detectio

nof

signalsfro

mflu

orescent

nano

particles

upon

encoun

terw

ithRS

Veither

throug

hmicroscop

yor

flow

cytometry



Toxicityinsolub

ility

Research

intent

(3)M

olecular

beacon

based

imaging

[3233]

Hairpin

DNAfunctio

nalized

gold

nano

particlewith

fluorop

hore

hybridizationwith

targetmRN

ALive

cellim

agingwith

real-timed

etectio

nProb

ablegene

silencing


Research

intent

(B)Immun

oassays

(1)E

LISA

[3435]

Specificb

inding

andcolorim

etric

detectionof

antig

en-antibod

ycomplex

Easy


sensitivity

Cumbersom

epron

etohu

man

errors

Hospitalbased

procedure

commercialdiagno

sticassays

(2)O

IA[36ndash

38]

Presence

ofspecifica

ntigen-antibod

ycomplex

form

edaltersther

eflectiv

esurfa

cesp

ropertiesw

hich

isvisually

detected


Needs

confi

rmationby

other

tests

forn

egatives

amples

Research

intentnot

commercialized

(3)L

FIA

[39ndash

42]

Immun

o-complexes

detected

chromatograph

ically


Aapproved

Non

quantitativelim

itof

sample

volumelim

itsdetection

Hospitalbased

procedure

commercialdiagno

sticassays

(C)M

olecular

metho

ds

(1)L

AMP

[4143ndash4

5]Colorim

etric

turbidim

etric

detectionof

isothermalam

plificatio

nof

DNAusing

specificp

rimer


Semiquantitativ

edesig

ning

compatib

leprim

erset

Research

intentnot

commercialized

(2)P

CR[4647]

Amplificatio

nof

viralcDNAand

visualizationof

PCRprod

uct

Rapidandsensitive

than

conventio

nal

cultu

remetho

dsHighlim

itsof

detection

Research

intenthospitalbased

procedure

(3)R

eal-T

ime

PCR

[48ndash53]

Real-timea

mplificatio

nof

targetDNAor

cDNA

Rapid(3ndash5


and

very

lowlim

itsof

detection

Expensive

Research

intenthospitalbased


(4)M

ultip

lex

PCR

[54ndash

57]

Use

ofmultip

leprim

erandor

prob

esets

Simultaneou

sdetectio

nof

multip


rstrains

Lesssensitive

Research

intenthospitalbased

procedure

(5)Immun

o-PC

R[5859]

Acombinatio

nof

immun

oassay

and

real-timeP

CR

Very

lowlim

itsof

detection

improved

limits

ofdetectionover

individu

alEL

ISA

andPC

R(400


)Com

plex

experim

entald

esign

Research

intentnot

commercialized

(6)M

icroarray

[60ndash

74]

Hybrid

izationof

sampleb

iomolecules

toim

mob

ilizedtargetDNAor

proteinon

achip


scaleidentificatio

nof

multip


nucle

icacid

targets

Cost-ineffective

Research

intenthospitalbased


(D)B

ioph

ysicalmetho

d

(1)P

CR-ESI-M

S[7576]

Massspectroscop

yof

PCR-am

plicon

sthroug

hele

ctronspraydispersio

n

Highlysensitive

andspecifice

venat

strain

leveland

efficientm

ultip

lepathogensd

etectio

nEx

pensive

Research

intentnot

commercialized

(2)S

ERS

[77ndash84]

Inelastic

scatterin

gof

mon

ochrom

atic

radiationup

oninteractionwith

ananalytew

ithlow-fr

equencyvibrational

andor

rotatio

nalenergy

Rapidandno

ndestructiv

edetectio

nof

analytes

with

high

sensitivity

Samplep

reparatio

nRe

search

intentnot

commercialized






























PCRamplicon



Silver vaporsource


Spectra


Laser

Silver nanorods













3 Prevention




Antigenome


Protein synthesis

AssemblySH protein

N protein

Nucleus

P protein

L protein


Golgi complex




F protein

G protein



+

+

+

+

+

Gold nanoparticles

Antibodies




RSV

RSV

RSV

RSV

Drug



No infection

No infection

No infection

Binding

Cell












Naked DNAvector

Vaccination

RSV


RSV

RSV challenge

RSV challenge


Mice protected

Expression

DNA vaccine

Viral DNA

Vector

vector

















4 Treatment








Table2Com

paris

onof

different

treatmentapp

roachesfor

RSV

Treatm

ent

Mechanism

Exam

ple

Remark

References

Antivira

ldrugs


nRS

V60

4Eff

ectiv

eagainstRS

Vbu

tadverse

effecto

ntheh

ost

[104]

[142ndash146]

Mutation

Ribavirin

vira

midinemerim

epod

ibInhibitoro

fino

sinem

onop

hosphate

dehydrogenase

Ribavirin

mycop

heno

latem

izoribine

Immun

ostim

ulatoryeffects

Ribavirin

Fusio

ninhibitors

Inhibitin

gfusio


Peptidemdash

HR121H

R212R

hoA

Chem

icalmdashBM

S-433771R

FI-641


ising

anti


ninhibitorshave

sidee

ffects

[147ndash150]

Nanop

articles

Inhibitin

gattachmenttocell

Silver

nano

particlesgold

nano

particles

Emerging

fieldcon

clusiv

estudies

requ

ired

[151152]

Antise

nsetherapy

RNAinterfe

rence

siRNA-ALN

-RSV

01Ph

osph

orod

iamidatem

orph

olinooligom

ersEff

ectiv

eand

safeA

LN-RSV

01completed

phaseIIb

clinicaltrails

[153ndash157]

Ethn

obotanicals

Prob

ablyfusio

ninhibitors

anti-inflammatory

Plantextractsmdash

Cinn

amom

umcassia

Cimicifuga

foetida


Decoctio

ns-m

odified

Dingchu

an

Liu-He-Tang

water

extracto

fLicorice

Prom

ising

butcon

clusiv

estudies

requ

ired

[158ndash164

]























6 Conclusions


Competing Interests


Acknowledgments



References




















































































































































































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology






























PCRamplicon



Silver vaporsource


Spectra


Laser

Silver nanorods













3 Prevention




Antigenome


Protein synthesis

AssemblySH protein

N protein

Nucleus

P protein

L protein


Golgi complex




F protein

G protein



+

+

+

+

+

Gold nanoparticles

Antibodies




RSV

RSV

RSV

RSV

Drug



No infection

No infection

No infection

Binding

Cell












Naked DNAvector

Vaccination

RSV


RSV

RSV challenge

RSV challenge


Mice protected

Expression

DNA vaccine

Viral DNA

Vector

vector

















4 Treatment








Table2Com

paris

onof

different

treatmentapp

roachesfor

RSV

Treatm

ent

Mechanism

Exam

ple

Remark

References

Antivira

ldrugs


nRS

V60

4Eff

ectiv

eagainstRS

Vbu

tadverse

effecto

ntheh

ost

[104]

[142ndash146]

Mutation

Ribavirin

vira

midinemerim

epod

ibInhibitoro

fino

sinem

onop

hosphate

dehydrogenase

Ribavirin

mycop

heno

latem

izoribine

Immun

ostim

ulatoryeffects

Ribavirin

Fusio

ninhibitors

Inhibitin

gfusio


Peptidemdash

HR121H

R212R

hoA

Chem

icalmdashBM

S-433771R

FI-641


ising

anti


ninhibitorshave

sidee

ffects

[147ndash150]

Nanop

articles

Inhibitin

gattachmenttocell

Silver

nano

particlesgold

nano

particles

Emerging

fieldcon

clusiv

estudies

requ

ired

[151152]

Antise

nsetherapy

RNAinterfe

rence

siRNA-ALN

-RSV

01Ph

osph

orod

iamidatem

orph

olinooligom

ersEff

ectiv

eand

safeA

LN-RSV

01completed

phaseIIb

clinicaltrails

[153ndash157]

Ethn

obotanicals

Prob

ablyfusio

ninhibitors

anti-inflammatory

Plantextractsmdash

Cinn

amom

umcassia

Cimicifuga

foetida


Decoctio

ns-m

odified

Dingchu

an

Liu-He-Tang

water

extracto

fLicorice

Prom

ising

butcon

clusiv

estudies

requ

ired

[158ndash164

]























6 Conclusions


Competing Interests


Acknowledgments



References




















































































































































































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology




















PCRamplicon



Silver vaporsource


Spectra


Laser

Silver nanorods













3 Prevention




Antigenome


Protein synthesis

AssemblySH protein

N protein

Nucleus

P protein

L protein


Golgi complex




F protein

G protein



+

+

+

+

+

Gold nanoparticles

Antibodies




RSV

RSV

RSV

RSV

Drug



No infection

No infection

No infection

Binding

Cell












Naked DNAvector

Vaccination

RSV


RSV

RSV challenge

RSV challenge


Mice protected

Expression

DNA vaccine

Viral DNA

Vector

vector

















4 Treatment








Table2Com

paris

onof

different

treatmentapp

roachesfor

RSV

Treatm

ent

Mechanism

Exam

ple

Remark

References

Antivira

ldrugs


nRS

V60

4Eff

ectiv

eagainstRS

Vbu

tadverse

effecto

ntheh

ost

[104]

[142ndash146]

Mutation

Ribavirin

vira

midinemerim

epod

ibInhibitoro

fino

sinem

onop

hosphate

dehydrogenase

Ribavirin

mycop

heno

latem

izoribine

Immun

ostim

ulatoryeffects

Ribavirin

Fusio

ninhibitors

Inhibitin

gfusio


Peptidemdash

HR121H

R212R

hoA

Chem

icalmdashBM

S-433771R

FI-641


ising

anti


ninhibitorshave

sidee

ffects

[147ndash150]

Nanop

articles

Inhibitin

gattachmenttocell

Silver

nano

particlesgold

nano

particles

Emerging

fieldcon

clusiv

estudies

requ

ired

[151152]

Antise

nsetherapy

RNAinterfe

rence

siRNA-ALN

-RSV

01Ph

osph

orod

iamidatem

orph

olinooligom

ersEff

ectiv

eand

safeA

LN-RSV

01completed

phaseIIb

clinicaltrails

[153ndash157]

Ethn

obotanicals

Prob

ablyfusio

ninhibitors

anti-inflammatory

Plantextractsmdash

Cinn

amom

umcassia

Cimicifuga

foetida


Decoctio

ns-m

odified

Dingchu

an

Liu-He-Tang

water

extracto

fLicorice

Prom

ising

butcon

clusiv

estudies

requ

ired

[158ndash164

]























6 Conclusions


Competing Interests


Acknowledgments



References




















































































































































































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology












PCRamplicon



Silver vaporsource


Spectra


Laser

Silver nanorods













3 Prevention




Antigenome


Protein synthesis

AssemblySH protein

N protein

Nucleus

P protein

L protein


Golgi complex




F protein

G protein



+

+

+

+

+

Gold nanoparticles

Antibodies




RSV

RSV

RSV

RSV

Drug



No infection

No infection

No infection

Binding

Cell












Naked DNAvector

Vaccination

RSV


RSV

RSV challenge

RSV challenge


Mice protected

Expression

DNA vaccine

Viral DNA

Vector

vector

















4 Treatment








Table2Com

paris

onof

different

treatmentapp

roachesfor

RSV

Treatm

ent

Mechanism

Exam

ple

Remark

References

Antivira

ldrugs


nRS

V60

4Eff

ectiv

eagainstRS

Vbu

tadverse

effecto

ntheh

ost

[104]

[142ndash146]

Mutation

Ribavirin

vira

midinemerim

epod

ibInhibitoro

fino

sinem

onop

hosphate

dehydrogenase

Ribavirin

mycop

heno

latem

izoribine

Immun

ostim

ulatoryeffects

Ribavirin

Fusio

ninhibitors

Inhibitin

gfusio


Peptidemdash

HR121H

R212R

hoA

Chem

icalmdashBM

S-433771R

FI-641


ising

anti


ninhibitorshave

sidee

ffects

[147ndash150]

Nanop

articles

Inhibitin

gattachmenttocell

Silver

nano

particlesgold

nano

particles

Emerging

fieldcon

clusiv

estudies

requ

ired

[151152]

Antise

nsetherapy

RNAinterfe

rence

siRNA-ALN

-RSV

01Ph

osph

orod

iamidatem

orph

olinooligom

ersEff

ectiv

eand

safeA

LN-RSV

01completed

phaseIIb

clinicaltrails

[153ndash157]

Ethn

obotanicals

Prob

ablyfusio

ninhibitors

anti-inflammatory

Plantextractsmdash

Cinn

amom

umcassia

Cimicifuga

foetida


Decoctio

ns-m

odified

Dingchu

an

Liu-He-Tang

water

extracto

fLicorice

Prom

ising

butcon

clusiv

estudies

requ

ired

[158ndash164

]























6 Conclusions


Competing Interests


Acknowledgments



References




















































































































































































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology




PCRamplicon



Silver vaporsource


Spectra


Laser

Silver nanorods













3 Prevention




Antigenome


Protein synthesis

AssemblySH protein

N protein

Nucleus

P protein

L protein


Golgi complex




F protein

G protein



+

+

+

+

+

Gold nanoparticles

Antibodies




RSV

RSV

RSV

RSV

Drug



No infection

No infection

No infection

Binding

Cell












Naked DNAvector

Vaccination

RSV


RSV

RSV challenge

RSV challenge


Mice protected

Expression

DNA vaccine

Viral DNA

Vector

vector

















4 Treatment








Table2Com

paris

onof

different

treatmentapp

roachesfor

RSV

Treatm

ent

Mechanism

Exam

ple

Remark

References

Antivira

ldrugs


nRS

V60

4Eff

ectiv

eagainstRS

Vbu

tadverse

effecto

ntheh

ost

[104]

[142ndash146]

Mutation

Ribavirin

vira

midinemerim

epod

ibInhibitoro

fino

sinem

onop

hosphate

dehydrogenase

Ribavirin

mycop

heno

latem

izoribine

Immun

ostim

ulatoryeffects

Ribavirin

Fusio

ninhibitors

Inhibitin

gfusio


Peptidemdash

HR121H

R212R

hoA

Chem

icalmdashBM

S-433771R

FI-641


ising

anti


ninhibitorshave

sidee

ffects

[147ndash150]

Nanop

articles

Inhibitin

gattachmenttocell

Silver

nano

particlesgold

nano

particles

Emerging

fieldcon

clusiv

estudies

requ

ired

[151152]

Antise

nsetherapy

RNAinterfe

rence

siRNA-ALN

-RSV

01Ph

osph

orod

iamidatem

orph

olinooligom

ersEff

ectiv

eand

safeA

LN-RSV

01completed

phaseIIb

clinicaltrails

[153ndash157]

Ethn

obotanicals

Prob

ablyfusio

ninhibitors

anti-inflammatory

Plantextractsmdash

Cinn

amom

umcassia

Cimicifuga

foetida


Decoctio

ns-m

odified

Dingchu

an

Liu-He-Tang

water

extracto

fLicorice

Prom

ising

butcon

clusiv

estudies

requ

ired

[158ndash164

]























6 Conclusions


Competing Interests


Acknowledgments



References




















































































































































































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology







3 Prevention




Antigenome


Protein synthesis

AssemblySH protein

N protein

Nucleus

P protein

L protein


Golgi complex




F protein

G protein



+

+

+

+

+

Gold nanoparticles

Antibodies




RSV

RSV

RSV

RSV

Drug



No infection

No infection

No infection

Binding

Cell












Naked DNAvector

Vaccination

RSV


RSV

RSV challenge

RSV challenge


Mice protected

Expression

DNA vaccine

Viral DNA

Vector

vector

















4 Treatment








Table2Com

paris

onof

different

treatmentapp

roachesfor

RSV

Treatm

ent

Mechanism

Exam

ple

Remark

References

Antivira

ldrugs


nRS

V60

4Eff

ectiv

eagainstRS

Vbu

tadverse

effecto

ntheh

ost

[104]

[142ndash146]

Mutation

Ribavirin

vira

midinemerim

epod

ibInhibitoro

fino

sinem

onop

hosphate

dehydrogenase

Ribavirin

mycop

heno

latem

izoribine

Immun

ostim

ulatoryeffects

Ribavirin

Fusio

ninhibitors

Inhibitin

gfusio


Peptidemdash

HR121H

R212R

hoA

Chem

icalmdashBM

S-433771R

FI-641


ising

anti


ninhibitorshave

sidee

ffects

[147ndash150]

Nanop

articles

Inhibitin

gattachmenttocell

Silver

nano

particlesgold

nano

particles

Emerging

fieldcon

clusiv

estudies

requ

ired

[151152]

Antise

nsetherapy

RNAinterfe

rence

siRNA-ALN

-RSV

01Ph

osph

orod

iamidatem

orph

olinooligom

ersEff

ectiv

eand

safeA

LN-RSV

01completed

phaseIIb

clinicaltrails

[153ndash157]

Ethn

obotanicals

Prob

ablyfusio

ninhibitors

anti-inflammatory

Plantextractsmdash

Cinn

amom

umcassia

Cimicifuga

foetida


Decoctio

ns-m

odified

Dingchu

an

Liu-He-Tang

water

extracto

fLicorice

Prom

ising

butcon

clusiv

estudies

requ

ired

[158ndash164

]























6 Conclusions


Competing Interests


Acknowledgments



References




















































































































































































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


Antigenome


Protein synthesis

AssemblySH protein

N protein

Nucleus

P protein

L protein


Golgi complex




F protein

G protein



+

+

+

+

+

Gold nanoparticles

Antibodies




RSV

RSV

RSV

RSV

Drug



No infection

No infection

No infection

Binding

Cell












Naked DNAvector

Vaccination

RSV


RSV

RSV challenge

RSV challenge


Mice protected

Expression

DNA vaccine

Viral DNA

Vector

vector

















4 Treatment








Table2Com

paris

onof

different

treatmentapp

roachesfor

RSV

Treatm

ent

Mechanism

Exam

ple

Remark

References

Antivira

ldrugs


nRS

V60

4Eff

ectiv

eagainstRS

Vbu

tadverse

effecto

ntheh

ost

[104]

[142ndash146]

Mutation

Ribavirin

vira

midinemerim

epod

ibInhibitoro

fino

sinem

onop

hosphate

dehydrogenase

Ribavirin

mycop

heno

latem

izoribine

Immun

ostim

ulatoryeffects

Ribavirin

Fusio

ninhibitors

Inhibitin

gfusio


Peptidemdash

HR121H

R212R

hoA

Chem

icalmdashBM

S-433771R

FI-641


ising

anti


ninhibitorshave

sidee

ffects

[147ndash150]

Nanop

articles

Inhibitin

gattachmenttocell

Silver

nano

particlesgold

nano

particles

Emerging

fieldcon

clusiv

estudies

requ

ired

[151152]

Antise

nsetherapy

RNAinterfe

rence

siRNA-ALN

-RSV

01Ph

osph

orod

iamidatem

orph

olinooligom

ersEff

ectiv

eand

safeA

LN-RSV

01completed

phaseIIb

clinicaltrails

[153ndash157]

Ethn

obotanicals

Prob

ablyfusio

ninhibitors

anti-inflammatory

Plantextractsmdash

Cinn

amom

umcassia

Cimicifuga

foetida


Decoctio

ns-m

odified

Dingchu

an

Liu-He-Tang

water

extracto

fLicorice

Prom

ising

butcon

clusiv

estudies

requ

ired

[158ndash164

]























6 Conclusions


Competing Interests


Acknowledgments



References




















































































































































































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology









Naked DNAvector

Vaccination

RSV


RSV

RSV challenge

RSV challenge


Mice protected

Expression

DNA vaccine

Viral DNA

Vector

vector

















4 Treatment








Table2Com

paris

onof

different

treatmentapp

roachesfor

RSV

Treatm

ent

Mechanism

Exam

ple

Remark

References

Antivira

ldrugs


nRS

V60

4Eff

ectiv

eagainstRS

Vbu

tadverse

effecto

ntheh

ost

[104]

[142ndash146]

Mutation

Ribavirin

vira

midinemerim

epod

ibInhibitoro

fino

sinem

onop

hosphate

dehydrogenase

Ribavirin

mycop

heno

latem

izoribine

Immun

ostim

ulatoryeffects

Ribavirin

Fusio

ninhibitors

Inhibitin

gfusio


Peptidemdash

HR121H

R212R

hoA

Chem

icalmdashBM

S-433771R

FI-641


ising

anti


ninhibitorshave

sidee

ffects

[147ndash150]

Nanop

articles

Inhibitin

gattachmenttocell

Silver

nano

particlesgold

nano

particles

Emerging

fieldcon

clusiv

estudies

requ

ired

[151152]

Antise

nsetherapy

RNAinterfe

rence

siRNA-ALN

-RSV

01Ph

osph

orod

iamidatem

orph

olinooligom

ersEff

ectiv

eand

safeA

LN-RSV

01completed

phaseIIb

clinicaltrails

[153ndash157]

Ethn

obotanicals

Prob

ablyfusio

ninhibitors

anti-inflammatory

Plantextractsmdash

Cinn

amom

umcassia

Cimicifuga

foetida


Decoctio

ns-m

odified

Dingchu

an

Liu-He-Tang

water

extracto

fLicorice

Prom

ising

butcon

clusiv

estudies

requ

ired

[158ndash164

]























6 Conclusions


Competing Interests


Acknowledgments



References




















































































































































































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


Naked DNAvector

Vaccination

RSV


RSV

RSV challenge

RSV challenge


Mice protected

Expression

DNA vaccine

Viral DNA

Vector

vector

















4 Treatment








Table2Com

paris

onof

different

treatmentapp

roachesfor

RSV

Treatm

ent

Mechanism

Exam

ple

Remark

References

Antivira

ldrugs


nRS

V60

4Eff

ectiv

eagainstRS

Vbu

tadverse

effecto

ntheh

ost

[104]

[142ndash146]

Mutation

Ribavirin

vira

midinemerim

epod

ibInhibitoro

fino

sinem

onop

hosphate

dehydrogenase

Ribavirin

mycop

heno

latem

izoribine

Immun

ostim

ulatoryeffects

Ribavirin

Fusio

ninhibitors

Inhibitin

gfusio


Peptidemdash

HR121H

R212R

hoA

Chem

icalmdashBM

S-433771R

FI-641


ising

anti


ninhibitorshave

sidee

ffects

[147ndash150]

Nanop

articles

Inhibitin

gattachmenttocell

Silver

nano

particlesgold

nano

particles

Emerging

fieldcon

clusiv

estudies

requ

ired

[151152]

Antise

nsetherapy

RNAinterfe

rence

siRNA-ALN

-RSV

01Ph

osph

orod

iamidatem

orph

olinooligom

ersEff

ectiv

eand

safeA

LN-RSV

01completed

phaseIIb

clinicaltrails

[153ndash157]

Ethn

obotanicals

Prob

ablyfusio

ninhibitors

anti-inflammatory

Plantextractsmdash

Cinn

amom

umcassia

Cimicifuga

foetida


Decoctio

ns-m

odified

Dingchu

an

Liu-He-Tang

water

extracto

fLicorice

Prom

ising

butcon

clusiv

estudies

requ

ired

[158ndash164

]























6 Conclusions


Competing Interests


Acknowledgments



References




















































































































































































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology










4 Treatment








Table2Com

paris

onof

different

treatmentapp

roachesfor

RSV

Treatm

ent

Mechanism

Exam

ple

Remark

References

Antivira

ldrugs


nRS

V60

4Eff

ectiv

eagainstRS

Vbu

tadverse

effecto

ntheh

ost

[104]

[142ndash146]

Mutation

Ribavirin

vira

midinemerim

epod

ibInhibitoro

fino

sinem

onop

hosphate

dehydrogenase

Ribavirin

mycop

heno

latem

izoribine

Immun

ostim

ulatoryeffects

Ribavirin

Fusio

ninhibitors

Inhibitin

gfusio


Peptidemdash

HR121H

R212R

hoA

Chem

icalmdashBM

S-433771R

FI-641


ising

anti


ninhibitorshave

sidee

ffects

[147ndash150]

Nanop

articles

Inhibitin

gattachmenttocell

Silver

nano

particlesgold

nano

particles

Emerging

fieldcon

clusiv

estudies

requ

ired

[151152]

Antise

nsetherapy

RNAinterfe

rence

siRNA-ALN

-RSV

01Ph

osph

orod

iamidatem

orph

olinooligom

ersEff

ectiv

eand

safeA

LN-RSV

01completed

phaseIIb

clinicaltrails

[153ndash157]

Ethn

obotanicals

Prob

ablyfusio

ninhibitors

anti-inflammatory

Plantextractsmdash

Cinn

amom

umcassia

Cimicifuga

foetida


Decoctio

ns-m

odified

Dingchu

an

Liu-He-Tang

water

extracto

fLicorice

Prom

ising

butcon

clusiv

estudies

requ

ired

[158ndash164

]























6 Conclusions


Competing Interests


Acknowledgments



References




















































































































































































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology



4 Treatment








Table2Com

paris

onof

different

treatmentapp

roachesfor

RSV

Treatm

ent

Mechanism

Exam

ple

Remark

References

Antivira

ldrugs


nRS

V60

4Eff

ectiv

eagainstRS

Vbu

tadverse

effecto

ntheh

ost

[104]

[142ndash146]

Mutation

Ribavirin

vira

midinemerim

epod

ibInhibitoro

fino

sinem

onop

hosphate

dehydrogenase

Ribavirin

mycop

heno

latem

izoribine

Immun

ostim

ulatoryeffects

Ribavirin

Fusio

ninhibitors

Inhibitin

gfusio


Peptidemdash

HR121H

R212R

hoA

Chem

icalmdashBM

S-433771R

FI-641


ising

anti


ninhibitorshave

sidee

ffects

[147ndash150]

Nanop

articles

Inhibitin

gattachmenttocell

Silver

nano

particlesgold

nano

particles

Emerging

fieldcon

clusiv

estudies

requ

ired

[151152]

Antise

nsetherapy

RNAinterfe

rence

siRNA-ALN

-RSV

01Ph

osph

orod

iamidatem

orph

olinooligom

ersEff

ectiv

eand

safeA

LN-RSV

01completed

phaseIIb

clinicaltrails

[153ndash157]

Ethn

obotanicals

Prob

ablyfusio

ninhibitors

anti-inflammatory

Plantextractsmdash

Cinn

amom

umcassia

Cimicifuga

foetida


Decoctio

ns-m

odified

Dingchu

an

Liu-He-Tang

water

extracto

fLicorice

Prom

ising

butcon

clusiv

estudies

requ

ired

[158ndash164

]























6 Conclusions


Competing Interests


Acknowledgments



References




















































































































































































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


Table2Com

paris

onof

different

treatmentapp

roachesfor

RSV

Treatm

ent

Mechanism

Exam

ple

Remark

References

Antivira

ldrugs


nRS

V60

4Eff

ectiv

eagainstRS

Vbu

tadverse

effecto

ntheh

ost

[104]

[142ndash146]

Mutation

Ribavirin

vira

midinemerim

epod

ibInhibitoro

fino

sinem

onop

hosphate

dehydrogenase

Ribavirin

mycop

heno

latem

izoribine

Immun

ostim

ulatoryeffects

Ribavirin

Fusio

ninhibitors

Inhibitin

gfusio


Peptidemdash

HR121H

R212R

hoA

Chem

icalmdashBM

S-433771R

FI-641


ising

anti


ninhibitorshave

sidee

ffects

[147ndash150]

Nanop

articles

Inhibitin

gattachmenttocell

Silver

nano

particlesgold

nano

particles

Emerging

fieldcon

clusiv

estudies

requ

ired

[151152]

Antise

nsetherapy

RNAinterfe

rence

siRNA-ALN

-RSV

01Ph

osph

orod

iamidatem

orph

olinooligom

ersEff

ectiv

eand

safeA

LN-RSV

01completed

phaseIIb

clinicaltrails

[153ndash157]

Ethn

obotanicals

Prob

ablyfusio

ninhibitors

anti-inflammatory

Plantextractsmdash

Cinn

amom

umcassia

Cimicifuga

foetida


Decoctio

ns-m

odified

Dingchu

an

Liu-He-Tang

water

extracto

fLicorice

Prom

ising

butcon

clusiv

estudies

requ

ired

[158ndash164

]























6 Conclusions


Competing Interests


Acknowledgments



References




















































































































































































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology























6 Conclusions


Competing Interests


Acknowledgments



References




















































































































































































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology
















6 Conclusions


Competing Interests


Acknowledgments



References




















































































































































































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology








6 Conclusions


Competing Interests


Acknowledgments



References




















































































































































































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


References




















































































































































































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


















































































































































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


















































































































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

Documents

Review Article Recent Advances in Diagnosis, Prevention