NEW TECHNOLOGIES IN DIAGNOSIS OF TUBERCULOSIS by Dr.T.V.Rao MD

8/20/2019 NEW TECHNOLOGIES IN DIAGNOSIS OF TUBERCULOSIS by Dr.T.V.Rao MD

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NEW TECHNOLOGIES INDIAGNOSIS OF

TUBERCULOSISDr.T.V.Rao MD

3/5/16 Dr.T.V.Rao MD



A Tribute to Robert KocTe M!"ter! o# Tubercu$o"i" Co%ti




a'( e" re)u re o'icrobio$o+ica$ *ia+%o

o# TB






A*,a%ci%+ Micro"co(! 1iLi+t E'itti%+ Dio*e" LED

*e,e$o('e%t o$i+t e'itti%+*io*e" LED/- 1

te a((ro(riat0uore"ce%t $i+out(ut #or FM $o1 (o1erco%"u'(tio%-

$e* to te*e,e$o('e%t o"i'($e- robu"tFM 'icro"co(e

re)uiri%+ 'i%i3/5/16 Dr.T.V.Rao MD



Micro"co(! a%* Cu$turi%Sti$$ #ea"ib$e A$ter%ati,e

• Micro"co(ice3a'i%atio% i""uitab$e #or$aboratorie" at(eri(era$ a%*

i+er $e,e$" a%* itca% be *o%e "a#e$!u%*er 'i%i'a$bio"a#et!co%*itio%".

M!cobacteria$3/5/16 Dr.T.V.Rao MD



A%tibiotic "e%"iti,it! te"ti%+Co%,e%tio%a$ 'eto*" +etti%

*ate*




WHO Reco''e%*i%+ t

• The WHO hasrecommendedrolling i o! as analernai"e o #Ms in

reso!rce$limiedseings% &ased ons!dies ha ha"esho'n com(ara&le(er)ormance o) #Mand sandard *M3/5/16 Dr.T.V.Rao MD



SCIENCE IS MOVING

FROM BIOLOG5 TOMICROBIOLOG5




Mo$ecu$ar Bio$o+! a*,a%c1it Nuc$eic aci* ba"e* te"

•

NAAT" are 'o$ecu$ar"!"te'" tat ca%*etect "'a$$ )ua%titie"o# +e%etic 'ateria$DNA or RNA/ #ro''icroor+a%i"'" "uc

a" M!cobacteriu'tubercu$o"i". 6CR i" te'o"t co''o% a'o%+te ,ariet! o# NAAT".NAAT" are a,ai$ab$e a"

co''ercia$ 2it" a%* i%4ou"e te"t" a%* are3/5/16 Dr.T.V.Rao MD



Mo$ecu$ar Bio$o+! a*,a%ce" 1Nuc$eic aci* ba"e* te"t"

•I%4ou"e 6CR te"t"are 1i*e$! u"e* i%te *e,e$o(i%+cou%trie" becau"ete"e te"t" are $e""

e3(e%"i,e ta%co''ercia$ 2it". Te$i%e (robe a""a!"L6A/ are "uitab$eo%$! #or %atio%a$ or

re+io%a$ $e,e$3/5/16 Dr.T.V.Rao MD



Wat are Mo$ecu$ar te"

• n micro&iolog+% heerm molec!lar esis con"enionall+!sed o descri&e

ess ha deec(ahogen n!cleicacid 0D or R.ll molec!lardiagnosic essha"e hree sages




Ge%ot!(ic 'eto*•Molec!lar echni!es are aimed a he nacid o) he m+co&aceri!m as he anal+Ri&osomalrR is !se)!l geneic arge )idenicaion o) organisms% since i o)enconains s(esic se!ences and is (rese

he cells and media in high !ani+ d!egro'h o) he m+co&aceria. There are "a((licaions o) molec!lar echni!es )or deecion and idenicaion o) MT8.




E3tractio% o# Nuc$eic ac

• The eracion (sho!ld &e carrieda conainmen lela&oraories and

(ro"ide inaci"aorganisms since :ro!( 3 s(ecies &e (resen




A'($i7catio%• The n!cleic acid inhe sam(le orc!l!re is am(liedso ha i can &e

deeced.;ol+merase chainreacion 0;<R is aneam(le o) amehod )or

am(licaion% &! a3/5/16 Dr.T.V.Rao MD



6o$!'era"e Cai% React

•

;<R is he common )ormao) n!cleic acidam(licaion ess 0T=ohemehodologiesincl!deligase chain reacion% sraindis(lacemen am(licaion%

loo($mediaed isohermalam(li caion 0#M; andranscri(ion mediaedam(licaion. Morerecenl+% real$ime




RT 6CR•RT ;<Rechnologies &asedon >!orescen$(ro&e deecion or

meling$c!r"eanal+sis ha"e &eende"elo(ed


M t * #



Detectio%• Meto*" o#*etectio% o# ta'($i7e* %uc$aci* a$"o ,ar!

i%c$u*e te u"e$ectro(ore"(robe a""a!" oti'e *etectio%

De(e%*i%+ o% #or'at- "o'e t'a! ta2e t1o #or re"u$t" to ba,ai$ab$e 1eroter" 'a ro




Di8ere%t t!(e" o# 'o$ecu$te"ti%+ tec%o$o+! are

a,ai$ab$e #or TB




Ru$e" o# te LabK%o1 be#ore be$ie,e e,er!t

•No $ab te"t i" (er#ect

• Do %ot or*er a $ab te"t i# !ou %ot rea*! to *ea$1it te re"u

•

Treat te (atie%t- %ot te $abte"t) For TB9Tere i" a $ot 1e%ee* to $ear% about

•DST a%* 'o$ecu$ar *etectio% o

*ru+ re"i"ta%ce3/5/16 Dr.T.V.Rao MD



NAAT #or TB Dia+%o• Beco'i%+ "ta%*ar* o# (ractice

• CANNOT re($ace c$i%ica$ :u*+'e%t or be re$ie* te

• ONL5 +ui*e #or tera(! or i"o$atio% (ractice" $ta%(er#ect "e%"iti,it! a%* "(eci7cit!/

Se%"iti,it!;<=> #or AFB "'ear4(o"iti,e TB(atie%t"==?@=> o# AFB "'ear4%e+ati,e- cu$ture4(o"iti,e TB 6atie%t"

6er#or'a%ce i'(ro,e" 1it i%crea"e* c$i%ica$ "uo#TB

6ote%tia$ be%e7t" ? i'(act o% tera(!- (ub$ic3/5/16 Dr.T.V.Rao MD

o ecu ar e o " e



o ecu ar e o " eDetectio% Dru+

Re"i"ta%ce too•

These molec!lar echni!es also aimdeecing resisance genes. ?am(leincl!des= D (ro&e and D se!eo) MT8 gene s!ch as caalase 0@a:

R (ol+merase0r(o8. M!aions inhese genes ha"e &een associaed 'resisance o isonia9id and ri)am(icires(eci"el+.




Mo$ecu$ar Te"t" are Ra(i*e3(e%"i,e too

•

The !sing o) molec!lar (rimers in real$ imereacion can diAereniae &e'een he (reshe 'ild$ +(e se!ence and m!aed se!eassociaed 'ih dr!g resisance. Molec!lar are ra(id 0'ihin )e' ho!rs% highl+ sensii"e

s(ecic% &! e(ensi"e% re!ires e(erise ama+ no diAereniae aci"e in)ecion as Da dead organism d!ring ani&ioic reamen&e deeced and am(lied &+ ;<R




Li%e4(robe a""a!•

#ine$(ro&e assa+s andB;?RT MT8/R* #ine(ro&e assa+s 0#;sare ac!al molec!laress. Three main #;s

)or he ra(id diagnosiso) T8 and/or ra(iddeecion o) RM;resisance andMDR$/BDR$T8 are

c!rrenl+ a"aila&le on3/5/16 Dr.T.V.Rao MD

Li%e (robe a""a!" L6A"/ #o



Li%e (robe a""a!" L6A"/ #otubercu$o"i" 1ere e%*or"e* b!

WHO i% •

#ine (ro&e assa+s0#;s )or !&erc!losis'ere endorsed &+ heWHO in 2, )ormolec!lar deecion o)

dr!g resisance )romsmear$(osii"e(aiens a ris@ o) M!liDr!g Resisan$

T!&erc!losis 0MDRT8

• T'o commercial #;s3/5/16 Dr.T.V.Rao MD



Li%e 6robe A""a!" #or tubercu• #ine ;ro&e ssa+s )or!&erc!losis !ses;<R/h+&ridi9aion ec

ideni)+ mem&ers o) M+co&aceri!m T!&er'hile sim!laneo!sl+ideni)+ing dr!g$resissrains &+ deecing common single n!cle

(ol+mor(horisms 0Cassociaed 'ih resisMea$anal+ses ha"e sha #;s are highl+ a)or he deecion o) dr!g resisance% es(e

smear$(osii"e s(!!m3/5/16 Dr.T.V.Rao MD

Wor$* Hea$t Or+a%iatio% W



Wor$* Hea$t Or+a%iatio% Wreco''e%*" te u"e o# (e

MTBRIF•

Wor$* Hea$tOr+a%iatio% WHO/reco''e%*" te u"eo# (ert MTBRIFa""a! #or ra(i*

*ia+%o"i" o#tubercu$o"i" TB/ a%**etectio% o#ri#a'(ici% re"i"ta%ce.Ti" "!"te'atic

re,ie1 1a" *o%e to3/5/16 Dr.T.V.Rao MD



WHO E%*or"e" Ne1 Te"ti%Meto* i%

n earl+ 211% WHO endorsed a no"el% ra(id% molec!lar diagnosic es% he B(er MT8/R*

sim!laneo!sl+ deec T8 and ri)am(icin resisMT8/R* (ro"ides res!l 'ihin 2 h% h!s he (

sared 'ih (ro(er reamen e"en on he sam




Ge%oT!(eMTBDRMTBDR($u"Ge%o4T!(e

• Ge%oT!(eMTBDRMTBDR($u" a%* GeMTBDR"$ bot Hai%Li#e"cie%ce- Ger'a""a!" are ba"e* o% te tar+ete* a'(o# "(eci7c #ra+'e%t" o# te MTB +e%o!bri*i"atio% o# 6CR (ro*uct" to o$i+o

(robe" i''obi$i"e* o% 'e'bra%e"


N I ti M t *



Ne1er I%%o,ati,e Meto*Dia+%o"i" o# Tubercu$o"

• INNO4Li6A Ri# TB *etect" o%$! RM6re"i"ta%ce-

• Ge%oT!(e MTBDRMTBDR ($u" *etect"RM6 a%* INH re"i"ta%ce- a%* Ge%oT!(

• MTBDR"$*etect" re"i"ta%ce to

0uoro)ui%o$o%e"- i%:ectab$e "eco%*4$i*ru+" a%* eta'buto$.

• Te"e te"t" are *e"i+%e* #or *etectioMTB i"o$ate" i% re"(irator! "(eci'e%"(ert


$ $ *



Mo$ecu$ar Meto*" are %orouti%e$! U"e*

•

:eno+(icmehods are noro!inel+ !sed inhem+co&aceri!mla&oraor+= he+are esseniall+

)or research3/5/16 Dr.T.V.Rao MD



Ma%! Mo$ecu$ar Meto*" i% oJ

• M!li(le commercial essare a"aila&le and ne'ess are also &eingde"elo(ed. Comela&oraories ha"ede"elo(ed local in$

ho!seE ess. The choiceo) es de(ends on clinicalneed and local la&oraor+)acors s!ch as n!m&erso) s(ecimens recei"ed &+he la&oraor+% access omolec!lar la)orms and3/5/16 Dr.T.V.Rao MD



Li%e (robe a""a!

• Li%e (robe a""a!" a%* te Ce(ei* MTi" ru% o% te Ge%e(ert ($at#or' are "!"te'" tat are reco''e%*e* b! tere"ource (oor cou%trie" ? 1ere 7%a%a,ai$ab$e #or te co"t o# te"t". I% *e,eti" #or'at o# te"t i" co%,e%ie%t but %

or re$iab$e ta% oter te"t "!"te'" a%e3(e%"i,e.


i i i * C i* i



Li'itatio%" a%* Co%"i*eratioo% Mo$ecu$ar Meto*"

• Se%"iti,it!

• Re*uce* #or "'ear %e+ati,e "(eci'e%" a%* "o'e

• "(eci'e% t!(e"&

• Do !ou 1a%t to ru$e i% or ru$e out&

• 6$at#or' *e(e%*e%t

• ) S(eci7cit!

• 6$at#or' *e(e%*e%t• ) Doe" %ot re($ace %ee* #or cu$ture

• Cu$ture "ti$$ %ee*e* #or co%,e%tio%a$ DST- +e%ot!(i%+

• ) A'($ico% cro"" co%ta'i%atio% i% o(e% "!"te'"

• ) Co"t a%* "u"tai%abi$it!

• E3(e%"e ca% $i'it uti$iatio%3/5/16 Dr.T.V.Rao MD



Atte%tio% o# Vie1er"• I a' ta%2#u$ to 'a%! i% te 1or$* 1o 'a*e 'e to acie,e '! *e"ire* +oaI tou+t- a,i%+ ; 4= 'i$$io% ea$t (ro#e""io%a$" "are a%* uti$ie '! 2%o

be%e7t o# 'a%2i%*- To*a! I 1i" to be #ree$a%cer to te 1or$* to create i%teMe*ica$- C$i%ica$ a%* Dia+%o"tic Microbio$o+! 1it 'ore e'(a"i" o% I%#ecta%* Ho"(ita$ a""ociate* I%#ectio% 1i" to be !our (art%er i% e*ucati%+ 'a%!2%o1 1e$$ te i'(orta%ce o# I%#ectiou" *i"ea"e"

• 5ou ca% ,i"it 'a%! 1eb "ite" o# 'i%e

• 111.'e*'icrobe".co'

• 111."$i*e"are.co'

• 111.autour"trea'.co'

•

111-"crib*.co'• Be a #rie%* o% Faceboo2 1it tu''a$a(a$$i ,e%2ate"1ararao acce""

• Rao" Microbio$o+!

• Rao" I%#ectio% care

• Microbio$o+! co%%ecte* Tra,a%core Me*ica$ Co$$e+e

• For a%! a""i"ta%ce o% INFECTION REALTED ISSUES CONTACT ME AT *octort,raoP+'a

•Mob Q< @=3/5/16 Dr.T.V.Rao MD

http://www.medmicrobes.com/

http://www.slidehsare.com/

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mailto:[email protected]

mailto:[email protected]

http://www.authourstream.com/

http://www.slidehsare.com/

http://www.medmicrobes.com/



•6ro+ra' Create* b! Dr.T.V.Rao M

Be%e7t o# Me*ica$ a%* 6ara'e*6ro#e""io%a$" i% te De,e$o(i%+ WCreate* #ro' Wor$* Wi*e Re"ou

•E'ai$

•*octort,raoP+'ai$.co'


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NEW TECHNOLOGIES IN DIAGNOSIS OF TUBERCULOSIS by Dr.T.V.Rao MD