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S.Tsatsral, NCCD, Department of Virology

Human rhinoviruses (HRVs) are small, single-stranded, positive-sense RNA viruses that are nowclassified within the genus Enterovirus in the familyPicarnoviridae.

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*The first HRV was discovered in 1953 and by now HRVswere classified into approximately 150 genotypes ofHRV-A, B and C groups .

*HRVs are the most frequent causes of acute upperrespiratory tract infections (RTIs). HRV illnesses arecommon in all age groups, they occur throughout theyear and they are present world-wide. In longitudinalfamily studies the overall HRV isolation rate in personswith RTIs has varied from 6.1% to 23.3.

We have studied 2515 Nasopharyngeal swabswhich were negative for influenza viruses. These sampleswere collected from November 2008 to March 2013 frompatients with acute RTIs and were kept at the NationalCenter of Communicable Diseases (NCCD).

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I – Rhinoviruses detection in clinical samples

II – Rhinovirus genotyping (Sequence)

Rhinoviruses detection in clinical samples

*RNA extraction

(ExiPrep™ Viral DNA/RNA kit, Bioneer, Korea)

*Rhinoviruses detection for real time PCR

(“FTD respiratory pathogen 21” kit, Fast track diagnostics, Luxembourg)

Rhinovirus genotyping (Sequence)

*PCR amplification- VP4/VP2

- 5’UTR

*Clean PCR product (Gel purification)

*Ligate PCR product (Cloning PCR products, pGEM-T Easy Vector, Promega)

*Transform bacteria (E-Coli, DH5α competent cell)

*Plate bacteria (LB/amp/IPTG/X-Gal plate, blue and white screening)

*Analyze sequence (ABI 3130xl genetic analyzer, M13 vector primer)

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Other89%

n=251611.4% (286/2516) Rhinovirus

positive 11.4% Outpatient

53%

Hospital patient47%

Adenovirus7%

Coronavirus19%

Enterovirus7%

Influenza virus4%

Bocavirus26%

Metapneumovirus4%

Parainfluenza virus14%

RSV19%

n=7426% (74/286)

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11.7

13.3

11.4

9.4

7.7

0.0

2.0

4.0

6.0

8.0

10.0

12.0

14.0

0-5 6-10 11-16 17-46 ≥47

Det

ecte

d p

erce

nta

ge

Age groups

0

5

10

15

20

25

30

Jan

Feb

Mar

Apr

May

Jun

Jul

Aug

Sep

Oct

Nov

Dec

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0

20

40

60

80

100

120

HRVA HRVB HRVC

46

7

40

62

9

54

Hospital patients Outpatients

Median age 2

Median age 3

Median age 7

*HRV was detected throughout the year and in all age groups.

*From all genotypes of HRV genotypes HRVA and HRVC were more frequently detected than HRVB genotype.

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