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InfluenzaA H1N1: Shell vial culture vs. PCR

Abstract number: R2335

Gonzalez Barbera E.M., Acosta Boga B., Fagundez Machain G., Molina Moreno J.M., Gobernado Serrano M.

In the beginning of the H1N1 Influenzavirus pandemia, we started working with PCR, culture and antigen detection (the last was refused due to its low sensibility), but culture was performed in certain circumstances like immunocompromised or admitted in Intensive Care Units patients.

Objectives: To compare shell-vial culture and PCR for Influenza AH1N1 detection considering PCR as "gold standard".

Methods: 214 samples were processed during the period April-November 2009 by PCR [AgPath-ID™One-Step RT-PCR Kit (Ambion Applied Biosystem), artus® Influenza/H1 LC/RG RT-PCR-Kit (Qiagen)] and cultured in MDCK cell line shell-vial (Vircell™) with Trypsine, incubated at 35–37°C for 24 or 48 hours, and stained with monoclonal antibody against respiratory viruses, including Influenza A (Chemicon™).

Results: See Table 1.

We also observed that 24 hours of incubation are enough and correlates with 50–75% cell monolayer infection (detected by monoclonal antibody stain).

Conclusion: MDCK cell line is an adequate cell line for Influenzavirus H1N1 isolation and Influenza A monoclonal antibody stain identifies it correctly in a pandemic situation (without distinguish between virus subtypes).

We consider that virus isolation is necessary for later studies. Furthermore, we may identify other viruses when we are not looking for them on purpose or in samples not considered at first.

Table 1. Culture and PCR comparison results

 Culture +Culture -Total
PCR +10232134
PCR -27880
Total104110214
Shell-vial culture sensibility    76.12%
Shell-vial culture specificity    97.5%
Shell-vial culture positive predictive value    98.08%

Session Details

Date: 10/04/2010
Time: 00:00-00:00
Session name: Abstracts 20th European Congress of Clinical Microbiology and Infectious Diseases
Subject:
Location: Vienna, Austria, 10 - 13 April 2010
Presentation type:
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