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Detection of influenza A(H1N1)v virus by real-time RT-PCR

2009; European Centre for Disease Prevention and Control; Volume: 14; Issue: 36 Linguagem: Inglês

10.2807/ese.14.36.19329-en

1560-7917

Marcus Panning, Markus Eickmann, Olfert Landt, Masyar Monazahian, Stephan Ölschläger, Sigrid Baumgarte, Udo Reischl, Jürgen J. Wenzel, Hans Helmut Niller, Stephan Günther, B. Hollmann, Daniela Huzly, Jan Felix Drexler, Angelika Helmer, Stephan Becker, B. Matz, Anna Maria Eis‐Hübinger, Christian Drosten,

Virus-based gene therapy research

Influenza A(H1N1)v virus was first identified in April 2009. A novel real-time RT-PCR for influenza A(H1N1)v virus was set up ad hoc and validated following industry-standard criteria. The lower limit of detection of the assay was 384 copies of viral RNA per ml of viral transport medium (95% confidence interval: 273-876 RNA copies/ml). Specificity was 100% as assessed on a panel of reference samples including seasonal human influenza A virus H1N1 and H3N2, highly pathogenic avian influenza A virus H5N1 and porcine influenza A virus H1N1, H1N2 and H3N2 samples. The real-time RT-PCR assay for the influenza A matrix gene recommended in 2007 by the World Health Organization was modified to work under the same reaction conditions as the influenza A(H1N1)v virus-specific test. Both assays were equally sensitive. Clinical applicability of both assays was demonstrated by screening of almost 2,000 suspected influenza (H1N1)v specimens, which included samples from the first cases of pandemic H1N1 influenza imported to Germany. Measuring influenza A(H1N1)v virus concentrations in 144 laboratory-confirmed samples yielded a median of 4.6 log RNA copies/ml. The new methodology proved its principle and might assist public health laboratories in the upcoming influenza pandemic.