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    Rapid Detection and Subtyping of Human Influenza A Viruses and Reassortants by Pyrosequencing

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    33
    Author
    Deng, Y-M; Caldwell, N; Barr, IG
    Date
    2011-08-19
    Source Title
    PLoS One
    Publisher
    PUBLIC LIBRARY SCIENCE
    University of Melbourne Author/s
    Barr, Ian
    Affiliation
    Microbiology and Immunology
    Metadata
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    Document Type
    Journal Article
    Citations
    Deng, Y. -M., Caldwell, N. & Barr, I. G. (2011). Rapid Detection and Subtyping of Human Influenza A Viruses and Reassortants by Pyrosequencing. PLOS ONE, 6 (8), https://doi.org/10.1371/journal.pone.0023400.
    Access Status
    Open Access
    URI
    http://hdl.handle.net/11343/259598
    DOI
    10.1371/journal.pone.0023400
    Abstract
    BACKGROUND: Given the continuing co-circulation of the 2009 H1N1 pandemic influenza A viruses with seasonal H3N2 viruses, rapid and reliable detection of newly emerging influenza reassortant viruses is important to enhance our influenza surveillance. METHODOLOGY/PRINCIPAL FINDINGS: A novel pyrosequencing assay was developed for the rapid identification and subtyping of potential human influenza A virus reassortants based on all eight gene segments of the virus. Except for HA and NA genes, one universal set of primers was used to amplify and subtype each of the six internal genes. With this method, all eight gene segments of 57 laboratory isolates and 17 original specimens of seasonal H1N1, H3N2 and 2009 H1N1 pandemic viruses were correctly matched with their corresponding subtypes. In addition, this method was shown to be capable of detecting reassortant viruses by correctly identifying the source of all 8 gene segments from three vaccine production reassortant viruses and three H1N2 viruses. CONCLUSIONS/SIGNIFICANCE: In summary, this pyrosequencing assay is a sensitive and specific procedure for screening large numbers of viruses for reassortment events amongst the commonly circulating human influenza A viruses, which is more rapid and cheaper than using conventional sequencing approaches.

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