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      Universal primer set for the full-length amplification of all influenza A viruses

      , , , ,
      Archives of Virology
      Springer Science and Business Media LLC

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          Abstract

          To systematically identify and analyze the 15 HA and 9 NA subtypes of influenza A virus, we need reliable, simple methods that not only characterize partial sequences but analyze the entire influenza A genome. We designed primers based on the fact that the 15 and 21 terminal segment specific nucleotides of the genomic viral RNA are conserved between all influenza A viruses and unique for each segment. The primers designed for each segment contain influenza virus specific nucleotides at their 3'-end and non-influenza virus nucleotides at the 5'-end. With this set of primers, we were able to amplify all eight segments of N1, N2, N4, N5, and N8 subtypes. For N3, N6, N7, and N9 subtypes, the segment specific sequences of the neuraminidase genes are different. Therefore, we optimized the primer design to allow the amplification of those neuraminidase genes as well. The resultant primer set is suitable for all influenza A viruses to generate full-length cDNAs, to subtype viruses, to sequence their DNA, and to construct expression plasmids for reverse genetics systems.

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          Author and article information

          Journal
          705
          Archives of Virology
          Archives of Virology
          Springer Science and Business Media LLC
          0304-8608
          1432-8798
          December 1 2001
          December 1 2001
          : 146
          : 12
          : 2275-2289
          Article
          10.1007/s007050170002
          11811679
          3c2916e7-9f3a-45a1-8522-124fd94641a9
          © 2001

          http://www.springer.com/tdm

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