Blog
About

  • Record: found
  • Abstract: found
  • Article: not found

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

Archives of Virology

Neuraminidase, Humans, Child, Reverse Transcriptase Polymerase Chain Reaction, Oligonucleotides, Child, Preschool, Gene Amplification, genetics, Nucleic Acid Amplification Techniques, Hemagglutinin Glycoproteins, Influenza Virus, Influenza A virus, DNA Primers, classification, Influenza, Human, virology

Read this article at

ScienceOpenPubMed
Bookmark
      There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

      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.

      Related collections

      Author and article information

      Journal
      11811679

      Comments

      Comment on this article