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


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2001-12

AUTHORS

E. Hoffmann, J. Stech, Y. Guan, R. G. Webster, D. R. Perez

ABSTRACT

Summary. 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. More... »

PAGES

2275-2289

Journal

TITLE

Archives of Virology

ISSUE

12

VOLUME

146

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1007/s007050170002

    DOI

    http://dx.doi.org/10.1007/s007050170002

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1004286290

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/11811679


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