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

Content Provider	Springer Nature Link
Author	Hoffmann, E. Stech, J. Guan, Y. Webster, R. G. Perez, D. R.
Copyright Year	2001
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.
Starting Page	2275
Ending Page	2289
Page Count	15
File Format	PDF
ISSN	03048608
Journal	Archives of Virology
Volume Number	146
Issue Number	12
e-ISSN	14328798
Language	English
Publisher	Springer-Verlag
Publisher Date	2001-12-01
Publisher Place	Wien
Access Restriction	One Nation One Subscription (ONOS)
Content Type	Text
Resource Type	Article
Subject	Virology Medicine

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