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J Virol. 2009 Oct;83(19):10309-13. doi: 10.1128/JVI.01109-09. Epub 2009 Jul 15.

Single-reaction genomic amplification accelerates sequencing and vaccine production for classical and Swine origin human influenza a viruses.

Author information

1
Wadsworth Center, New York State Department of Health, Albany, New York 12201, USA.

Abstract

Pandemic influenza A viruses that emerge from animal reservoirs are inevitable. Therefore, rapid genomic analysis and creation of vaccines are vital. We developed a multisegment reverse transcription-PCR (M-RTPCR) approach that simultaneously amplifies eight genomic RNA segments, irrespective of virus subtype. M-RTPCR amplicons can be used for high-throughput sequencing and/or cloned into modified reverse-genetics plasmids via regions of sequence identity. We used these procedures to rescue a contemporary H3N2 virus and a swine origin H1N1 virus directly from human swab specimens. Together, M-RTPCR and the modified reverse-genetics plasmids that we designed streamline the creation of vaccine seed stocks (9 to 12 days).

PMID:
19605485
PMCID:
PMC2748056
DOI:
10.1128/JVI.01109-09
[Indexed for MEDLINE]
Free PMC Article

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