Giuseppe
Emeritus
J Virol. 2009 Jul 15. [Epub ahead of print]
Single Reaction Genomic Amplification Accelerates Sequencing and Vaccine Production for Classical and Swine Origin Human Influenza A Viruses.
Zhou B, Donnelly ME, Scholes DT, St George K, Hatta M, Kawaoka Y, Wentworth DE. Wadsworth Center, New York State Department of Health, and Department of Biomedical Sciences, School of Public Health, State University of New York, Albany, New York 12201, USA; and Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI 53706, USA.
Pandemic influenza A viruses that emerge from animal reservoirs are inevitable. Therefore, rapid genomic analysis, and creation of vaccines are vital. We developed a "multi-segment RT-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 we designed streamline the creation of vaccine seed stocks (9-12 days).
PMID: 19605485 [PubMed - as supplied by publishe
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Single Reaction Genomic Amplification Accelerates Sequencing and Vaccine Production for Classical and Swine Origin Human Influenza A Viruses.
Zhou B, Donnelly ME, Scholes DT, St George K, Hatta M, Kawaoka Y, Wentworth DE. Wadsworth Center, New York State Department of Health, and Department of Biomedical Sciences, School of Public Health, State University of New York, Albany, New York 12201, USA; and Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI 53706, USA.
Pandemic influenza A viruses that emerge from animal reservoirs are inevitable. Therefore, rapid genomic analysis, and creation of vaccines are vital. We developed a "multi-segment RT-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 we designed streamline the creation of vaccine seed stocks (9-12 days).
PMID: 19605485 [PubMed - as supplied by publishe
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