Single-Reaction Genomic Amplification Accelerates Sequencing and Vaccine Production for Classical and Swine Origin Human Influenza A Viruses

被引：495

作者：

Zhou, Bin ^{[1
,2
]}

Donnelly, Matthew E. ^{[1
]}

Scholes, Derek T. ^{[1
]}

George, Kirsten St. ^{[1
]}

Hatta, Masato ^{[3
]}

Kawaoka, Yoshihiro ^{[3
,4
]}

Wentworth, David E. ^{[1
,2
]}

机构：

[1] New York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USA

[2] SUNY Albany, Sch Publ Hlth, Dept Biomed Sci, Albany, NY 12201 USA

[3] Univ Wisconsin, Sch Vet Med, Dept Pathobiol Sci, Madison, WI 53706 USA

[4] Univ Tokyo, Inst Med Sci, Dept Microbiol & Immunol, Div Virol, Tokyo 1088639, Japan

来源：

JOURNAL OF VIROLOGY | 2009年 / 83卷 / 19期

基金：

日本科学技术振兴机构;

关键词：

GENERATION; TRANSCRIPTION; POLYMERASE; RNA; ATTENUATION; SYSTEM; GENES; H1N1; VRNA;

D O I：

10.1128/JVI.01109-09

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

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).

引用

页码：10309 / 10313

页数：5

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