期刊
JOURNAL OF VIROLOGY
卷 83, 期 19, 页码 10309-10313出版社
AMER SOC MICROBIOLOGY
DOI: 10.1128/JVI.01109-09
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资金
- NIH/NIAID [R21AI 057941-02]
- New York State Department of Health
- National Institute of Allergy and Infectious Diseases Public Health Service
- Ministry of Health, and by ERATO ( Japan Science and Technology Agency)
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).
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