A comprehensive systematic approach to identification of influenza A virus genotype using RT-PCR and RFLP

作者: Daniel P Offringa , Victoria Tyson-Medlock , Zhiping Ye , Roland A Levandowski

DOI: 10.1016/S0166-0934(00)00180-4

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摘要: Amplification of influenza A virus gene segments by reverse transcription-polymerase chain reaction (RT-PCR) can be combined with enzymatic digestion to reveal unique restriction fragment length polymorphisms specific for H1N1 and H3N2 subtype viruses. We have used the method provide a rapid, reproducible identification genotype high-growth reassortants derived from A/Puerto Rico/8/34 (PR8). Digestion amplified wild-type viruses, PR8 at sites either strain or provided positive, unambiguous origin each internal genes, distinguished genes both strains those PR8. This has also permitted us quickly confirm that reassorting occurred optimize selection reassortant clones maximum number genes. Since detect 1-10% second in mixed population, samples containing more than one viral assess purity viruses manufacturing vaccines.

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