使用亚型特异性引物对全长测序所有 9 种亚型的流感 A 病毒的神经氨酸酶基因。
Full length sequencing of all nine subtypes of the neuraminidase gene of influenza A viruses using subtype specific primer sets.
机构信息
Department of Veterinary Population Medicine, College of Veterinary Medicine, University of Minnesota, 1333 Gortner Avenue, Saint Paul, MN 55108, United States.
出版信息
J Virol Methods. 2010 Apr;165(1):116-20. doi: 10.1016/j.jviromet.2010.01.002. Epub 2010 Jan 28.
Abstract
An RT-PCR based method was developed using subtype specific overlapping primers to obtain full length amplification of neuraminidase (NA) gene from all subtypes (N1-N9) of influenza A viruses. This method was validated using reference strains of avian influenza viruses (AIV) (N1-N9), human influenza viruses (N1 and N2), and swine influenza viruses (N1-N3). Amplification of the NA gene was obtained with all viruses tested. Additionally, 200 field isolates of AIV from wild birds were tested by this method and the NA gene was amplified in all isolates. The NA subtype of all 200 isolates was determined by further sequencing of the amplified NA genes and all sequences were submitted to GenBank. The method described in this paper can be used to determine subtype of influenza isolates as well as their evolution and mutations if any, in the NA gene.
摘要
本研究开发了一种基于 RT-PCR 的方法，使用亚型特异性重叠引物对所有亚型（N1-N9）的流感 A 病毒的神经氨酸酶（NA）基因进行全长扩增。该方法使用禽流感病毒（AIV）（N1-N9）、人流感病毒（N1 和 N2）和猪流感病毒（N1-N3）的参考株进行了验证。用所有测试的病毒都获得了 NA 基因的扩增。此外，用该方法检测了 200 株来自野生鸟类的 AIV 分离株，所有分离株均扩增了 NA 基因。通过进一步对扩增的 NA 基因进行测序确定了所有 200 个分离株的 NA 亚型，并将所有序列提交给 GenBank。本文所述的方法可用于确定流感分离株的亚型及其在 NA 基因中的进化和突变（如果有）。
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