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目的了解2009年深圳市H3亚型流感病毒系统进化及分子变异情况。方法用狗肾传代细胞(MDCK)进行流感病毒分离,收获病毒后提取病毒RNA,进行逆转录-聚合酶链式反应(RT-PCR),扩增产物经琼脂糖凝胶电泳鉴定后送大连宝生物公司纯化及测序,测序结果用Clustal W2软件和MEGA 3.1软件进行序列分析。结果 2009年的深圳毒株与同期国际疫苗株“A/Brisbane/10/2007”相比差异较大,在HA1区段共发生了7个氨基酸位点的变异:62位的E突变成了K;抗原决定簇A的144位点由N变成了K;抗原决定簇B的158位点由K变成了N;173位的K变成了Q;抗原决定簇B的189位点由N变成了K;194位的P变成了L;213位的V突变成了A。值得注意的是N144K突变导致了144位糖基化位点的消失,这些突变可能已经造成了病毒抗原性的改变。结论 2009年深圳市流行的H3亚型流感病毒很可能是一个新的优势流行株。
Objective To understand the evolution and molecular variation of the H3 subtype influenza virus in Shenzhen in 2009. Methods MDCK was used to isolate influenza virus. The viral RNA was harvested after being harvested for reverse transcription - polymerase chain reaction (RT - PCR). The amplified product was identified by agarose gel electrophoresis and sent to Dalian Bao Bio-company purification and sequencing, sequencing results using Clustal W2 software and MEGA 3.1 software sequence analysis. Results The Shenzhen strain in 2009 was significantly different from the international vaccine strain “A / Brisbane / 10/2007” in the same period. A total of seven amino acid mutations were observed in the HA1 region: the E mutation at position 62 Became K; the epitope 144 of A was changed from N to K; the 158 site of antigenic determinant B was changed from K to N; the K at position 173 became Q; the 189 of epitope B The point changes from N to K; the 194th P becomes L; and the 213th V changes to A It is noteworthy that the N144K mutation led to the disappearance of the 144 glycosylation site and that these mutations may have caused a change in viral antigenicity. Conclusion The prevalence of H3 subtype influenza virus in Shenzhen in 2009 is likely to be a new prevalent epidemic strain.