从鸟类中分离到的H1N1流感病毒株的鉴定

Isolation and identification of H1N1 influenza A virus in birds

目的:对采集新鲜鸟粪便中甲型流感病毒进行检测、分离和鉴定。方法:采用鸡胚进行病毒分离,采用血凝试验初步确定病毒效价,用逆转录聚合酶链反应法(RT-PCR)扩增分离株的部分片段后进行克隆和测序,然后进行进化树分析。结果:从采集的80份新鲜鸟粪便中分离到一株低致病性流感病毒,其编号为WH-1,测出的部分NP基因序列经Blast比较,与A/Canterbury/106/2004/H1N1的同源性最高,核苷酸为99%,氨基酸为100%,与GenBank中收录的AIV其它病毒株NP基因相应片段的同源性在96%以上。结论:分离得到WH-1为一株新的甲型流感分离毒株,该分离株可以用RT-PCR方法进行检测。

Objective：To isolate and detect influenza A virus from fresh bird excrement and identify its genotype. Mothods： Cock embryo was used to isolate influenza viruses, and the virus titer was determined through hemagglutination test. The gene was amplified by RT - PCR technique from the isolated strain and sequenced, and the gene phylogenetic tree was analyzed. Results： A positive strain of avian influenza virus was isolated from 80 fresh bird excrement, named WH - 1, and the nucleotide and amino acids sequence showed the highest homology with A/Canterbury/106/2004/H1N1 stain, and identities were 99% and 100%, respectively. Conclusion：The acquried WH - 1 is a new influenza A virus isolated strain and it can be detected by RT- PCR.