[摘要] 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。
[关键词] 甲型流感病毒; RT - PCR; 克隆
Isolation and identification of H1N1 influenza Avirus in birds
[Abstract] 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 - PCR1.
[ Key words] Influenza A virus; RT - PCR; Clone
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