鸡传染性支气管炎病毒豫北株的分离与鉴定

目的]分离与鉴定鸡传染性支气管炎病毒。方法]从豫北某蛋鸡场疑似鸡传染性支气管炎的病鸡中采集气管分泌物、肾脏等组织分离病毒,进行血凝试验、NDV干扰试验,采用反转录-聚合酶链式反应(RT-PCR)技术对HN07的纤突蛋白S1基因进行扩增、克隆和序列测定。结果]结果表明,分离到1株病毒,经连续鸡胚传代后,出现侏儒胚和蜷缩胚;血凝试验中,尿囊液无血凝性,而经胰酶处理后有血凝性;NDV干扰试验中,分离的尿囊液明显干扰NDV的增殖;病毒回归试验中病毒可引发接种鸡37.5%死亡;RT-PCR检测试验中,扩增到了约为1.7 kb的目的片段。DNAstar软件分析表明,测定基因具有IBVS1基因的共有分子特征,而且分离株HN07和HD株S1的同源性最高,核苷酸同源性为93.6%。结论]分离的病毒株HN07为鸡传染性支气管炎病毒。

Isolation and Identification of Avian Infectious Bronchitis Virus in North Henan

Objective]The purpose was to isolate and identify of avian infectious bronchitis virus.Method] The strain of virus was isolated from the kidney and liver tissue of dead chook in a big farm of North Henan to progress congealing blood and NDV interference experiment.S1 gene of HN07 was amplified,cloned and determined by RT-PCR.Result]The results indicated that a strain of virus was isolated from the kidney and liver tissue of dead chook,but it appeared stunted and curled embryos after continuous passage era of chicken embryo.Chicken red blood cells were agglutinated just with the isolates treated by trypsase.The isolate interfered remarkably in multiplication of LaSota strain of newcastle disease virus in the viral interference test.The experimental chickens infected with the isolate resulted 37.5% mortality rate.An objective fraction of 1.7 kb was amplified in RT-PCR identification test.In addition,S1 gene of HN07 was amplified by RT-PCR and sequenced.Sequence analysis showed that the nucleotide sequence was homogenous(homology rate of 93.6%) between HN07 and S1 gene of strain HD.Furthermore the phylogenetic trees analysis revealed that the isolates HN07 was an IBV strain.Conclusion]This study suggested that the isolated IBV strain of HN07 is avian infectious bronchitis.