牛传染性鼻气管炎病毒的分离鉴定及gD基因序列分析

目的]从疑似牛传染性鼻气管炎病毒感染的牛鼻腔棉拭子中分离出1株病毒,对其进行鉴定和序列分析。方法]从疑似牛传染性鼻气管炎病毒感染的牛鼻腔棉拭子中分离到1株病毒,将其命名为NM株。通过PCR、MDBK细胞病变观察、中和试验、动物回归试验、gD基因序列分析对其进行鉴定。结果]确定该分离株为牛传染性鼻气管炎病毒强毒株。NM株病毒能使MDBK细胞产生牛传染性鼻气管炎病毒典型性细胞病变。Reed-Muench法测定NM株病毒的TCID50为10-6.0/ml。NM株病毒与IBRV阳性血清发生中和反应,而与IBRV阴性血清不发生中和反应。NM株病毒能使8月龄IBRV抗体阴性牛致病,表现出牛传染性鼻气管炎的典型临床症状。gD基因序列鉴定结果表明NM株病毒与IBRV K22株的同源性最高。结论]该研究可为IBR诊断试剂与疫苗的研制奠定基础。

Isolation, Identification and Sequence Analysis of gD Gene of Infectious Bovine Rhinotracheitis Virus

Objective] The research aimed to isolate a strain of virus from nasal swab samples of cattle which might be infected by infectious bovine rhinotracheitis（IBR） and make the identification and sequencing analysis.Method] A strain of virus was isolated from nasal swab samples of cattle which might be infected by infectious bovine rhinotracheitis（IBR） and it was named as NM strain.The isolated strain was identified by PCR,MDBK cell lesion observation,neutralization test,animal regression test and gD gene sequence analysis.Result] The isolated strain was confirmed as virulent strain of infectious bovine rhinotracheitis virus.NM strain made MDBK cell to produce typical cell lesion of infectious bovine rhinotracheitis virus.TCID50 of NM strain determined by Reed-Muench method was 10-6.0/ml.NM strain could make neutralization test with IBRV positive serum,but it didn＇t react with IBRV negative serum.IBRV antibody-free calves with the age of 8 months showed typical clinical symptoms of IBR after inoculation of NM strain.The sequencing analysis results showed that the nucleotide homology between NM strain and K22 strain of IBRV.Conclusion] The research laid the foundation for developing the diagnosis reagent and its vaccine of IBR.