牛副流感病毒3型的分离鉴定

Isolation and Identification of Bovine Parainfluenza Virus Type3

摘要:

从黑龙江省某牛场患有呼吸道疾病的病牛鼻液中分离到1株病毒, 该病毒能在MDBK细胞上增殖并产生特征性细胞病变, 对分离毒株进行理化特性、RT-PCR鉴定及序列测定与分析, 结果发现该病毒为RNA病毒, 具有血凝性, 对乙醚、氯仿极其敏感, 不耐热, 对酸的抵抗力差; 分离株与GenBank中已发表的病毒株N基因片段的同源性为82.6%~99.1%, 结果表明分离的病毒为牛副流感病毒3型毒株, 命名为BPIV3 DQ1株。根据GenBank上发表的牛副流感病毒3型核衣壳蛋白N基因序列, 设计合成了一对特异性引物, 能扩增704 bp的目的片段, 可以检测出10个 TCID50/100 μL病毒核酸。

Abstract:

One virus strain was isolated from the nasal secretion of a bovine suspected to be respiratory tract disease at a beef farm in Heilongjiang province. The virus could grow well and produce typical cytopathic effect in MDBK cells. Bovine parainfluenza virus type3(BPIV3) has been identified by a series of systematic identification such as physicochemical features, RT-PCR, and sequence analysis. As a result, the virus is an RNA virus, which is of hemagglutination and is very sensitive to ether and chloroform, no resistance to heat and acid. The homology of the nucleocapsid(N) gene sequence of the isolate and of GenBank is 82.6% to 99.1%. The isolated virus was confirmed as Bovine parainfluenza virus type3 named BPIV3 DQ1 strain. Specific primers were designed and synthesized according to the reported N protein gene sequence of Bovine parainfluenza virus type3 in GenBank. The size of 704 bp fragment was amplified and 10 TCID50/100 μL of viral nucleic acid was detected.