一株牛副流感病毒3型的分离鉴定及系统进化树分析

Isolation,identification and phylogenetic analysis of a bovine parainfluenza virus type 3 strain

目的分离鉴定一株疑似为牛副流感病毒3型(bovine parainfluenza virus type 3,BPIV3)的毒株,并分析系统进化树。方法从内蒙古某牛场患有呼吸道疾病的病牛鼻液中分离一株疑似为BPIV3的毒株,将分离毒株在MDBK细胞上进行增殖,并通过血凝试验及RT-PCR鉴定,同时对其N基因序列进行测定及分析,建立亲缘关系进化树。结果该分离病毒可在MDBK细胞上增殖,且可产生特异性细胞病变,第6代细胞的毒力为10^(7.1) TCID_(50)/100μL,与GenBank中已发表的BPIV3毒株N基因片段的同源性为99. 9%,与序列号为D84095. 1的BPIV3等亲缘性最近。结论本研究获得的分离株为BPIV3,与日本分离株(D84095. 1)同源性较高。

Objective To isolate,identify and perform phylogenetic analysis on a suspected bovine parainfluenza virus type 3(BPIV3)strain.Methods A suspected BPIV3 strain was isolated from the nose fluid of a cow with respiratory disease in a cattle farm in Inner Mongolia Mongolia Autonomous Region,China,propagated on MDBK cells,and identified by hemagglutination test and RT-PCR.The N gene of isolated BPIV3 strain was sequenced and analyzed,based on which a phylogenetic tree was constructed.Results The isolated BPIV3 strain was propagated on MDBK cells and caused specific CPE.The virulence of isolated strain of passage 6 was 10^7.1 TCID50/100 μL,while the homology was 99.9% to the N gene fragment of BPIV3 strain in GenBank,indicating the closest relation to the BPIV3 with sequence number of D84095.1.Conclusion The strain isolated in this study was BPIV3 with high homology to a strain(D84095.1)isolated in Japan.