山羊源伪狂犬病病毒的分离与鉴定

为确诊一例发病山羊是否感染伪狂犬病病毒,采集发病羊体的肺脏和脑组织进行伪狂犬病病毒gE基因PCR检测,并将病料接种至PK-15细胞分离病毒,以及进行小鼠感染试验和gD基因分析。结果表明,PCR检测结果 PRV阳性,病料接种PK-15细胞24h后,细胞开始出现细胞病变;将病毒感染小鼠,36h后小鼠出现局部奇痒、死亡;gD基因序列分析发现,分离毒株与GenBank中的PRV gD基因序列同源性均在98%以上,氨基酸同源性在99%以上;在分离毒株gD基因的808bp~837bp位置上存在缺失与变异、高变重复区。本研究成功分离获得一株羊源伪狂犬病毒,为云南省羊伪狂犬病防控和基础研究提供资料。

Isolation and Identification of PRV from Goat

To detect whether a goat was infected with pseudorabies,the lung and brain of the diseased sheep were collected to carry out the PCR detection of gE gene of the pseudorabies virus;and the material was inoculated into PK-15 cells to isolate the virus;as the same time,the mice infection test and gD gene analy-sis were made.The result of PCR showed that PRV was positive.After 24 hours of inoculation,the cells began to show cytopathic effect;the mice infected with virus showed partial itching and death after 36 hours.Furthermore,gD gene sequence analysis showed that the homology of PRV gD gene in GenBank was more than 98% and the homology of amino acid was more than 99%.The deletion and mutation of gD gene in 808 bp-837 bp locus were found in the isolates.In this study,pseudorabies virus was successfully isola-ted from the goat,which provided data and basic research for prevention and control of goat pseudorabies virus in Yunnan province.