山羊源牛病毒性腹泻病毒的分离与鉴定

Isolation and identification of bovine viral diarrhea virus from goats

采集某山羊饲养场流产羔羊的组织样品,分离并鉴定牛病毒性腹泻病毒(BVDV)。经用BVDV5′-UTR引物进行PCR检测为阳性,将阳性样品处理后接种MDBK细胞,连续传代至第6代,RT-PCR检测每一代细胞中BVDV的P125基因,并进行序列测定。将第6代病毒液浓缩纯化,电镜观察病毒粒子。结果表明细胞培养物中含有BVDV,命名为JS12/02。将所得基因序列与其他国内外分离株的5′-UTR序列进行比对,系统进化分析表明该分离株属于BVDV-1型,这是国内首次报道从发生流产的山羊病料中分离到BVDV。

Tissue samples were collected from 6 stillborn fetuses with suspected bovine viral diarrhea （BVD） on a goat farm. RNA was extracted and examined with RT-PCR using two pairs of BVDV 5＇-UTR and BVDV P125 gene primers,respectively. Positive BVDV samples were inoculated in MDBK cells after treatment,and followed by 6 continuing passages,the cultures were tested by RT-PCR with the P125 gene primers and sequence analysis. The 6th cultures were concentrated, purified and examined with transmis- sion electron microscopy. The results indicated that the goats were positively infected by BVDV, named as JS12/02 strain. Phylogenetic analysis indicated that the isolated strain belonged to BVDV-1 through se- quence alignment with other strains of BVDV available in GenBank. One BVDV-1 virus strain was isolated for the first time from goats in an outbreak of abortion in China.