猪痘病毒江西分离株JX08的分离鉴定  

Isolation and identification of swinepox virus Jiangxi isolate JX08

作　　者：邓舜洲[1] 徐昌满[1] 张文波[1] 蒋新华[1] 邹柳[1] 冷闯[1] 胡杨[1] 朱芝秀[1] 

机构地区：[1]江西农业大学动物科学技术学院,江西南昌330045

出　　处：《中国兽医学报》2014年第5期700-703,共4页Chinese Journal of Veterinary Science

基　　金：江西省科技支撑计划项目(20132BBF60044);江西省教育厅项目(GJJ13287)

摘　　要：采用PK15细胞培养,对江西省某猪场疑似猪痘的病猪皮肤丘疹进行病毒分离,分离到1株猪痘病毒,命名为SWPV JX08,。根据猪痘病毒基因组序列设计了5对引物,对分离毒株进行TK基因、P35基因、P23基因、TK基因上游和下游片段分别进行PCR扩增,扩增片段与GenBank收录的猪痘病毒（AF410153.1）的核苷酸序列的同源性分别为97.5%、98%、98.6%、98.2%和98%。该分离毒株不但能使PK15细胞产生稳定的细胞病变,还能适应非猪源性的Vero细胞,并产生细胞病变;电镜下可见感染的PK15细胞内形成椭圆形或圆形的包涵体,胞浆中有大量椭圆形、砖形的典型猪痘病毒粒子,病毒粒子中央为哑铃状芯髓,病毒粒子大小为（244~340）nm×（164~263）nm。该毒株皮下或静脉接种25日龄仔猪,感染7~11d后,表现典型猪痘的症状（皮肤出现丘疹）,其血清中可检测到SWPV抗体。A field strain of swinepox virus （SWPV）, designated as SWPV J X08 strain, was isolated by PK15 cells cultures from skin pustules and vesicles on nursery pigs in Jiangxi province. The homology of TK,P35,P23 genes,and TK gene upstream and downstream region nucleotide sequence between isolated virus JX08 and SWPV reference strain （AF410153.1） were analyzed by DNAStar,indicating that the homologies of nucleotide of the above gene in both SWPV P35 strains was 97.5% ,98 ~//0,98.60//00,98.2% and 98% ,respectively. The JX08 strain was successfully adapted to the PK15 and Vero cell lines with typical viral cytopathic effects （CPE）. A large num- ber of mature SWPV particles were observed in the cytoplasm infected with JX08 strain,exhibiting a ellipse or brick-like structure approximately （244-340） nmX （164-263） nm. The virion was composed of a central biconcave core by two ellipsoid lateral bodies. The infected 25-days old pig- lets showed typical swine pox skin lesions, such as papulae, pustules and vesicles,and the P35 protein antibodies of SWPV were detected by ELISA method in the sera.

关 键 词：猪痘病毒 分离鉴定 江西分离株JX08 

分 类 号：S852.65[农业科学—基础兽医学]