猪繁殖与呼吸综合征病毒传统毒株与变异毒株的鉴别诊断  

Differentiated Detection of Conventional from Variant Isolates on Porcine Reproductive and Respiratory Syndrome Virus

作　　者：于美芳[1,2] 吴家强[2] 张金强[2] 李俊[2] 张秀美[2] 景田军 王雪红[4] 王金宝[1,2] 

机构地区：[1]青岛农业大学动物科技学院,山东青岛266109 [2]山东省农业科学院畜牧兽医研究所,山东济南250100 [3]山东牧工商总公司,山东济南250100 [4]山东出入境检验检疫局,山东青岛266001

出　　处：《家畜生态学报》2009年第2期84-87,共4页Journal of Domestic Animal Ecology

基　　金：山东省自然科学基金重点项目(Z2007D06);山东省重大科技专项(2007ZHZX11103);山东省农科院青年基金项目(2005YQ039)

摘　　要：从山东各地猪场采集了50份疑似猪繁殖与呼吸综合征(PRRS)感染的病料,采用RT-PCR技术扩增出PRRS病毒(PRRSV)的ORF7基因和Nsp2基因,并对Nsp2基因进行了序列同源性比较和遗传进化树分析,从RT-PCR阳性病料中进行病毒分离,然后应用Nsp2单克隆抗体对分离毒株进行鉴别诊断。结果表明,从8份病料中同时扩增出病毒的ORF7基因和Nsp2基因,序列分析显示其中6株病毒的Nsp2缺失30个氨基酸,与Nsp2单克隆抗体不发生反应,属于变异毒株;另外两株病毒的Nsp2未见氨基酸缺失,与Nsp2单克隆抗体发生特异性反应,属于传统毒株。序列同源性和系统进化分析表明,6株变异毒株与其他高致病性毒株亲缘关系很近,而与经典毒株的遗传距离较远。A total of 50 samples which were suspected with porcine reproductive and respiratory syndrome were collected （PRRS） in Shandong area. The ORF7 gene and Nsp2 gene of PRRS virus （PRRSV） were amplified by RT--PCR, and nucleotide homolog and phylogenetic analysis were performed based on the Nsp2 gene of PRRSV. Viruses were isolated from PCR--positive samples and differentiation between conventional and variant PRRSV was conducted using monoclonal antibody （MAb） against Nsp2. It showed that ORF7 gene and Nsp2 gene were amplified from 8 samples, and 6 isolates with 30 amino acids deletion in Nsp2 were identified as variant PRRSVs and had no reactivity with MAb against Nsp2. The other 2 isolates with no deletion in Nsp2 were identified as conventional PRRSVs and had strong specific reactivity with Nsp2. The results of nucleotide homology and phylogenetic analysis showed that the six variant PRRSV isolates were relatively close to the other highly pathogenic viruses in comparison with the conventional isolates.

关 键 词：猪 繁殖与呼吸综合征病毒 变异株 传统株 鉴别诊断 

分 类 号：S811.5[农业科学—畜牧学]