猪繁殖与呼吸综合征病毒JX0708株的分离鉴定及其Nsp2基因的遗传变异分析  

Isolation and Identification of PRRSV JX0708 Strain,and Analysis on Genetic Variation of Nsp2 Gene

作　　者：贾怀杰[1] 陈国华[1] 房永祥[1] 王晓霞[1] 李小庆[1] 莫斯科[1] 瞿惠玲[1,2,3] 景志忠[1] 

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部兽医公共卫生重点实验室,兰州730046 [2]甘肃农业大学动物医学院,兰州730070 [3]甘肃省兽医局,兰州730030

出　　处：《中国畜牧兽医》2010年第10期58-63,共6页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然基金项目(30871884);国家支撑计划(2006BAD31B03);甘肃省支撑计划(0804NKCA076)

摘　　要：采集江西省某猪场疑似猪繁殖与呼吸综合征病料,RT-PCR检测可扩增出PRRSV ORF7特异性片段,序列分析发现其与美洲型毒株序列一致性达93.9%以上,与欧洲型毒株仅为69.8%。将采集的脑和肺脏处理后接种Marc-145细胞盲传,在盲传的细胞中可扩增到PRRSV ORF7特异性片段,传至第6代时出现明显的细胞病变;将该毒株NSP2非结构蛋白基因测序分析,发现其编码的氨基酸与国内近年来分离的高致病性PRRSV毒株有较高的同源性,与美洲型代表株VR-2332相比缺失第482位和533-561位共30个氨基酸;纯化病毒并进行电镜观察,可见典型的PRRSV病毒粒子。第8代的细胞毒回归30日龄仔猪,可观察到猪繁殖与呼吸综合征典型症状和病理变化。这表明已成功地分离到1株美洲型PRRSV变异株,命名为JX0708株。Suspected specimens collected from pigs with porcine reproductive and respiratory syndrome virus（PRRSV） in Jiangxi province,was ORF7 gene positive tested by RT-PCR.The result of sequence analysis showed that ORF7 gene of this strain shared 93.9%,69.8% homologies with those of American standard strain and European strain.Brains and lungs from clinical specimens had regular cytopathic effect（CPE） over 6 passages on Marc-145 cell culture,ORF7 gene can be detected at cell cultures;sequence analysis of Nsp2（non-structural protein） gene showed that there was a high amino acid homology between JX0708 and other Chinese local PRRSV strains isolated in recent years,while this strain had 30 amino acid deletion at the residues 482 and 533 to 561 compare with the American standard strain.Electron microscopy demonstrated that typical virions were found in the purified cell cultures.Cell cultures of 8th passages was used to do regression test on the piglets of 30 day-old.After the inoculation,all piglets showed typical clinical signs and patiological changes of PRRS.So an America variant PRRSV stain was isolated successfully,and named JX0708 strain.

关 键 词：猪繁殖与呼吸综合征病毒 JX0708株 分离鉴定 NSP2基因 遗传变异分析 

分 类 号：Q78[生物学—分子生物学]