摘要
从河南省信阳市某猪场分离获得1株PRRSV,将病料处理接种Marc-145细胞,48 h后出现明显的细胞病变,通过RT-PCR和间接免疫荧光方法鉴定,证实分离到的毒株为PRRSV,暂命名为HN-XY12。进一步对其ORF5基因进行克隆和测序,并将其进行序列比对和遗传进化分析。结果表明:分离株ORF5基因与HP-PRRSV代表株JXA1、Hu N4和HEnan-1的核苷酸同源性分别为98.7%、98.8%和99.2%,且遗传进化距离很近,同处一个基因群中,而与CH-1a经典毒株较远,说明分离株属于HP-PRRSV毒株;进一步分析ORF5基因编码的GP5蛋白,第13、151位均为具有强毒特性的精氨酸(R),第137位为具有野毒特性的丝氨酸(S),且分离株与国内分离的高致病性毒株JXA1的GP5蛋白氨基酸序列基本一致,而与美洲型毒株VR2332相比有多处变异。该结论进一步说明了分离株为HP-PRRSV毒株。
Sample was collected from the farm in Xinyang of Henan province with suspected clinical signs of porcine reproductive and respiratory syndrome （PRRS）. Tissue sample that was crumbled and filtrated was inoculated into Marc-145 cells, PRRSV typical CPE was observed on MARC-145 cell, and then it was identified by RT-PCR and indirect immunofluorescenee assay （IFA）. The results showed that the stain was PRRSV, and was named PRRSV HE- XY12. The ORF5 gene of the isolate was amplified, cloned and sequenced for analyzing the genetic variation of PRRSV. Compared with JXA1, HuN4 and HEnan-1, the homologies of HN-XY12 were 98.7%, 98.8% and 99.2% in nucleotide, respectively; And the relatively distant relationship to HP-PRRSV strains and CH-1a strain was an independent branch. The results showed that PRRSV HN-XY12 strain belongs to HP-PRRSV. Further analysis GP5 of the sites of 13 and 151 were argrinine （R）whieh were related to virulent strains, and the site of 137 was serine（S） indicating that the isolated strain was wild strain; Compared with VR2332 strain, some variations were found in GP5 but the same with JXA1 strain. Furthermore the results showed that PRRSV HN-XY12 strain was HP-PRRSV.
出处
《中国畜牧杂志》
CAS
北大核心
2016年第19期71-75,共5页
Chinese Journal of Animal Science
基金
河南省科技攻关项目(162102110034)
关键词
PRRSV
分离鉴定
ORF5
变异分析
PRRSV
isolation and identification
ORFS
genetic variation analysis
