猪繁殖与呼吸综合征病毒新疆株的分离与鉴定

Isolation and Identification of Porcine Reproductive and Respiratory Syndrome Virus of Xinjiang

为了解猪繁殖与呼吸道综合征在新疆的流行现状,以及病毒毒株的分子生物学特征,对疑似该病病料进行地方毒株的分离,并对其进行鉴定。采集新疆乌鲁木齐市七道湾某猪场疑似PRRS病死的猪只肺脏及淋巴结等组织病料,将其处理后接种到Marc-145细胞上,并盲传3代;对分离株进行毒力测定(TCID50),应用RT-PCR方法对出现CPE的细胞培养物进行分子检测。结果显示,分离到的新疆病毒株可发生细胞病变,在Marc-145细胞上的TCID50为10-5·mL-1。RT-PCR检测结果用琼脂糖凝胶电泳鉴定基因序列,将测序结果与GenBank已发表的PRRSV毒株基因序列进行对比分析。研究证明所分离到的病毒为PRRSV,命名为XJ-Q。

In order to understand the prevalence status and molecular biology characteristics of strains of porcine reproductive and respiratory syndrome（PRRS） in Xinjiang, an experiment was carried out to isolate reproductive and respiratory syndrome virus（PRRSV） from the specimen of suspicious cases. The suspicious lungs and lymphoid tissue samples collected from dead piglets on a pig farm in Qidaowan of Urumqi, Xinjiang. Firstly, the simples were processed in the laboratory, and then they were incubation to Marc-145 cells. After this procedure, these cells reproduced blindly for three generations; toxicity（TCID50） serum neutralization test of isolated strains were conducted, by RT-PCR method for the occurrence of cell cultures for molecular detection of CPE. The results showed that Xinjiang strains of it could cause cells pathological changes with the 10^-5·mL^-1TCID50. RT-PCR reaction result by agarose gel electrophoresis to identify gene sequences. The result of sequencing and analyzing with the PRRSV searched in GeneBank.It was confirmed that the eight isolated strains were PRRSV, named XJ-Q.