摘要
从云南某猪场采取病料经RT-PCR检测为猪繁殖与呼吸综合征病毒(PRRS)阳性,将其处理后接种到猪肺泡巨噬细胞(PAM)和Marc-145细胞上。接种后3 d,猪肺巨噬细胞开始出现细胞病变(CPE);Marc-145细胞盲传3代后,细胞也出现了CPE。对出现CPE的细胞培养物,用RT-PCR法、间接免疫荧光试验进行检测,结果均为PRRSV阳性;细胞培养物经PEG初步浓缩后,接种到PRRSV阴性,PRV、CSFV抗体阳性的仔猪,15 d后猪只出现体温升高并伴有食欲减退,精神不振等症状。采血并用同上的RT-PCR法检测,结果为阳性;采血用同上的RT-PCR法检测,结果亦为阳性;用电镜对纯化后的病毒进行观察,可见有囊膜包裹的圆形病毒粒子,其直径约为50 nm;间接免疫荧光实验可见黄色荧光;TCID50测定其毒价为10-4.75/0.1 mL。该病毒对氯仿、紫外线、酸碱度变化(pH<5.0或pH>7.5)和热敏感;结果表明,已成功分离获得一株PRRSV云南地方流行毒株,命名为YN-1。
A strain of porcine reproductive and respiratory syndrome virus was isolated from infected domestic pigs from a Yunnan farm and shown PRRS positive in RT-PCR testing.The samples were filtered and then drained monolayers of PAM cells and Marc-145 cells,the typical cytopathic effect（CPE） was observed.15 days later,swines which had PRV,CSFV antibodies but lacked of PRRS antibodies showed typical syndrome of PRRS after inoculated with cell virus.RT-PCR technique was applied for the detection PRRSV in cells and showed PRRSV positive.The virus appeared typical porcine reproductive and respiratory syndrome virus form ：diameter was about 50 nm,circle with peplos and peplomer,by the indirect immunity fluorescence experiment,the yellow-grenn fluorescence occurred in the cell cytoblastema.TCID50 of virus was 10-4.75/0.1 mL-1.It was inactivated by chloroform,ultra-violet ray,pH5.0 or pH7.5 and heat.The result showed that PRRSV was indentified from Yunnan and it named YN-1.
出处
《动物医学进展》
CSCD
北大核心
2010年第4期11-15,共5页
Progress In Veterinary Medicine
基金
云南省自然科学基金项目(2006C0038M)
关键词
猪繁殖与呼吸综合征病毒
分离
鉴定
Porcine reproductive and respiratory syndromevirus virus
isolation
indentification
