论文部分内容阅读
目的:从粪便标本中分离并鉴定SARS-CoV. 方法:粪便浸出液接种Vero-E6细胞,盲传至出现明显的细胞病变效应(CPE),并能稳定传代.采用电镜法和RT- PCR技术鉴定细胞培养物中的SARS-CoV(本实验均在BSL-3实验室完成). 结果:大部分样本用Vero-E6细胞盲传3代后发现明显的CPE,并能稳定传代.电镜负染检查,出现CPE的细胞可查见病毒颗粒;培养物上清RT-PCR扩增均为阳性.粪便样本直接用RT-PCR检测,阳性率仅为50%,细胞培养的检出率为73%. 结论:采用细胞培养结合RT-PCR技术检测粪便标本SARS- CoV比粪便标本直接RT-PCR的阳性率高.
OBJECTIVE: To isolate and identify SARS-CoV from stool specimens.Methods: Vero-E6 cells were inoculated into exudates of stool samples, and the cells were stably passaged after blind passage to CPE.Electron microscopy and RT-PCR SARS-CoV in cell culture (all of the experiments were performed in BSL-3 laboratory) .Results: Most of the samples found significant CPE after three generations of blind passage in Vero-E6 cells and could stably pass the test. , CPE cells could be seen virus particles; culture supernatants were positive by RT-PCR amplification.The stool samples were directly detected by RT-PCR, the positive rate was only 50%, the detection rate of cell culture was 73%. Conclusion: The positive rate of SARS-CoV in stool samples detected by cell culture combined with RT-PCR was higher than that of stool specimens by direct RT-PCR.