逆转录套式PCR检测粪便样本中的SARS冠状病毒

Detection of SARS-CoV from feces samples with RT-PCR

为了观察SARS冠状病毒在SARS患者粪便中的存在规律,建立了检测SARS冠状病毒RNA的逆转录聚合酶链反应(RTPCR)方法,并应用该方法检测了241份SARS患者粪便样本。部分PCR产物应用测序技术进行验证。RTPCR的灵敏度为1010稀释度的病毒原液(原液为108TCID50/ml)。241份粪便样本的总体检出率为24.1%(58/241),其中发病后的前10d和20d的检出率均为50.0%。随着发病时间的延长,阳性检出率呈下降趋势。应用RTPCR从粪便中检测SARS冠状病毒是可行的,在发病50d以后仍有17.0%左右的阳性检出率,提示SARS恢复期患者具有排毒的可能性,给后续的卫生防疫措施提供了一定的参考数据。

In order to study the existing status of SARS CoV in feces samples, reverse transcript nested PCR (RT PCR) was established and 241 feces samples were detected with this method. The sensitivity for detection of SARS CoV(10 8 TCID 50 /ml) in supernatant of culture from infected Vero E6 cells was 10 -10 dilution. Part of the PCR products were sequenced and confirmed to be the specific sequences of SARS CoV. The overall detection rate of 241 feces samples was 24.1 %( 58 /241). For samples from 0 to 20 days after onset of symptom, the detection rate of SARS CoV RNA was 50.0% . The detection rate of SARS CoV RNA in feces samples became lower as the days after onset of symptom became longer, but SARS CoV RNA was still detected in 16.7% feces samples from patients on more than 50 days after onset of symptom. This provide a reference information for public health prophylaxis.