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目的选择合适的RTPCR检测方法应用于人和与人类生活密切相关的多种野生、圈养和市售动物携带SARS样冠状病毒的检测,探索SARS样病毒的来源。方法收集深圳SARS临床诊断病例、疑似及观察病例咽漱液标本共350份,分别用TaqMan和分子信标荧光RTPCR法进行检测;收集386只动物的咽肛拭子样本及病毒分离培养物共442份样品进行了TaqMan荧光RTPCR和普通RTPCR法的检测。结果41例临床确诊病例荧光RTPCR法检出10份阳性,阳性率24.39%,TaqMan法和分子信标法检出结果8份符合(符合率88.89%)。对动物样本病毒分离培养物细胞病变样本18份进行检测,检出16份阳性,其中14份(87.5%)样本来源为果子狸。深圳东门市场及广东周边地区市场及养殖场动物样本检测结果:果子狸、貉及獾类动物总的阳性率为39.02%,而其它动物的阳性率为0,两者间差异具有显著性(P<0.01);对东门市场109只主要野生动物咽肛拭子样本进行PCR检测,阳性率44.04%,而145只野外的野生动物阳性率仅为0(P<0.01)。结论TaqMan法和分子信标法均可用于SARS检测;动物中尤其是果子狸等野生动物可能携带有SARS冠状病毒。
Objective To select suitable RTPCR detection methods to detect human SARS-like coronavirus (SARS-like coronavirus) and detect the origin of SARS-like viruses in humans and other wild, captive and commercial animals closely related to human life. Methods A total of 350 samples of SARS cases were collected from suspected and observed SARS patients in Shenzhen. The TaqMan and molecular beacon fluorescence RTPCR methods were used to detect the samples. A total of 442 samples of pharyngeal swabs from 386 animals were collected. Samples were tested by TaqMan fluorescent RTPCR and conventional RTPCR methods. Results Of the 41 clinically diagnosed cases, 10 were positive by fluorescence RTPCR and the positive rate was 24.39%. The results of TaqMan assay and molecular beacon assay were 8 (88.89%). Twenty-eight samples of cytopathic effect were isolated from animal samples, and 16 samples were positive. Among them, 14 samples (87.5%) were from civens. Shenzhen Dongmen Market and Guangdong surrounding market and farm animal sample test results: the total positive rate of civet cats, catfish and badger animals was 39.02%, while the positive rate of other animals was 0, the difference was significant P <0.01). PCR was carried out on pharyngeal and anal swab samples from 109 main wild animals in Dongmen market. The positive rate was 44.04%, while the positive rate of wild animals in 145 fields was only 0 (P <0.01). Conclusion Both TaqMan method and molecular beacon method can be used for SARS detection. Animals such as civets may carry SARS coronavirus.