多重巢式和定量PCR联合应用提高SARS病毒检测率

Multiplex Nested PCR Combined with Real-time PCR can Increase the Detecting Positive Rates of SARS-CoV

目的寻求一种可靠、可行的对急性严重呼吸道综合征病毒的早期诊断方法。方法应用多重巢式PCR联合荧光实时定量方法,对广州地区91份确诊和疑似病例标本、25份健康志愿者标本同时进行检测。结果25份健康志愿者标本经两种方法检测均为阴性。91份确诊和疑似病例标本经多重巢式PCR方法检测,阳性标本数53例,总阳性检测率58.24%;荧光实时定量PCR检测阳性标本数68例,阳性检出率74.73%;两种方法联合,阳性标本数76例,阳性检出率83.52%。结论多重巢式PCR联合荧光实时定量方法可进一步提高SARS病毒的阳性检出率,也为今后应对可能发生的其它突发性传染病病毒的检测提供分子诊断基础。

Objective To seek a reliable and feasible method to test severe acute respiratory syndrome virus as early as possible. Methods Test 91 samples of confirmed and/or possible cases and 25 samples of healthy volunteers simultaneously by combining multiplex nested PCR with real-time PCR. Results The detected samples of 25 volunteers were negative by these two methods; 53 out of 91 samples of SARS cases were positive by multiplex nested PCR and 68 out of 91 samples by real-time PCR, the positive rate was 58.24% and 74.73%, respectively. By combination of two methods, the positive samples were 76, the positive rate was 83.52%. Conclusion The results indicate these two methods have each advantage, complementary to each other. The positive rate increases by combining these two methods, which provide a rapid, sensitive, reliable approach to test SARS and provide the molecular diagnosis bases for detecting other possible sudden out break of virus diseases.