用RT-PCR和实时荧光RT-PCR检测诺沃克病毒的比较分析

Comparative analysis of methods of RT-PCR and real-time RT-PCR for detection of Norwalk-like virus

目的:研究用不同方法检测诺沃克病毒核酸的阳性率关系,为进一步完善诺沃克病毒核酸的实验室检测提供依据。方法:采用逆转录聚合酶链反应(RT-PCR)及实时荧光逆转录聚合酶链反应(real-tim e RT-PCR)两种方法,对124例腹泻标本进行诺沃克病毒核酸的检测。结果:124份标本中,RT-PCR检出诺沃克病毒阳性31例,阳性率为25%;实时RT-PCR检出阳性37例,阳性率为29.84%;两种方法均阳性的28例,均阴性的84例,符合率为90%;不同性别阳性率无显著性差异;采用实时RT-PCR检测诺沃克病毒,敏感性较RT-PCR高,但两种方法的阳性率无统计学差异。对于强阳性样本(Ct值在20以内或电泳结果特异性条带很亮),两种方法检出率均为100%;对于弱阳性样本,则大部份标本实时RT-PCR检出阳性而RT-PCR未能检出,少部份标本RT-PCR检出阳性而荧光RT-PCR未能检出。结论:对于诺沃克病毒的实验室检测方法,首选实时RT-PCR;在可能的情况下,应该RT-PCR和实时RT-PCR两种方法联用进行检测,以避免发生弱阳性样本漏诊的情况。

Objective：To improve the detective method of Norwalk-like virus and establish the database about the detection in laboratory, we researched the positive rate of Norwalk - like virus RNA with different methods. Methods ： Detected 124 diarrhea samples with RT- PCR and the real-time fluorescent quantitative RT-PCR. Results： The positive detection rate of RT-PCR of 124 diarrhea samples was 25%, and the positive detection rate of real- time RT-PCR was 29. 84%. The identical positive samples were 28, and the identical negative samples were 84 of the 124 samples, the coincidence rate was 90%. There were no statis-tical difference between male and female. Compared with RT-PCR, the sensitivity of real-time RT-PCR was higher, but the positive rate of two methods did not have marked difference. The detective rates of strongly positive samples of the two methods were 100%, and the detective rate of weakly positive samples with real-time RT-PCR method was higher than the result of RT-PCR. Conclusion：The first choice of Norwalk-like virus detection method should be the real-time RT-PCR and it is better to detect the samples with RT-PCR and real-time RT-PCR in order to diagnose the weakly positive samples.