荧光PCR、常规PCR和细胞培养在流感检测中的对比分析

Comparative analysis of fluorescent PCR,conventional PCR and cell culture in detection of influenza virus

目的比较当地已有的3种流感病毒检测方法的差异,以确定它们在不同情形下的适用方向。方法通过荧光PCR(聚合酶链反应)、常规PCR和细胞培养3种方法同时检测413份流感样病例(ILI)咽拭子样本,计算出它们的灵敏度和特异度,并进行统计分析。结果荧光PCR、常规PCR和细胞培养的阳性检测率分别为20.34%、15.25%、11.86%;相对于荧光PCR,常规PCR的灵敏度和特异度分别为75%、100%,细胞培养的灵敏度和特异度分别为56%、99.39%;与常规PCR相比,细胞培养的灵敏度为75%,特异度为99.43%。结论 3种方法在灵敏度方面相差很大,但在特异性上几乎没有差异。由于荧光PCR和常规PCR具有很高的灵敏度,并且检测速度快,适用于暴发疫情的应急诊断;细胞培养作为一种经典的病原学检测方法,可以获得流感毒株,用来开展抗原性、基因特性和耐药性等深入研究。

Objective To compare the differences among three influenza virus detection assays being used in local laboratory, in order to determine their appropriate application fields. Methods Four hundred and thirteen throat swabs samples from influ- enza- like illness（ILI） eases were detected by fluorescent PCR（ Polymerase Chain Reaction） , conventional PCR and cell cul- ture simultaneously, then the sensitivity and specificity of these methods were calculated and assessed by statisties. Results The positive ratios of fluorescent PCR, conventional PCR and cell culture were 20.34% , 15.25% , 11.86% respectively. In contrast to fluorescent PCR, the sensitivity and specificity of conventional PCR were 75% and 100% respectively, while those of cell culture were 56% and 99.39%. Cell culture showed sensitivity and specificity of 75% and 99.43% compared with con- ventional PCR. Conclusion There was significant difference in sensitivity but almost no in specificity among the three meth- ods. With high sensitivity and fast testing velocity, fluorescent PCR and conventional PCR are suitable for rapid diagnosis in ep- idemie outbreak. Cell euhure, as a classical method for etiologieal surveillanee, can isolate influenza virus and conduct inten- sive studies on antigenicity, genetic characteristies and drug resistance.