应用多重荧光定量RT-PCR技术检测麻疹病毒的适用性分析

The applicability analysis of detecting measles virus by multiplex real-time fluorescent PCR

目的验证多重荧光定量RT-PCR方法用于麻疹病原学诊断的准确性,探讨其在麻疹快速检测方面的适用性。方法对吉林省2013年的149例麻疹疑似病例咽拭子标本,进行多重荧光定量RT-PCR、双重荧光定量RT-PCR实验,对52份获得麻疹病毒序列的标本进行多重荧光定量RT-PCR和序列分析实验,并将2组实验结果分别进行对比分析。结果多重荧光定量RT-PCR检测结果与麻疹/风疹双重荧光定量RT-PCR检测结果一致,均得到140份麻疹阳性标本、9份阴性标本,一致率为100%;多重荧光定量RT-PCR检测结果与序列分析结果均显示JL/M-13-3号标本为疫苗株A基因型,其他51份标本为野毒株H基因型,结果吻合率为100%。结论多重荧光定量RT-PCR是一种快速、灵敏、准确的实验方法,适用于快速诊断麻疹病例、及时鉴别麻疹H基因型野病毒和A基因型疫苗株引起的病例。

Objective To verify the etiological diagnostic accuracy of measles virus by multiplex Real - time fluorescent PCR, so as to explore its applicability to the quick detection of measles. Methods All the 149 throat swabs of the suspecious patients from Jilin province in 2013 were tested respectively by multiplex real -time fluorescent PCR and dual - channel real - time fluorescent PCR. The 52 throat swabs receiving their sequences were tested by muhiplex real - time fluorescent PCR and sequencing analysis, and then, the test results were compared. Results The outcome of n）uhiplex real - time fluorescent PCR were consistent with dual - channel＇ s. 140 cases were positive and 9 cases were negative, with the coincidence rate of 100%. Both multiplex Real-time fluorescent PCR and sequence assays showed that NO. JL/M -13 -3 was measles vaccine virus genotype A, and the other 51 were measles wild virus genotype H, with the consistency rate up to 100%. Conclusion Multiplex real - time fluorescent PCR method is rapid, sensitive and accurate. This method is applicable to rapid diagnosis, and timely identification of the measles virus cases caused by wild virus H genotype and vaccine virus A genotype.