麻疹病毒、风疹病毒和腮腺炎病毒多重RT-PCR法的建立

Multiplex RT-PCR for simultaneous detection of masern,rubella and mumps viruses

目的建立针对麻疹病毒、风疹病毒和腮腺炎病毒的多重RT-PCR检测方法。方法分别根据麻疹病毒M基因、风疹病毒E基因和腮腺炎病毒M基因设计3对引物,建立同时检测3种病毒的多重RT-PCR,并评估其灵敏度和特异性。结果建立的多重RT-PCR可同时或分别扩增的3种病毒的111 bp、352 bp和274 bp基因片段,其灵敏度分别达到2.1、2.1、2.2lgCCID 50/mL;而与脊髓灰质炎病毒和乙型脑炎病毒无交叉反应。结论建立的多重RT-PCR可用于麻疹病毒、风疹病毒和腮腺炎病毒的快速检测。

Objective To establish a multiplex RT-PCR method for simultaneous detection of masern, rubella and mumps viruses. Methods Based on the sequences of masern virus M gene, rubella virus E gene and mumps virus M gene, three sets of specific primers were designed and the multiplex RT-PCR for simultaneous detection of the 3 viruses were established. The specificity and sensitivity of the developed multiplex RT-PCR method were evaluated. Results Three specific fragments, including masem virus M gene （ 111 bp）, rubella virus E gene （352 bp） and mumps virus M gene （274 bp） , were amplified successfully by the multiplex RT-PCR method. The sensitivities were 2.1, 2.1 and 2.2 lg_CCID 50/mL respectively. No cross reaction with poliovirus or Japanese encephalitis virus was found. Conclusion The established muhiplex RT-PCR could rapidly and simultaneously detect the infection of masern, rubella and mumps viruses.