RT-PCR快速检测腮腺炎病毒

RT-PCR for rapid detection of mumps virus

目的 建立腮腺炎病毒的核酸快速诊断技术。方法 在腮腺炎病毒核蛋白基因 (NP)的保守区域设计一对半引物 ,采用逆转录聚合酶链 (RT -PCR)和半巢式PCR方法扩增腮腺炎病毒特异性核酸片段。结果 该方法能特异性地扩增腮腺炎病毒 3 5 7bp和 2 3 2bp的核酸片段 ,而对麻疹、风疹和流感病毒无交叉反应。检测腮腺炎病毒的灵敏度 ,1次PCR可检测出 10CCID50 ,2次PCR反应可达 0 1CCID50 。采用该方法从流行性腮腺炎患者含漱液标本中能直接检出腮腺炎病毒核酸条带。结论 该方法能特异、敏感地检测临床标本中腮腺炎病毒 。

Objective To develop the method of rapid detection of mumps virus RNA.Methods 1.5 pairs of primers in the nucleoprotein(NP) of the mumps virusg genome were designed.Reverse transcription polymerase chain reaction(RT-PCR) and semi-nested PCR were used to amplify specific fragments of mumps virus.Results 357?bp and 232?bp fragments of mump virus were amplified sensitively in the clinical specimens;however,no PCR products in measles,rubella and influenza viruses.The dilution experiments showed that mumps virus could be detected as few as 10CCID 50 in the first PCR and as few as 0.1CCID 50 in the second PCR.By using the method,mumps virus RNA were detected directly in the clinical specimens from patient with acute mumps infection.Conclusion RT-PCR was a rapid,sensitive and reliable method for the diagnosis of mumps infection.