实时荧光RT-PCR法快速检测麻疹病毒核酸

Rapid detection of measles virus RNA by real-time fluorescent RT-PCR assay

目的采用实时荧光RT-PCR法快速检测麻疹病毒核酸,早期确诊麻疹疑似病例,为及时采取防控措施,减少麻疹的暴发流行提供科学的实验依据。方法对沈阳市2013年4月-11月63份麻疹疑似病例,在出疹3 d内采集静脉血和咽拭子。血清标本采用ELISA法检测麻疹Ig M抗体,咽拭子标本采用实时荧光RT-PCR法检测麻疹病毒核酸,核酸检测阳性的标本进行病毒分离。结果 63份麻疹疑似病例中麻疹病毒Ig M抗体有27份为阳性,阳性率为42.86%;麻疹病毒核酸有32份为阳性,阳性率为50.79%;核酸检测阳性标本中,4份分离到麻疹病毒,分离率为12.50%。麻疹Ig M抗体检测和实时荧光RT-PCR检测同为阳性27份,同为阴性31份;实时荧光RT-PCR检测阳性而麻疹Ig M抗体检测阴性的为5份,2种方法检测的总符合率为92.06%。结论实时荧光RT-PCR法是一种灵敏、准确、安全的快速检测麻疹病毒核酸的方法,可用于麻疹病毒感染的早期诊断。

Objective Early diagnosis of suspected measles cases through rapid detecting measles viral RNA by real - time flu- orescence RT - PCR assay to provide scientific experimental basis for taking immediate measures to control outbreak of measles. Methods Venous blood and throat swab specimens were collected from 63 suspected measles cases in Shenyang from April to November in 2013. All specimens were collected within the 3 days after rashes occurred. Serum measles IgM was detected by ELISA and measles viral RNA from throat swab specimens was detected by real - time fluorescence RT - PCR assay. Measles viral was isolated from the throat swab specimens with positive results in viral RNA detection. Results 63 cases of suspected measles, 27 measles virus IgM antibody detected by ELISA method was positive with the positive rate of 42.86% ; 32 measles virus nucleic acid was positive with the positive rate of 50.79%. There were 4 positive specimens in measles virus detection from measles viral RNA positive specimens and the isolation rate of measles virus was 12.50%. Positive specimens from both measles IgM and viral RNA were 27 and negative specimens from both measles IgM and viral RNA were 31. Real time fluores- cent RT - PCR positive detection, and measles IgM antibody negative detection of 5 copies, the total compliance rate of the two methods was 92.06%. Conclusion Real - time fluorescence RT - PCR can be used as a early diagnosis method for measles due to its sensitiveness, correctness and safety.