实时荧光定量PCR技术用于脑膜炎奈瑟菌的快速检测

Real-time fluorescent quantitative PCR for detection of Neisseria meningitidis

目的:建立流行性脑脊髓膜炎的实时荧光定量PCR法,考察其在流脑检测中的应用。方法:采用国家标准的细菌培养法对167份流脑监测标本进行病原体筛查,同时采用实时荧光定量PCR方法对该167例标本进行检测,比较两者的实验结果,并考察实时荧光定量PCR方法的特异性和灵敏度。结果:实时荧光定量PCR检测结果与传统检测方法的阳性符合率为55.56%,除完全符合的5例标本之外,另有4例采用传统方法检测为阴性的标本用实时荧光定量PCR检测显示为阳性,表明实时荧光定量PCR方法具有更高的灵敏度;特异性和灵敏度实验表明实时荧光定量PCR方法的特异性达100%,灵敏度为103拷贝/ml。结论:实时荧光定量PCR法具有准确性好、灵敏度高、检测快速等优点,是一种理想的脑膜炎奈瑟菌检测方法。

Objective：To establish a method of fluorescent quantitative PCR and examine its utility for detection of Neisseria meningitidis.Methods：167specimens were detected by culture method of national standards,and compared with the method of fluorescent quantitative PCR.The results of two different methods were compared and the sensitivity and specificity of fluorescent quantitative PCR method was obtained.Results：The detective result of fluorescent quantitative PCR was 55.56% accordance to that of traditional methods.5 specimens were detected positive both by traditional methods and fluorescent quantitative PCR method,and 4 specimens detected negative by traditional methods were positive by fluorescent quantitative PCR method,which showed that fluorescent quantitative PCR method had higher sensitivity than traditional methods.The specificity and sensitivity of fluorescent quantitative PCR method was 100% and 103 copies/ml,respectively.Conclusion：Real-time fluorescent quantitative PCR is an ideal method for detection of Neisseria meningitidis with high accuracy and sensitivity.