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[目的]建立一种特异、灵敏、快速检测脑膜炎奈瑟菌的实时荧光PCR方法。[方法]针对脑膜炎奈瑟菌特异性基因porA设计引物、探针,在实时荧光PCR仪上进行扩增、检测和结果分析;用建立的检测方法与常规PCR方法的检测结果做对比。[结果]应用该实时荧光PCR方法,64株脑膜炎奈瑟菌检测结果均为阳性,6株非脑膜炎奈瑟菌结果均为阴性;检测灵敏度达5.0×l03cfu/ml,即5cfu/test,比常规PCR方法灵敏度高10倍;该方法重复性好。[结论]实时荧光PCR方法检测脑膜炎奈瑟菌具有灵敏、特异、快速方便的特点,可用于定量检测。
[Objective] To establish a specific, sensitive and rapid real-time fluorescence PCR method for the detection of Neisseria meningitidis. [Methods] Primers and probes were designed for Neisseria meningitidis-specific gene porA. The amplification, detection and result analysis were carried out on the real-time fluorescence PCR instrument. The established detection method was compared with the conventional PCR method. [Results] The results of 64 real-time PCR assays showed that Neisseria meningitides were positive in all 64 strains of Neisseria meningitidis and negative in 6 strains of Neisseria meningitidis. The detection sensitivity was 5.0 × 10 3 cfu / ml (5 cfu / test) Sensitivity than conventional PCR method 10 times higher; the method is good repeatability. [Conclusion] The real-time fluorescent PCR detection of Neisseria meningitidis is sensitive, specific, rapid and convenient, and can be used for quantitative detection.