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目的建立TaqMan-MGB探针荧光定量PCR快速检测脑膜炎奈瑟菌(Nm)方法,为流脑的诊断提供更加灵敏、特异、可标准化、自动化的检测方法;评价TaqMan-MGB探针荧光定量PCR法在健康人群带菌调查中的应用价值。方法根据Nm ctrA基因的3端保守序列设计合成引物和TaqMan-MGB探针,建立快速检测脑膜炎奈瑟菌的荧光定量PCR方法,并对反应条件进行优化。结果TaqMan-MGB探针荧光定量PCR检测脑膜炎奈瑟菌的灵敏度为102cfu/ml,TaqMan-MGB探针法检测900份健康人群咽拭子标本,阳性34例,检出率3.8%,高于培养法(1.2%)。结论该方法特异性强、灵敏度高,能实现脑膜炎奈瑟菌的快速检测,适用于健康带菌检查并可作为临床常规诊断方法的补充,有利于流脑的早诊断、早治疗。
Objective To establish a TaqMan-MGB probe for rapid detection of Neisseria meningitides (Nm) by fluorescence quantitative PCR and to provide a more sensitive, specific, standardized and automated detection method for the diagnosis of meningoencephala. To evaluate TaqMan-MGB probe quantitative PCR Application Value of Law in the Survey of Healthy Population. Methods The primers and TaqMan-MGB probes were designed according to the 3-terminal conserved sequence of Nm ctrA gene. A rapid quantitative PCR method was established for the detection of Neisseria meningitides. The reaction conditions were optimized. Results The sensitivity of TaqMan-MGB probe for detection of Neisseria meningitides was 102cfu / ml, and that of throat swabs in 900 healthy people by TaqMan-MGB probe was 34, with a positive rate of 3.8% Training method (1.2%). Conclusion The method is specific and sensitive and can detect Neisseria meningitidis rapidly. It is suitable for healthy infection test and can be used as a supplement to routine clinical diagnosis. It is good for early diagnosis and early treatment of.