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目的建立8种细菌性脑膜炎多重聚合酶链反应(Multiplex-polymerase Chain Reaction M-PCR)检测方法。方法针对脑膜炎奈瑟菌、流行性感冒嗜血杆菌、肺炎链球菌、金黄色葡萄球菌、单核细胞增生性李斯特菌、猪链球菌、结核杆菌、新生隐球菌的种特异性基因,通过优化单一PCR扩增体系及扩增程序,建立M-PCR方法检测8种导致细菌性脑膜炎的病原体,检测M-PCR体系的灵敏度。结果初步建立了检测8种导致细菌性脑膜炎病原体的M-PCR扩增方法,能够同时检测多种导致细菌性脑膜炎的病原。结论M-PCR方法较常规的分离培养鉴定方法具有较高的灵敏度和检测快速的特点,适合临床疑似细菌性脑膜炎感染病例的检测及筛查,提高临床细菌性感染脑膜炎病例的诊断阳性率。
Objective To establish a method for the detection of eight kinds of bacterial meningitis by multiplex polymerase chain reaction (M-PCR). Methods The species-specific genes of Neisseria meningitides, Haemophilus influenzae, Streptococcus pneumoniae, Staphylococcus aureus, Listeria monocytogenes, Streptococcus suis, Mycobacterium tuberculosis, Cryptococcus neoformans The single PCR amplification system and amplification program were optimized. M-PCR method was used to detect eight pathogens causing bacterial meningitis and the sensitivity of M-PCR system was tested. Results A preliminary M-PCR method for the detection of eight bacterial pathogens causing bacterial meningitis was initially established, allowing the simultaneous detection of a wide range of pathogens causing bacterial meningitis. Conclusion The M-PCR method has higher sensitivity and rapid detection than the conventional method for isolation and culture identification, and is suitable for the detection and screening of clinical bacterial infections with bacterial meningitis, and to improve the diagnostic positive rate of clinical bacterial meningitis .