PCR方法从脑脊液中检测脑膜炎奈瑟菌的实验

Detection of Neisseria meningitidis from Cerebrospinal Fluid by Polymerase Chain Reaction

[目的]探索快速、可靠的检测脑膜炎奈瑟菌的方法。[方法]以煮沸法裂解细菌获取PCR反应模板,或以基因组DNA纯化试剂盒提取DNA后,用PCR法扩增脑膜炎奈瑟菌crgA基因;对PCR产物进行DNA测序。扩增siaD(C)基因确认C血清群脑膜炎奈瑟菌。[结果]煮沸法和试剂盒提取DNA法均可从1份标本中PCR扩增crgA基因阳性,PCR产物DNA序列与NCBI GenBankcrgA基因(登录号AF190471)序列一致。PCR扩增siaD(C)基因确认为C血清群脑膜炎奈瑟菌。[结论]建立的从脑脊液中检测脑膜炎奈瑟菌的PCR方法,可以更快速、可靠地诊断流脑。

[Objective] To explore a quick and reliable detection method for Neisseria meningitidis. [Methods] Cerebrospinal fluid specimens were boiled or extracted DNA by the extraction kit and crgA gene was detected by Polymerase Chain Reaction （PCR）. PCR products were sequenced. To confirm C serogroup Neisseria meningitidis, siaD （C） gene was detected by PCR method. [Results] One specimen was PCR positive to crgA and siaD （C） gene. CrgA gene PCR products＇ DNA sequence was coincident with crgA gene sequence registered in NCBI Gen-Bank. [Conclusion] The PCR method can detect out Neisseria meningitidis more quickly and reliably.