鉴别脑膜炎奈瑟菌A、B、C、Y、W<sub>135</sub>群的多重聚合酶链反应诊断方法

张力; 邵祝军; 徐丽

Abstract

张力,邵祝军,徐丽.鉴别脑膜炎奈瑟菌A、B、C、Y、W₁₃₅群的多重聚合酶链反应诊断方法[J].Chinese journal of Epidemiology,2006,27(5):399-401

鉴别脑膜炎奈瑟菌A、B、C、Y、W₁₃₅群的多重聚合酶链反应诊断方法

A novel method for detection and sero-grouping of Nesseria meningitidis

Received:January 10, 2005

DOI：

KeyWord: 脑膜炎奈瑟菌多重聚合酶链反应

English Key Word: Neningitides Multiplex polymerase chain reaction

FundProject:

Author Name	Affiliation
ZHANG Li	Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
SHAO Zhu-jun	Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
XU Li	Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China

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Abstract:

目的建立一种能鉴别脑膜炎奈瑟菌(Nm)A、B、C、Y、W135个血清群的多重聚合酶链反应(PCR)方法。方法首先PCR扩增Nm crgA基因,确定Nm感染;再通过多重PCR扩增荚膜表达基因orf-2和siaD基因,根据特异条带区分不同群Nm。结果多重PCR检测61株Nm与4株非Nm,能准确地鉴定并可将Nm分成5个血清群;检测4例流行性脑脊髓膜炎患者的脑脊液,2例出现A群400 bp的特异条带,而细菌培养和胶乳凝集方法检测均为阴性;检测18份流行性乙型脑炎患者标本,与胶乳凝集试验结果一致。结论多重PCR诊断方法能正确鉴别不同群Nm,对流行性脑脊髓膜炎的临床诊断与流行病学调查有重要意义。

English Abstract:

Objective We developed a multiplex polymerase chain reaction(PCR ) method for detection, identification and sero-grouping strains of N.mertingitidis.Methods The gene of crgA was selected for detection and identification of N. mertingitidis and the 230 by of crgA fragments were amplified. The genes of siaD and orf-2 were used for sero-grouping. With this technology, N.meningitides of serogroup A,B,C,Y and W,33 can be differentiated from the specific fragment 400 bp,450 6p,250 6p,120 6p,120 6p. Results This multiple PCR method seemed to be more sensitive than latex agglutination.We used this method to detect 61 isolates of N. mertingitidis and the 230 by of crgA fragments were amplified. No positive results were obtained from on 4 strains of non-N.meningitides.It seemed that the PCR method which was based on crag, was specific in detecting N.mertingitidis.The 55 strains of N. mertingitidis could be grouped into A,B,C, Y and W,}S by this multiplex I'CR. No gene fragment was synthesized for the other serogroups of N. mertingitidis.No cross-reaction was observed for other bacterial species. 4 samples of meningococcal meningitis patients were detected by the multiplex PCR and two of them were positive with 400 by band,representing serogroup A of N. mertingitidis but all were negative to culture and latex agglutination. 18 samples of Japanese(B ) encephalitis patients were also detected by multiplex PCR and no positive result was obtained which was consistent to the finding from culture and latex agglutination.Conclusion This multiplex PCR method showed its potential in clinical diagnosis and epidemiological investigation.

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