Establishment and application of TaqMan Real-Time PCR in detection and serogrouping of Neisseria meningitidis
10.3321/j.issn:0254-6450.2008.04.012
- VernacularTitle:TaqMan荧光定量PCR检测和鉴别不同血清群脑膜炎奈瑟菌方法的建立及应用
- Author:
Bing-Qing ZHU
1
;
Li XU
;
Ma-Chao LI
;
Hong-Yu REN
;
Guo-Zhong TIAN
;
Yuan GAO
;
Yan-Hua WANG
;
Guo-Ming QI
;
Biao KAN
;
Zhu-Jun SHAO
Author Information
1. 中国疾病预防控制中心传染病预防控制所
- Keywords:
Neisseria meningitidis;
TaqMan;
Real-Time PCR
- From:
Chinese Journal of Epidemiology
2008;29(4):360-364
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish TaqMan Real-Time PcR method for detection and identification of Neisseria meningitidis.Methods Seven sets of primers and FAM-labeled probes targeting different genes of Neisseria meningitidis were designed and synthesized.ctrA gene was used for identification of N.meningitidis species.Six serogruops(A,B,C,X,Y,W135)of N.meningitidis were detected with following genes:sacB(A),siaD(B),siaD(C),xcbB(X),synF(Y)and synG(W135)respectively.Sensitivity and specificity of Real-Time PCR were assessed for different primers and probes.121cerebrospinal fluid(CSF)specimens from suspected N.meningitidis invasive meningitis cases were detected by latex agglutination test and Real-Time PCR assay simultaneously.Resuits 79 N.meningitidis isolates of different serogroups could be detected and identified by seven sets of primers and probes in this study.Real-Time PCR seemed more sensitive than standard PCR bv 101-103 times.The respective sensitivities for ctrA,sacB,siaD(B),siaD(C),xcbB,synF and synG were 8,8,80,8,8,80,8 genomeDNA copies in each reaction.Of the 121 CSF specimens,11 were positive for Real-Time PCR and 6 for latex agglutination test.Conclusion Real-Time PCR could rapidly detect and identify N.meningitidis of different serogroups and seemed more sensitive.It could be widely used for diagnose of invasive meningitis caused bv N.meningitidis.
