[Establishment and application of TaqMan Real-Time PCR in detection and serogrouping of Neisseria meningitidis].
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. 121 cerebrospinal fluid (CSF) specimens from suspected N. meningitidis invasive meningitis cases were detected by latex agglutination test and Real-Time PCR assay simultaneously.


RESULTS
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 by 10(1)-10(3) times. The respective sensitivities for ctrA, sacB, siaD (B), siaD (C), xcbB, synF and synG were 8, 8, 80, 8, 8, 80, 8 genome DNA copies in each reaction. Of the 121 CSF specimens, 11 were positive for Real-Time PCR and 6 for latex agglutination test.


CONCLUSIONS
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 by N. meningitidis.
State Key Laboratory for Infectious Disease Control and Prevention, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
Bing-Qing
BQ
Zhu
Li
L
Xu
Ma-Chao
MC
Li
Hong-Yu
HY
Ren
Guo-Zhong
GZ
Tian
Yuan
Y
Gao
Yan-Hua
YH
Wang
Guo-Ming
GM
Qi
Biao
B
Kan
Zhi-Jun
ZJ
Shao
D009345:Neisseria meningitidis / Q000145:classification* / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012703:Serotyping / Q000379:methods*