Establishment and evaluation of PCR-array for detection of six common pathogens of purulent meningitis in children
10.7644/j.issn.1674-9960.2017.11.008
- VernacularTitle:检测6种常见儿童化脓性脑膜炎病原菌的PCR阵列方法的建立与评价
- Author:
Cheng-Na ZHAO
1
;
Chao ZHANG
;
Yu-Ling ZHENG
;
Peng LIU
;
Yong-Qiang JIANG
Author Information
1. 安徽医科大学
- Keywords:
pathogens of purulent meningitis;
PCR-array;
real-time fluorescence quantitative PCR;
cerebrospinal fluid
- From:Military Medical Sciences
2017;41(11):902-905
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop a PCR-array method for detecting common purulent meningitis pathogens including Streptococcus pneumoniae,Escherichia coli,Haemophilus influenzae type B,Neisseria meningitidis,S.agalactiae and Listeria monocytogenes in children.Methods The amplification efficiency,limit of detection (LOD) and cross-reactivity were validated with individually real-time PCR using genomic DNA of the six pathogenic bacteria.The sensitivity and specificity of the PCR-array method were evaluated using artificial cerebrospinal fluid(CSF),and the consistency between the PCR-array method and the golden method of CSF culture was evaluated using clinical samples.Results The primers and probes of the pathogens in PCR-array had high specificity,and there was no cross reaction between them.The LOD of the PCR-array method was 10 cfu/ml and very sensitive.The sensitivity and specificity of the PCR-array method could reach 95％ in the evaluation of artificial CSF,and had a good consistency with the clinical gold standard method.Conclusion The PCR-array method with high sensitivity and specificity can simultaneously detect six common pathogens in children with purulent meningitis at 2.5 h,which could provide reference for the diagnosis of purulent meningitis.
