军事医学2017，Vol.41Issue(11)：902-905,4.DOI:10.7644/j.issn.1674-9960.2017.11.008
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
Establishment and evaluation of PCR-array for detection of six common pathogens of purulent meningitis in children
摘要
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.关键词
化脓性脑膜炎病原菌/PCR阵列/实时荧光定量PCR/脑脊液Key words
pathogens of purulent meningitis/PCR-array/real-time fluorescence quantitative PCR/cerebrospinal fluid分类
医药卫生引用本文复制引用
赵成娜,张超,郑玉玲,刘鹏,姜永强..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[J].军事医学,2017,41(11):902-905,4.基金项目
国家自然科学基金资助项目(81772144) （81772144）
北京市自然科学基金资助项目(7172155) （7172155）
