Establishment and evaluation of PCR-array for detection of six common pathogens of purulent meningitis in children / 军事医学

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.