Development and validation of a novel multiplex digital PCR assay for identification of pathogens in cerebrospinal fluid of children with bacterial meningitis

•A novel digital PCR assay was developed and validated to detect the eight pathogens most commonly responsible for bacterial meningitis.•Digital PCR has a better ability to detect pathogens compared to bacterial culture in bacterial meningitis, especially for Streptococcus pneumoniae.•A high bacteri...

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Bibliographic Details
Published in:Clinica chimica acta 2024-02, Vol.554, p.117787-117787, Article 117787
Main Authors: Yang, Ying, Hu, Yiting, Yang, Cheng, Shi, Wen, Jin, Sufeng, Hua, Chunzhen, Jiang, Kewen
Format: Article
Language:English
Subjects:
Bacterial meningitis
Children
digital PCR
Etiological diagnosis
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Summary:•A novel digital PCR assay was developed and validated to detect the eight pathogens most commonly responsible for bacterial meningitis.•Digital PCR has a better ability to detect pathogens compared to bacterial culture in bacterial meningitis, especially for Streptococcus pneumoniae.•A high bacterial DNA load identified by digital PCR may indicate poor outcomes.•The novel digital PCR assay may be a promising, supplementary method for the diagnosis of pathogens in children with bacterial meningitis. Identifying the pathogens of bacterial meningitis (BM) is crucial for its diagnosis and treatment. The aim of this study is to develop and validate a novel method for detecting pathogens in cerebrospinal fluid (CSF) of children with BM using a digital polymerase chain reaction (dPCR) assay.Materials and methods:A novel multiplex dPCR assay method has been developed and validated. The diagnostic performance of the dPCR assay was compared with that of synchronous CSF culture, and the factors affecting its performance were analyzed.Results:A total of 69 children with BM were enrolled prospectively. The sensitivity of the dPCR assay was 94.44 %, specificity was 100 %, coincidence rate was 98.55 %, Kappa value was 0.959, and net reclassification improvement was 61.11 %. Compared with the CSF culture assay, the dPCR assay had higher sensitivity in different bacterial groups. Multiple factors affected its performance, including previous use of antibiotics, sampling time, BM complications, and levels of inflammatory biomarkers in CSF and blood (all P 
ISSN:0009-8981
1873-3492
DOI:10.1016/j.cca.2024.117787