ArticleInternational journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases2026
A novel real-time PCR assay for the simultaneous detection of the four main causes of bacterial meningitis.
Article in International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
objectivesCurrent multiplex assays cannot detect all four World Health Organization priority pathogens for meningitis diagnosis (Neisseria meningitidis, Haemophilus influenzae, Streptococcus pneumoniae, and Streptococcus agalactiae). This work developed a novel real-time polymerase chain reaction (PCR) assay capable of simultaneously detecting these pathogens.
methodsForty-four DNA samples, including type cultures (NCTC and ATCC), were tested. Specific primers and probes targeting sodC, to detect N. meningitidis, dmsA for H. influenzae, SP2020 for S. pneumoniae, and cfb for S. agalactiae were evaluated in monoplex and multiplex. Standard curves were generated, and limits of detection (LLD), slope, intercept, and R² were determined. Sensitivity, specificity, and predictive values (positive predictive values/negative predictive values [PPV/NPV]) were assessed for both monoplex and multiplex assays.
resultsMonoplex and multiplex assays showed equivalent performance. Sensitivities were 100% for all targets; specificities ranged 91.7% to 100%, PPVs 72.7% to 100%, and NPVs 100%. The multiplex assay showed high efficiency and consistent amplification for each target gene. LLDs ranged from 24 (S. pneumoniae) to 66 (H. influenzae) genome copies/µl.
conclusionThe multiplex PCR assay showed good performance for rapid and accurate detection of meningitis-associated bacteria. This test has applications for improved diagnosis of meningitis in low- and middle-income countries, where laboratory confirmation of major pathogens such as H. influenzae, S. pneumoniae, N. meningitidis, and Group B Streptococcus remains limited. Field validation with clinical specimens is required before implementation.
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