Abstract
BackgroundPurulent meningitis (PM) is a serious life-threatening infection of the central nervous system (CNS) by bacteria or fungi and associated with high mortality and high incidence of CNS sequelae in children. However, the conventional cerebrospinal fluid (CSF) culture method is time-consuming and has a low sensitivity.MethodsOur study developed a real-time PCR-based purulent meningitis-TaqMan array card (PM-TAC) that targeted 21 PM-related pathogens and could produce results within 3 h. Primers and probes were adapted from published sources possibly. The performance of them were evaluated and optimized and then they were spotted on TAC.ResultsThe PM-TAC showed a sensitivity and specificity of 95 and 96%, respectively. For all of the 21 targeted pathogens, the PM-TAC assay had a LOD ranging from 5 copies/reaction to 100 copies/reaction, an intra-assay variation of 0.07–4.45%, and an inter-assay variation of 0.11–6.81%. Of the 15 CSF samples collected from patients with PM after empiric antibiotic therapies, the positive rate was 53.3% (8/15) for our PM-TAC assay but was only 13.3% (2/15) for the CSF culture method. Of the 17 CSF samples showing negative CSF culture, the PM-TAC assay identified a case of Neisseria meningitidis infection. Furthermore, all of the 10 CSF samples from patients without CNS infection showed negative for the PM-TAC assay.ConclusionsOur PM-TAC assay also demonstrated that the pathogen loads in the CSF samples correlated with the severity of PM. Thus, the PM-TAC may be helpful to improve the prognosis of PM and clinical outcomes from antibiotic therapies.
Highlights
Purulent meningitis (PM) is a serious life-threatening infection of the central nervous system (CNS) by bacteria or fungi and associated with high mortality and high incidence of CNS sequelae in children
We evaluated the sensitivity and specificity of the purulent meningitis-TaqMan array card (PM-TaqMan array card (TAC)) using artificial cerebrospinal fluid (ACSF) spiked with the targeted pathogens
The PM-TAC in this study can detect 21 PM-associated pathogens in 3 h and showed a higher sensitivity than the cerebrospinal fluid (CSF) culture method for CSF samples collected after empiric antibiotic therapies
Summary
Purulent meningitis (PM) is a serious life-threatening infection of the central nervous system (CNS) by bacteria or fungi and associated with high mortality and high incidence of CNS sequelae in children. The conventional cerebrospinal fluid (CSF) culture method is time-consuming and has a low sensitivity. Purulent meningitis (PM), which is often caused by bacterial or fungal infection, is a serious disease of the central nervous system and life-threatening for children and newborns. The direct Gram staining method can provide results rapidly and confirm a diagnosis of bacterial infection in 60–90% of patients, and a confirmed diagnosis from Gram staining depends on the bacterial concentration in CSF samples [7]. The Gram staining method may fail when the bacterial concentration is too low in CSF samples, and the sensitivity of the Gram staining method varies widely for different microorganisms [10]. Luminex Verigene, a high-throughput platform, can detect multiple pathogens from one sample simultaneously but requires a relatively long time to collect results [17]. The system is very expensive ($305–$453/sample and $22–$32/pathogen) [20, 21]
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