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Development of a TaqMan Array card to target 21 purulent meningitis-related pathogens

BACKGROUND: 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. However, the conventional cerebrospinal fluid (CSF) culture method is time-consumin...

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Autores principales: Zhao, Chengna, Wang, Xi, Zhang, Chao, Liu, Bing, Jing, Hongbo, Ming, Lihua, Jiang, Hua, Zheng, Yuling, Liu, Peng, Liu, Gang, Jiang, Yongqiang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6438039/
https://www.ncbi.nlm.nih.gov/pubmed/30922257
http://dx.doi.org/10.1186/s12879-019-3856-z
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author Zhao, Chengna
Wang, Xi
Zhang, Chao
Liu, Bing
Jing, Hongbo
Ming, Lihua
Jiang, Hua
Zheng, Yuling
Liu, Peng
Liu, Gang
Jiang, Yongqiang
author_facet Zhao, Chengna
Wang, Xi
Zhang, Chao
Liu, Bing
Jing, Hongbo
Ming, Lihua
Jiang, Hua
Zheng, Yuling
Liu, Peng
Liu, Gang
Jiang, Yongqiang
author_sort Zhao, Chengna
collection PubMed
description BACKGROUND: 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. However, the conventional cerebrospinal fluid (CSF) culture method is time-consuming and has a low sensitivity. METHODS: Our 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. RESULTS: The 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. CONCLUSIONS: Our 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12879-019-3856-z) contains supplementary material, which is available to authorized users.
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spelling pubmed-64380392019-04-08 Development of a TaqMan Array card to target 21 purulent meningitis-related pathogens Zhao, Chengna Wang, Xi Zhang, Chao Liu, Bing Jing, Hongbo Ming, Lihua Jiang, Hua Zheng, Yuling Liu, Peng Liu, Gang Jiang, Yongqiang BMC Infect Dis Research Article BACKGROUND: 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. However, the conventional cerebrospinal fluid (CSF) culture method is time-consuming and has a low sensitivity. METHODS: Our 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. RESULTS: The 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. CONCLUSIONS: Our 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12879-019-3856-z) contains supplementary material, which is available to authorized users. BioMed Central 2019-03-28 /pmc/articles/PMC6438039/ /pubmed/30922257 http://dx.doi.org/10.1186/s12879-019-3856-z Text en © The Author(s). 2019 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Zhao, Chengna
Wang, Xi
Zhang, Chao
Liu, Bing
Jing, Hongbo
Ming, Lihua
Jiang, Hua
Zheng, Yuling
Liu, Peng
Liu, Gang
Jiang, Yongqiang
Development of a TaqMan Array card to target 21 purulent meningitis-related pathogens
title Development of a TaqMan Array card to target 21 purulent meningitis-related pathogens
title_full Development of a TaqMan Array card to target 21 purulent meningitis-related pathogens
title_fullStr Development of a TaqMan Array card to target 21 purulent meningitis-related pathogens
title_full_unstemmed Development of a TaqMan Array card to target 21 purulent meningitis-related pathogens
title_short Development of a TaqMan Array card to target 21 purulent meningitis-related pathogens
title_sort development of a taqman array card to target 21 purulent meningitis-related pathogens
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6438039/
https://www.ncbi.nlm.nih.gov/pubmed/30922257
http://dx.doi.org/10.1186/s12879-019-3856-z
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