Towards accurate exclusion of neonatal bacterial meningitis: a feasibility study of a novel 16S rDNA PCR assay

Autor:	Arthur Abelian, Carol Charan, Amanda Ogilvy-Stuart, Thomas Mund, Martin D. Curran, Paul H. Dear, Nipa Mitra, Stuart A. Savill, Hugh R.B. Pelham
Jazyk:	angličtina
Rok vydání:	2020
Předmět:	medicine.medical_specialty Microbiological culture 16s rdna pcr Neonatal bacterial meningitis DNA Ribosomal Polymerase Chain Reaction Sensitivity and Specificity Infant Newborn Diseases Microbiology Meningitis Bacterial lcsh:Infectious and parasitic diseases 03 medical and health sciences 0302 clinical medicine Medical microbiology 030225 pediatrics medicine Humans lcsh:RC109-216 030212 general & internal medicine Cerebrospinal Fluid Bacteria business.industry Infant Newborn Bactericidal effect medicine.disease Infectious Diseases Parasitology Broad-range PCR Ethidium monoazide Feasibility Studies Bacterial meningitis business Meningitis Field conditions Research Article
Zdroj:	BMC Infectious Diseases, Vol 20, Iss 1, Pp 1-11 (2020) BMC Infectious Diseases
ISSN:	1471-2334
DOI:	10.1186/s12879-020-05160-x
Popis:	BackgroundPCRctic is an innovative assay based on 16S rDNA PCR technology that has been designed to detect a single intact bacterium in a specimen of cerebro-spinal fluid (CSF). The assay’s potential for accurate, fast and inexpensive discrimination of bacteria-free CSF makes it an ideal adjunct for confident exclusion of bacterial meningitis in newborn babies where the negative predictive value of bacterial culture is poor. This study aimed to stress-test and optimize PCRctic in the “field conditions” to attain a clinically useful level of specificity.MethodsThe specificity of PCRctic was evaluated in CSF obtained from newborn babies investigated for meningitis on a tertiary neonatal unit. Following an interim analysis, the method of skin antisepsis was changed to increase bactericidal effect, and snap-top tubes (Eppendorf™) replaced standard universal containers for collection of CSF to reduce environmental contamination.ResultsThe assay’s specificity was 90.5% in CSF collected into the snap-top tubes – up from 60% in CSF in the universal containers. The method of skin antisepsis had no effect on the specificity. All CSF cultures were negative and no clinical cases of neonatal bacterial meningitis occurred during the study.ConclusionsA simple and inexpensive optimization of CSF collection resulted in a high specificity output. The low prevalence of neonatal bacterial meningitis means that a large multi-centre study will be required to validate the assay’s sensitivity and its negative predictive value.
Databáze:	OpenAIRE
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