Cited by 234Bacterial Identification and Susceptibility TestingBacterial Infections and VaccinesInfectious Diseases and Tuberculosis

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Prospective Study of Use of PCR Amplification and Sequencing of 16S Ribosomal DNA from Cerebrospinal Fluid for Diagnosis of Bacterial Meningitis in a Clinical Setting

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Schuurman, Tim, de Boer, Richard F., Kooistra-Smid, Anna M. D., van Zwet, Anton A.·2004·Journal of Clinical Microbiology

DOI 10.1128/jcm.42.2.734-740.2004PMC PMC344470

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Abstract

We have evaluated the use of a broad-range PCR aimed at the 16S rRNA gene in detecting bacterial meningitis in a clinical setting. To achieve a uniform DNA extraction procedure for both gram-positive and gram-negative organisms, a combination of physical disruption (bead beating) and a silica-guanidiniumthiocyanate procedure was used for nucleic acid preparation. To diminish the risk of contamination as much as possible, we chose to amplify almost the entire 16S rRNA gene. The analytical sensitivity of the assay was approximately 1 x 10(2) to 2 x 10(2) CFU/ml of cerebrospinal fluid (CSF) for both gram-negative and gram-positive bacteria. In a prospective study of 227 CSF samples, broad-range PCR proved to be superior to conventional methods in detecting bacterial meningitis when antimicrobial therapy had already started. Overall, our assay showed a sensitivity of 86%, a specificity of 97%, a positive predictive value of 80%, and a negative predictive value of 98% compared to culture. We are currently adapting the standard procedures in our laboratory for detecting bacterial meningitis; broad-range 16S ribosomal DNA PCR detection is indicated when antimicrobial therapy has already started at time of lumbar puncture or when cultures remain negative, although the suspicion of bacterial meningitis remains.

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Authors: Schuurman, T., de Boer, R. F., Kooistra-Smid, A. M. D., & van Zwet, A. A.
Affiliations: Department of Research and Development, Regional Public Health Laboratory for Groningen and Drenthe, Groningen, The Netherlands
Corresponding author: Schuurman, Tim
ORCID: Kooistra-Smid, Anna M. D.
Venue: Journal of Clinical Microbiology
Publisher: American Society for Microbiology
Published: 2004-02
Volume: 42
Issue: 2
Pages: 734-740
ISSN: 0095-1137 (ISSN-L)
1098-660X
DOI: 10.1128/jcm.42.2.734-740.2004
PMC: PMC344470
OpenAlex: W2070801207
Cited by: 234
References: 63
Citations by year: 2026: 9 · 2025: 6 · 2024: 7 · 2023: 8 · 2022: 5 · 2021: 11 · 2020: 14 · 2019: 13 · 2018: 13 · 2017: 7 · 2016: 10 · 2015: 12 · 2014: 8 · 2013: 13 · 2012: 12
FWCI: 6.51
Type: journal article
Language: en
Open access: green
www.ncbi.nlm.nih.gov
Licence: journals.asm.org/non-commercial-tdm-license (tdm)
Locations: Journal of Clinical Microbiology (publishedVersion)
PubMed Central (publishedVersion)
PubMed (publishedVersion)
Topics: Health Sciences › Medicine › Microbiology (medical) › Bacterial Identification and Susceptibility Testing
Life Sciences › Immunology and Microbiology › Microbiology › Bacterial Infections and Vaccines
Health Sciences › Medicine › Infectious Diseases › Infectious Diseases and Tuberculosis
Keywords: bacterial meningitis, 16S rRNA gene, cerebrospinal fluid, broad-range PCR, molecular diagnosis, nucleic acid extraction, bead beating, guanidine thiocyanate, sensitivity and specificity, antimicrobial therapy, culture-negative meningitis
MeSH terms: Base Sequence, DNA, Ribosomal, Escherichia coli, Escherichia coli Infections, Gene Amplification, Humans, RNA, Ribosomal, 16S, Sensitivity and Specificity, Staphylococcal Infections, Reproducibility of Results, Polymerase Chain Reaction, Meningitis, Bacterial, DNA Primers

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Prospective Study of Use of PCR Amplification and Sequencing of 16S Ribosomal DNA from Cerebrospinal Fluid for Diagnosis of Bacterial Meningitis in a Clinical Setting

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