The DOI matches its Crossref record; 63 of the 63 references it cites were found in a registry or an index.
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
Open accessAbstract
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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“Previous studies have indicated that 16S rRNA PCR/sequencing might be a useful addition in diagnosing bacterial CNS infections when species-specific PCR and cultures remain negative (35, 36).”
From Molecular diagnostics in cerebrospinal fluid for the diagnosis of central nervous system infections (2024) · cites this paper as [35]
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Metadata
- 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
- 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
- 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
- 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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