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

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 access
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
Open this paper in CoChat — free account

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.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1128/jcm.42.2.734-740.2004

Cites

63 of 63 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Previous studies indicate that 16S rRNA PCR/sequencing might be a useful addition in diagnosing bacterial CNS infections when species-specific PCR and cultures remain negative.

    “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]

    The Discussion reports that broad-range 16S rDNA PCR with sequencing is a valuable supplementary test when CSF cultures remain negative despite suspicion of bacterial meningitis. The Results also identify pathogens in six culture-negative samples. The paper does not establish its usefulness specifically when species-specific PCR is negative; it compares the method with culture and mentions species-specific PCR only in discussing contamination risk. (LLM, conf=0.85)

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 2

  1. [8]Busch, N., P. R. Mertens, T. Schonfelder, H. Nguyen, H. U. Marschall, H. Kierdorf, G. Haase, R. Urhahn, H. G. Sieberth, and S. Matern. 1996. Lemierre's post-tonsillitis sepsis with meningitis and intravascular consumption coagulopathy as complication of infectious mononucleosis with pansinusitis. Dtsch. Med. Wochenschr. 121:94-98. [DOI] [PubMed] [Google Scholar]
  2. [56]Vernazza, P. L., J. R. Dyer, S. A. Fiscus, J. J. Eron, and M. S. Cohen. 1997. HIV-1 viral load in blood, semen and saliva. AIDS 11:1058-1059. [PubMed] [Google Scholar]

Cited by

Sorted by citations

Related

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
Licence
journals.asm.org/non-commercial-tdm-license (tdm)
Locations
  • Journal of Clinical Microbiology (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Clinical Biochemistry › Bacterial Identification and Susceptibility Testing
  • Life Sciences › Immunology and Microbiology › Microbiology › Bacterial Infections and Vaccines
  • Health Sciences › Medicine › Surgery › 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

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review