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Diagnosis of meningococcal meningitis by broad-range bacterial PCR with cerebrospinal fluid
P Kotilainen1, J Jalava, O Meurman
1Department of Medicine, Turku University Central Hospital, Finland. pirkko.kotilainen@utu.fi
Abstract:
We used broad-range bacterial PCR combined with DNA sequencing to examine prospectively cerebrospinal fluid (CSF) samples from patients with suspected meningitis. Fifty-six CSF samples from 46 patients were studied during the year 1995. Genes coding for bacterial 16S and/or 23S rRNA genes could be amplified from the CSF samples from five patients with a clinical picture consistent with acute bacterial meningitis. For these patients, the sequenced PCR product shared 98.3 to 100% homology with the Neisseria meningitidis sequence. For one patient, the diagnosis was initially made by PCR alone. Of the remaining 51 CSF samples, for 50 (98.0%) samples the negative PCR findings were in accordance with the negative findings by bacterial culture and Gram staining, as well as with the eventual clinical diagnosis for the patient. However, the PCR test failed to detect the bacterial rRNA gene in one CSF sample, the culture of which yielded Listeria monocytogenes. These results invite new research efforts to be focused on the application of PCR with broad-range bacterial primers to improve the etiologic diagnosis of bacterial meningitis. In a clinical setting, Gram staining and bacterial culture still remain the cornerstones of diagnosis.
Insights
Broad-range bacterial PCR shows promise for diagnosing meningitis by detecting bacterial 16S rRNA genes in cerebrospinal fluid (CSF). While effective in identifying Neisseria meningitidis, further research is needed to improve detection rates for all bacterial meningitis cases.
Area of Science:
- Microbiology
- Molecular Biology
- Clinical Diagnostics
Background:
- Bacterial meningitis remains a significant cause of morbidity and mortality.
- Accurate and rapid etiologic diagnosis is crucial for effective treatment.
- Conventional diagnostic methods like Gram staining and culture have limitations.
Purpose of the Study:
- To evaluate the utility of broad-range bacterial PCR and DNA sequencing for diagnosing bacterial meningitis.
- To assess the performance of PCR in detecting bacterial rRNA genes in cerebrospinal fluid (CSF) samples.
- To compare PCR results with conventional diagnostic methods.
Main Methods:
- Prospective analysis of 56 CSF samples from 46 patients with suspected meningitis.
- Broad-range bacterial PCR targeting 16S and/or 23S rRNA genes.
- DNA sequencing of PCR products for bacterial identification.
- Comparison with Gram staining and bacterial culture results.
Main Results:
- Bacterial rRNA genes were amplified from 5 patients with acute bacterial meningitis, with sequences showing high homology to Neisseria meningitidis.
- PCR provided the initial diagnosis for one patient.
- PCR results were consistent with culture and clinical diagnosis in 50 out of 51 negative samples.
- PCR failed to detect Listeria monocytogenes in one culture-positive sample.
Conclusions:
- Broad-range bacterial PCR is a valuable tool for the etiologic diagnosis of bacterial meningitis, particularly for identifying Neisseria meningitidis.
- PCR can potentially expedite diagnosis, as demonstrated in one case.
- Further research is warranted to optimize PCR protocols for broader bacterial coverage and improve diagnostic accuracy.
- Gram staining and bacterial culture remain essential components of meningitis diagnosis in clinical practice.