Diagnosis of Meningococcal Meningitis by Broad-Range Bacterial PCR with Cerebrospinal Fluid

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.

Employment of broad-range 16S rRNA PCR to detect aetiological agents of infection from clinical specimens in patients with acute meningitis - rapid separation of 16S rRNA PCR amplicons without the need for cloning

“…It has been reported that PCR amplification of numerous targets, such as the 16S rRNA gene, the 16S-23S rRNA interspacer region, the IS1106 region and the PorA gene, have been used to detect organisms in CSF and blood specimens from patients with meningitis (Radström et al, 1994;Hall et al 1995;Kotilainen et al 1998;Glustein et al 1999;Rantakokko-Jalava et al 2000;Seward and Towner, 2000). Molecular-based diagnostic approaches are now more frequently used alongside conventional microbiological techniques in the diagnostic clinical microbiology laboratory.…”

Bacterial components in the synovial tissue of patients with advanced rheumatoid arthritis or osteoarthritis: Analysis with gas chromatography‐mass spectrometry and pan‐bacterial polymerase chain reaction

“…Moreover, the bacterial DNA was detected in SF of 33% of ReA patients (Figure 4), and was consistent with the previous observations (39, 40). The PCR primers used have also been tested in previous studies and shown to cover most bacteria (26, 36, 37). It should be noted that the pan‐bacterial PCR is extremely sensitive to contamination.…”

“…The primers for 23S and 16S rDNA PCR have been described before. Both primer pairs, MS‐37 and MS‐38 for the 23S PCR and fD1 and rP2 for 16S PCR, have been used for diagnosis of bacterial infection for a variety of clinical samples (26, 36, 37).…”

Quality assessed nonculture techniques for detection and typing of meningococci

“…Detection of N. meningitidis DNA may be performed by amplification of conserved genes such as the genes encoding 16S rRNA. This amplification may target specific or universal sequences within these genes, and further sequencing the PCR product allows the identification of N. meningitidis (Kotilainen et al, 1998;Bäckman et al, 1999;Corless et al, 2000;Mölling et al, 2002).…”