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Vaccimonitor

Print version ISSN 1025-028XOn-line version ISSN 1025-0298

Abstract

CLIMENT, Yanet et al. Detection and identification by PCR of microorganisms causing bacterial meningocephalitis. Vaccimonitor [online]. 2002, vol.11, n.2, pp.1-5. ISSN 1025-028X.

Our work reports the use of genomic DNA primers previously purified from reference strains to detect and identify the microorganisms that cause bacterial meningocephalitis (BME) in 24 samples of cerebrospinal fluid (CSF). General and specific primers derived from the 16S rRNA (ribosomal RNA) gene were used. The general primers were amplified in all the bacterial species analyzed:Neisseria meningitidis, type b Haemophylus influenzae and Streptococcus sp. in the same 825 pb band, but the specific ones only amplified in the microorganism from which they were derived. Bands of 667, 478 and 253 pb were respectively obtained for N. meningitidis,H. influenzae and Streptococcus sp. Streptococcus agalactiae was also detected by PCR in which a 478 pb band was obtained. Upon evaluating the primers in clinical CSF samples, 100% detection of the BME causing microorganisms was observed. The specificity was 100%. The PCR products were analyzed by Hae III digestion, and correspondence found for the restriction patterns of the clinical samples and the reference strains.

Keywords : PCR; Bacterium; ribosomal RNA; gene; CSF; BME.

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