Use of Oligoprobes on Amplified DNA in the Diagnosis of Bacterial Meningitis

Abstract An approach based on the 16 S rDNA polymerase chain reaction (16SPCR) and oligoprobe hybridization was applied to 77 cerebrospinal fluid samplessubmitted to the clinical microbiology laboratory for culture. Broad-range 16SrDNA primers were selected in conserved regions of the gene. Oligoprobes specificfor Neisseria meningitidis, Haemophilus influenzae, Streptococcus spp., and Mycobacteriumtuberculosis were selected in specific variable regions of the amplified 600base pairs (bp) in the 16S rDNA. None of the oligoprobes cross hybridized withDNA from the other bacteria or from common contaminants. There were no falsenegativeresults in culture-positive cerebrospinal fluid samples. Ten cases of meningitiscaused by bacteria other than the four probes were not identified by any of thefour probes. In culture-negative cerebrospinal fluid samples with some abnormalchemical parameters, there were 14 amplicons – one of Haemophilus influenzae,three of Streptococcus spp., six of Mycobacterium tuberculosis, and four not identified– while in normal cerebrospinal fluid samples there were no amplicons