Use of oligoprobes on amplified DNA in the diagnosis of bacterial meningitis

• Published in: European journal of clinical microbiology & infectious diseases Vol. 18; no. 5; pp. 352 - 357
• Main Authors: DICUONZO, G, LORINO, G, LILLI, D, RIVANERA, D, GUARINO, P, ANGELETTI, S, GHERARDI, G, CANDIDA, S, FILADORO, F
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01.05.1999; Springer Nature B.V
• Subjects: Bacteria; Bacterial diseases; Bacterial diseases of the nervous system. Bacterial myositis; Biological and medical sciences; Cerebrospinal fluid; DNA Primers; DNA Probes; E coli; Genomes; Glucose - cerebrospinal fluid; Haemophilus influenzae; Human bacterial diseases; Humans; Hybridization; Infectious diseases; Laboratories; Leukocyte Count; Medical sciences; Meningitis; Meningitis, Bacterial - cerebrospinal fluid; Meningitis, Bacterial - classification; Meningitis, Bacterial - diagnosis; Meningitis, Bacterial - microbiology; Neisseria meningitidis; Oligonucleotide Probes; Polymerase chain reaction; Polymerase Chain Reaction - methods; Sensitivity and Specificity; Streptococcus; Streptococcus infections; Tuberculosis; Rome Italy; Italy; Germany; Performance evaluation; Molecular hybridization; Neisseriaceae; Ribosomal DNA; Haemophilus influenzae; Specificity; Bacteria; Micrococcales; Meningitis; Molecular probe; Diagnosis; Human; Nervous system diseases; Neisseria meningitidis; Streptococcus pneumoniae; Infection; Pasteurellaceae; Polymerase chain reaction; Streptococcaceae; Gene amplification; Sensitivity; Mycobacterium tuberculosis; Mycobacteriales; Central nervous system disease; Mycobacteriaceae; Bacteriosis; Actinomycetes; Technique; Molecular biology
• ISSN: 0934-9723; 1435-4373
• DOI: 10.1007/PL00015018

An approach based on the 16 S rDNA polymerase chain reaction (16S PCR) and oligoprobe hybridization was applied to 77 cerebrospinal fluid samples submitted to the clinical microbiology laboratory for culture. Broad-range 16S rDNA primers were selected in conserved regions of the gene. Oligoprobes specific for Neisseria meningitidis, Haemophilus influenzae, Streptococcus spp., and Mycobacterium tuberculosis were selected in specific variable regions of the amplified 600 base pairs (bp) in the 16S rDNA. None of the oligoprobes cross hybridized with DNA from the other bacteria or from common contaminants. There were no false-negative results in culture-positive cerebrospinal fluid samples. Ten cases of meningitis caused by bacteria other than the four probes were not identified by any of the four probes. In culture-negative cerebrospinal fluid samples with some abnormal chemical 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.