title: Successful application of enzyme-labeled oligonucleotide probe for rapid and accurate cholera diagnosis in a clinical laboratory.
abstract: Two cholera cases were diagnosed using an enzyme-labeled oligonucleotide probe (ELONP) hybridization test for detection of cholera toxin gene (ctx) in a clinical laboratory at Osaka Airport Quarantine Station. The ELONP test with suspicious colonies of Vibrio cholerae O1 grown on TCBS or Vibrio agar plates gave positive result for ctx within 3 hr. We also tried to apply the ELONP test for direct detection of ctx in their stool and their non-selective culture. Specimens from Case #1, which contained 5.9 x 10(5) CFU/g of V. cholerae O1 in the stool, cultured for 7-8 hr or longer in alkaline peptone water or Marine broth at 37C, became positive for ctx. On the other hand, specimens from Case #2, which contained 8.7 x 10(8) CFU/ml (of V. cholerae O1 in the stool), gave positive result in this stool itself without any further culture. These data suggest that the ELONP test provides successfully a more rapid and accurate means of identifying "toxigenic" V. cholerae O1 in a clinical laboratory.
authors.affiliation: Quarantine Station, Osaka Airport, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Miyagi
authors.forename: Y
authors.initials: Y
authors.lastname: Matsumoto
authors.forename: K
authors.initials: K
authors.lastname: Hayashi
authors.forename: M
authors.initials: M
authors.lastname: Yoh
authors.forename: K
authors.initials: K
authors.lastname: Yamamoto
authors.forename: T
authors.initials: T
authors.lastname: Honda
mesh_terms: D000328:Adult; D000469:Alkaline Phosphatase; D002771:Cholera / Q000175:diagnosis*; D002772:Cholera Toxin / Q000235:genetics*; D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000032:analysis*; D005243:Feces / Q000382:microbiology; D005260:Female; D005798:Genes, Bacterial; D006801:Humans; D008297:Male; D015336:Molecular Probe Techniques; D015345:Oligonucleotide Probes; D015203:Reproducibility of Results; D014734:Vibrio cholerae / Q000235:genetics / Q000302:isolation & purification
chemical_list: D004269:DNA, Bacterial; D015345:Oligonucleotide Probes; D002772:Cholera Toxin; D000469:Alkaline Phosphatase