Differentiation between pathogenic and non-pathogenic Yersinia enterocolitica strains by colony hybridization with a PCR-mediated digoxigenin-dUTP-labelled probe.
The Polymerase Chain Reaction (PCR) method was used to generate a vector-free digoxigenin-dUTP labelled probe that targets the Yersinia enterocolitica gene encoding the heat stable enterotoxin (yst). The probe was used in DNA-DNA colony hybridization to screen 113 strains of Y. enterocolitica and related species for the presence of the enterotoxin gene. In Y. enterocolitica, the probe clearly discriminated between pathogenic and non-pathogenic strains even those belonging to the same serotype. Of the other Yersinia species, only three strains of Y. kristensenii possessed DNA sequences homologous to the yst gene. The probe was further checked for its specificity in artificially inoculated fecal samples and could easily detect the target sequence of the yst gene. The digoxigenin-labelled probe proved to be a reliable epidemiological tool to discriminate between pathogenic and non-pathogenic strains in pure and mixed culture, thus offering the advantage of using a non-radioactive detection system in clinical laboratories with the possibility of reusing the same hybridization solution several times and obtaining results within a relatively short time.
Ibrahim
A
A
Department of Microbiology, University of Queensland, Australia.
Liesack
W
W
Stackebrandt
E
E
D000818:Animals; D001427:Bacterial Toxins / Q000235:genetics*; D001483:Base Sequence; D015342:DNA Probes; D003856:Deoxyuracil Nucleotides; D004076:Digoxigenin / Q000031:analogs & derivatives; D004768:Enterotoxins / Q000235:genetics*; D005243:Feces / Q000382:microbiology; D006761:Hospitals; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D015007:Yersinia / Q000302:isolation & purification; D015008:Yersinia enterocolitica / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity*
D001427:Bacterial Toxins; D015342:DNA Probes; D003856:Deoxyuracil Nucleotides; D004768:Enterotoxins; C070202:digoxigenin-11-deoxyuridine triphosphate; C048669:heat-stable enterotoxin (Yersinia); D004076:Digoxigenin