To develop a rapid detection method for Yersinia enterocolitica, dot blot hybridization with nonradioactively digoxigenin-labeled DNA probe was performed. Sixty-five Y. enterocolitica stratins and the other twenty-six Yersinia species were
collected in
Korea and around the world. Polymerase chain reaction(PCR) was performed with two 21-mer oligonucleotide primers against the ail gene of pathogenic Y. enterocolitica. PCR amplified 458bp DNA fragment, which was then used as a digoxigenin-labeled
probe.
We hybridized the ail gene probe with ninety-one chromosomal DNA of Yersinia species. Out of 65 Y. entercolitica strains, 55 strains hybridized with the ail gene probe and classified as pathogens, while 10 stratins did not. Thirteen strains of Y.
pseudotuberculosis, 4 strains of Y. kristensenii, 4 strains of Y. intermedia, 3 strains of Y. frederiksenii, 1 strain of Y. mollaretii, and 1 strain of Y. ruckeri were all negative with the ail gene probe. This result showed that the
digoxigenin-labeled
ail gene probe had 100% specificity.
|