| Autor: |
Shimoji, Yoshihiro, Mori, Yasuyuki, Hyakutake, Koji, Sekizaki, Tsutomu, Yokomizo, Yuichi |
| Zdroj: |
Journal of Clinical Microbiology; January 1998, Vol. 36 Issue: 1 p86-89, 4p |
| Abstrakt: |
ABSTRACTWe have previously described the creation by Tn916mutagenesis of avirulent transposition mutants from a highly virulent strain of Erysipelothrix rhusiopathiae, the causative agent of swine erysipelas. In this study, we cloned a 2.2-kb DNA fragment which flanked the Tn916insertion in an avirulent mutant (strain 33H6) and evaluated the possibility that this region could be used for the specific detection of E. rhusiopathiae. According to the sequences of this region, oligonucleotide primers were designed to amplify a 937-bp fragment of the E. rhusiopathiaechromosome by PCR. The specificity of the PCR was investigated by analyzing 64 strains of Erysipelothrixspecies and 27 strains of other genera different fromErysipelothrix. A 937-bp DNA fragment could be amplified from all E. rhusiopathiaestrains tested, and no amplification was observed by using DNAs from the other species tested. To make a rapid and definite diagnosis of swine erysipelas in slaughterhouses, we developed an enrichment broth cultivation-PCR combination assay, which used a commercially available DNA extraction kit, to identify E. rhusiopathiaein the specimens from swine with arthritis. After samples were enriched in selective broth culture, detection of E. rhusiopathiaewas tested by either conventional methods or the PCR. Of 102 samples tested, 15 samples were positive by conventional methods and 12 of the 15 samples were positive by the PCR. The detection limit of the PCR was 103CFU per reaction mixture for the PCR-positive samples. These results indicate that this PCR technique could be used as a first-line screening technique for the specific detection of E. rhusiopathiaein specimens. |
| Databáze: |
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