1887

Abstract

The presence of the cytolethal distending toxin B gene () was examined in eight sp. flexispira reference strains, ATCC 700114 and 12 Finnish porcine strains and canine flexispira isolates. Part of the gene was amplified by PCR with degenerate primers VAT2 and DHF1, cloned and sequenced. The presence/absence of the gene as determined by PCR was confirmed by Southern hybridization and toxin production by HeLa cell-line experiments. PCR amplification resulted in approximately 700 bp fragments from sp. flexispira taxa 2 (ATCC 49314), 3 (ATCC 49320) and 8 (ATCC 43880, ATCC 49308, ATCC 43879), from six canine isolates as well as from the control strains and . The hybridization patterns of III-, dIII- and I-digested chromosomal DNA confirmed the results of the PCR experiments. The -positive strains had effects ranging from weak to strong on HeLa cell cultures. PCR amplification from the reference strains sp. flexispira taxa 1 (ATCC 43968), 4 (ATCC 49310) and 5 (ATCC 43966) and (ATCC 700114), and also six of the Finnish strains, was unsuccessful. No toxic effect on HeLa cells was evident when bacterial suspensions of PCR-negative strains were used for toxicity assay. Our results are in accordance with previous observations that the gene is not present in all species. Further, the presence/absence of the gene in sp. flexispira strains was in accordance with recent taxonomic analysis of the same strains, which suggests that it could serve as a useful marker in taxonomy.

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2003-02-01
2024-03-29
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