Characterization of Bilophila wadsworthia isolates using PCR fingerprinting

Bilophila wadsworthia, an under-appreciated anaerobic organism, was origina lly described in 1989. Ninety-nine Bilophila wadsworthia isolates, recovere d form environmental and clinical specimens in Germany and in Southern Cali fornia, were examined in this study. Many isolates were recovered in mixed culture with facultative aerobic and other anaerobic bacteria. All isolates were identified by standard laboratory procedures, including gas-liquid ch romatography (GLC). A PCR fingerprint assay was established to compare the profiles of clinical and environmental isolates to the type strain (ATCC 49 260) and to an environmental (sewage) reference strain (DSM 11045, RZATAU) for intra-species differences. Two primers, one universal primer, M13 core, and one tDNA primer, T3B, were used individually to analyse the strains. H omogeneous PCR fingerprint profiles were found for the majority of strains using the M13 core primer; two PCR groups were determined with T3B, one mat ching the type strain and one matching the environmental reference strain ( DSM 11045, RZATAU). Two urease negative strains, WAL 11470 (blood isolate f rom California) and TUB 754 (intra-abdominal isolate from Germany) formed u nique PCR fingerprint profiles with each of these primers. These results we re confirmed by PCR fingerprinting using the T3A primer. These latter resul ts suggest a possible genetic diversity in B. wadsworthia. (C) 1999 Academi c Press.