Characterization of a Tn5382-like transposon containing the vanB2 gene cluster in a Clostridium strain isolated from human faeces

Objectives: During a hospital surveillance programme to detect VRE carriers, an anaerobic vancomycin-resistant bacterial strain CCRI-9842 containing a vanB gene was isolated from a human faecal specimen. In this study, we have characterized this strain and its vanB-containing element. Methods: Strain CCRI-9842 was characterized by 16S rDNA sequencing and susceptibility testing. PCR mapping and sequencing of the vanB-containing element, as well as plasmid extraction and mating experiments, were carried out to investigate the genetic basis of vancomycin resistance in this strain. Results: Strain CCRI-9842 was identified as a Clostridium species closely related to Clostridium bolteae (96.8% 16S rDNA identity). This strain was resistant to a high level of vancomycin (MIC of 256 mg/L), but was susceptible to teicoplanin and ampicillin. The complete sequence of the CCRI-9842 vanB gene exhibited 99.1% identity with that of vanB2. PCR mapping and sequencing showed that the genetic element carrying vanB2 was similar to transposon Tn5382/Tn1549. This Tn5382-like transposon forms circular intermediates and is flanked on the left and right ends by repeat sequences of at least 700 bp in the opposite direction. No plasmid was detected in this strain, suggesting that the Tn5382-like transposon was integrated into the chromosome. The vancomycin resistance was not transferable to enterococci. Conclusions: Our report shows for the first time the presence of a Tn5382-like transposon carrying vanB2 in a Clostridium species of the human intestinal flora. This suggests that the vanB2 Tn5382-like transposon is an important vector for the spread of vancomycin resistance in several bacterial species.