A novel strategy for the isolation and identification of environmentalBurkholderia cepaciacomplex bacteria

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Abstract
The purpose of this study was to develop a novel strategy for the isolation and identification of Burkholderia cepacia complex bacteria from the home environment of cystic fibrosis (CF) patients. Water and soil samples were enriched in a broth containing 0.1%l-arabinose, 0.1%l-threonine, and a mixture of selective agents including 1 μg ml−1 C-390, 600 U ml−1 polymyxin B sulfate, 10 μg ml−1 gentamycin, 2 μg ml−1 vancomycin and 10 μg ml−1 cycloheximide. On selective media (consisting of the same components as above plus 1.8% agar), several dilutions of the enrichment broth were inoculated and incubated for 5 days at 28°C. Isolates with different randomly amplified polymorphic DNA patterns were inoculated in Stewart's medium. Putative B. cepacia complex bacteria were confirmed by means of recA PCR and further identified by Hae III-recA restriction fragment length polymorphism analysis. Our results suggest that these organisms may be more widespread in the home environment than previously assumed and that plant associated soil and pond water may be reservoirs of B. cepacia complex infection in CF patients.