A novel strategy for the isolation and identification of environmental Burkholderia cepacia complex bacteria.
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 microgml(-1) C-390, 600U ml(-1) polymyxin B sulfate, 10 microgml(-1) gentamycin, 2 microgml(-1) vancomycin and 10 microgml(-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 degrees 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 HaeIII-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.
Laboratory of Microbiology, Ghent University, Ledeganckstraat 35, B-9000 Ghent, Belgium. Elke.Vanlaere@Ugent.be
Elke
E
Vanlaere
Tom
T
Coenye
Emly
E
Samyn
Caroline
C
Van den Plas
John
J
Govan
Frans
F
De Baets
Kris
K
De Boeck
Christiane
C
Knoop
Peter
P
Vandamme
D019121:Burkholderia Infections / Q000209:etiology / Q000382:microbiology; D016956:Burkholderia cepacia / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D003550:Cystic Fibrosis / Q000382:microbiology; D006801:Humans; D007202:Indicators and Reagents; D012150:Polymorphism, Restriction Fragment Length; D011112:Polymyxin B; D012988:Soil Microbiology
D007202:Indicators and Reagents; D011112:Polymyxin B