title: Identification of aquatic Burkholderia (Pseudomonas) cepacia by hybridization with species-specific rRNA gene probes.
abstract: Burkholderia (Pseudomonas) cepacia is a common environmental bacterium which can be pathogenic for plants and humans. In this study, four strategies were used to identify aquatic isolates: API test strips, hybridization with species-specific DNA probes for the 16S and 23S rRNA genes, fatty acid methyl ester (FAME) profiles, and growth on selective medium (TB-T agar [C. Hagedorn, W. D. Gould, T. R. Bardinelli, and D. R. Gustarson, Appl. Environ. Microbiol. 53:2265-2268, 1987]). Only 59% of the isolates identified as B. cepacia with the API test strips were confirmed as B. cepacia by using fatty acid profiles. The 23S rRNA probe generated a few false-positive results but dramatically underestimated the number of B. cepacia isolates (i.e., 40% of the colonies that did not hybridize to the probe were B. cepacia, as determined by FAME). The 16S rRNA probe generated more false-positive results than the 23S rRNA probe but was effective in identifying the majority of the B. cepacia isolates. The selective medium was only partially successful in recovering B. cepacia. Use of the B. cepacia-specific 16S rRNA probe was the most efficient and accurate way of identifying B. cepacia.
authors.affiliation: Department of Microbiology and Savannah River Ecology Laboratory, University of Georgia, Athens 30602, USA.
authors.forename: L G
authors.initials: LG
authors.lastname: Leff
authors.forename: R M
authors.initials: RM
authors.lastname: Kernan
authors.forename: J V
authors.initials: JV
authors.lastname: McArthur
authors.forename: L J
authors.initials: LJ
authors.lastname: Shimkets
mesh_terms: D001483:Base Sequence; D016956:Burkholderia cepacia / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D015342:DNA Probes / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D012329:RNA, Bacterial / Q000235:genetics*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D012338:RNA, Ribosomal, 23S / Q000235:genetics*; D013045:Species Specificity; D014871:Water Microbiology
chemical_list: D015342:DNA Probes; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S