Development of rRNA-based PCR assays for identification of Burkholderia cepacia complex isolates recovered from cystic fibrosis patients

PCR assays targeting rRNA genes were developed to identify species (genomov ars) within the Burkholderia cepacia complex. Each assay was tested with 17 7 bacterial isolates that also underwent taxonomic analysis by whole-cell p rotein profile. These isolates were from clinical and environmental sources and included 107 B. cepacia complex strains, 23 Burkholderia gladioli stra ins, 20 Ralstonia pickettii strains, 10 Pseudomonas aeruginosa strains, 8 S tenotrophomonas maltophilia strains, and 9 isolates belonging to nine other species, The sensitivity and specificity of the 16S rRNA-based assay for B urkholderia multivorans (genomovar II) were 100 and 99%, respectively; for Burkholderia vietnamiensis (genomovar V), sensitivity and specificity were 87 and 92%, respectively, An assay based on 16S and 23S rRNA gene analysis of B, cepacia ATCC 25416 (genomovar I) was useful in identifying genomovars I, III, and TV as a group (sensitivity, 100%, and specificity, 99%), Anoth er assay, designed to be specific at the genus level, identified all but on e of the Burkholderia and Ralstonia isolates tested (sensitivity, 99%, and specificity, 96%), The combined use of these assays offers a significant im provement over previously published PCR assays for B. cepacia.