Rapid characterization of the genomovars of the Burkholderia cepacia complex by pcr-single-stranded conformational polymorphism (PCR-SSCP) analysis

Abstract Four polymerase chain reaction (PCR) primer pairs (A–D), specific for the Burkholderia cepacia complex of organisms were designed to encompass the entire gene (,1300bp) from sequence alignments of the rec A operon of B. cepacia . Genomic bacterial DNA from type strains and wild-type B. cepacia complex isolates of previously determined genomovar status was amplified employing these four primer pairs, as well as a fifth primer set (E) already published. Primer sets B, C and E were successful in obtaining a PCR amplicon of correct estimated size of 598, 1107 and 1043bp, respectively. Subsequent single-stranded conformational polymorphism (SSCP) analysis of PCR amplicons demonstrated unique profiles for the five genomovar types for primer pairs B and E, but was unable to differentiate distinguishable profile types for primer pair C. SSCP analysis was demonstrated to be simple, rapid and cost effective and may prove a useful method of genomovar typing of B. cepacia complex organisms from cystic fibrosis patients in busy diagnostic laboratories.