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

Four polymerase chain reaction (PCR) primer pairs (A-D), specific for the B urkholderia cepacia complex of organisms were designed to encompass the ent ire gene (similar to 1300 bp) from sequence alignments of the recA operon o f B. cepacia. Genomic bacterial DNA from type strains and wild-type B. cepa cia complex isolates of previously determined genomovar status was amplifie d employing these four primer pairs, as well as a fifth primer set (E) alre ady published. Primer sets B, C and E were successful in obtaining a PCR am plicon of correct estimated size of 598, 1107 and 1043 bp, respectively. Su bsequent single-stranded conformational polymorphism (SSCP) analysis of PCR amplicons demonstrated unique profiles for the five genomovar types for pr imer pairs B and E, but was unable to differentiate distinguishable profile types for primer pair C. SSCP analysis was demonstrated to be simple, rapi d and cost effective and may prove a useful method of genomovar typing of B . cepacia complex organisms from cystic fibrosis patients in busy diagnosti c laboratories. (C) 2001 The Hospital Infection Society.