Rapid characterization of the genomovars of the Burkholderia cepacia complex by PCR-single-stranded conformational polymorphism (PCR-SSCP) analysis.
Four polymerase chain reaction (PCR) primer pairs (A-D), specific for the Burkholderia cepacia complex of organisms were designed to encompass the entire gene (approximately 1300 bp) 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 1043 bp, 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.
Moore
J E
JE
Department of Bacteriology, Northern Ireland Public Health Laboratory, Belfast, BT9 7AD, UK. jemoore@niphl.dnet.co.uk
Millar
B C
BC
Jiru
X
X
McCappin
J
J
Crowe
M
M
Elborn
J S
JS
D016956:Burkholderia cepacia / Q000145:classification / Q000235:genetics*; D003550:Cystic Fibrosis / Q000382:microbiology; D017931:DNA Primers / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D018807:Polymorphism, Single-Stranded Conformational; D011938:Rec A Recombinases / Q000235:genetics; D012680:Sensitivity and Specificity
D017931:DNA Primers; D011938:Rec A Recombinases