Evaluation of three oligonucleotide primer sets in PCR for the identification of Burkholderia cepacia and their differentiation from Burkholderia gladioli.
OBJECTIVE
To evaluate three oligonucleotide primer pairs--two specific for 16S and 23S rRNA sequences of Burkholderia cepacia, and the third specific for internal transcribed spacer region of 16S-23S sequences of B gladioli--for the identification and differentiation of reference and clinical strains of these and other species.


METHODS
The three primers sets were applied in polymerase chain reaction (PCR) to a collection of 177 clinical isolates submitted for identification from diagnostic laboratories as presumed B cepacia.


RESULTS
At an annealing temperature of 63 degrees C, all eight B cepacia and four B gladioli reference strains reacted with their specific primers. B vandii was the only other species that was positive with both B cepacia primers but five Burkholderia or Ralstonia species reacted with one of these primers. Seventy eight isolates were typical of B cepacia in biochemical tests and 75 of these reacted with specific primers; three, however, were positive with the B gladioli primers. Fifteen asaccharolytic isolates were confirmed as B cepacia by PCR but other non-fermenting Gram negative species were negative with each of the primers.


CONCLUSIONS
PCR using 16S rRNA sequences is recommended for identification of B cepacia that give atypical results in biochemical tests.
Clode
F E
FE
Laboratory of Hospital Infection, Central Public Health Laboratory, London, UK.
Kaufmann
M E
ME
Malnick
H
H
Pitt
T L
TL
D019117:Burkholderia / Q000235:genetics; D019121:Burkholderia Infections / Q000175:diagnosis* / Q000382:microbiology; D016956:Burkholderia cepacia / Q000235:genetics*; D003550:Cystic Fibrosis / Q000382:microbiology*; D017931:DNA Primers; D005069:Evaluation Studies as Topic; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000235:genetics; D012680:Sensitivity and Specificity
D017931:DNA Primers; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S