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J Med Microbiol 55 (2006), 721-727; DOI: 10.1099/jmm.0.46457-0
© 2006 Society for General Microbiology
ISSN 1473-5644

Identification of Burkholderia cepacia complex pathogens by rapid-cycle PCR with fluorescent hybridization probes

Ralf-Peter Vonberg1, Susanne Häußler1,2, Peter Vandamme3 and Ivo Steinmetz1,4

1 Institute of Medical Microbiology and Hospital Epidemiology, Medical School Hannover, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany

2 Department of Cell Biology, German Research Center for Biotechnology, Braunschweig, Germany

3 Laboratorium voor Mikrobiologie, Universiteit Gent, Belgium

4 Friedrich Loeffler Institute of Medical Microbiology, Ernst Moritz Arndt University Greifswald, Germany

Correspondence
Ivo Steinmetz
steinmetz.ivo{at}uni-greifswald.de

Received 7 December 2005
Accepted 17 February 2006

Members of the Burkholderia cepacia complex are important bacterial pathogens in cystic fibrosis (CF) patients. The B. cepacia complex currently consists of nine genetic subgroups (genomovars) of different epidemiological relevance and possibly of different pathogenic potential in humans. In this study, a new approach was developed for the rapid identification of B. cepacia genomovar I, Burkholderia multivorans (genomovar II), Burkholderia cenocepacia (lineage III-A and III-B), Burkholderia stabilis (genomovar IV) and Burkholderia vietnamiensis (genomovar V), which cause the large majority of infections in CF patients. The method was based on the detection of differences in the recA gene sequence by using rapid-cycle PCR and genomovar-specific fluorescence resonance energy transfer (FRET) probes. The genomovar status of all 39 B. cepacia complex strains tested (genomovars I–V) was identified by melting-curve analysis. Each FRET probe produced a specific fluorescence signal only with the respective genomovar, and not with other B. cepacia complex strains and Burkholderia spp. The identification system was easy to handle and revealed B. cepacia complex genomovar I–V status from culture isolates within about 1 h.

Abbreviations: CF, cystic fibrosis; FRET, fluorescence resonance energy transfer.

The strains used in this study are listed in Supplementary Table S1 with the online version of this paper.






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