Identification of Burkholderia cepacia complex pathogens by rapid-cycle PCR with fluorescent hybridization probes.
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AbstractMembers 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.
CitationJ. Med. Microbiol. 2006, 55(Pt 6):721-7
- DNA-Based diagnostic approaches for identification of Burkholderia cepacia complex, Burkholderia vietnamiensis, Burkholderia multivorans, Burkholderia stabilis, and Burkholderia cepacia genomovars I and III.
- Authors: Mahenthiralingam E, Bischof J, Byrne SK, Radomski C, Davies JE, Av-Gay Y, Vandamme P
- Issue date: 2000 Sep
- Evaluation of species-specific recA-based PCR tests for genomovar level identification within the Burkholderia cepacia complex.
- Authors: Vermis K, Coenye T, Mahenthiralingam E, Nelis HJ, Vandamme P
- Issue date: 2002 Nov
- PCR-based detection and identification of Burkholderia cepacia complex pathogens in sputum from cystic fibrosis patients.
- Authors: McDowell A, Mahenthiralingam E, Moore JE, Dunbar KE, Webb AK, Dodd ME, Martin SL, Millar BC, Scott CJ, Crowe M, Elborn JS
- Issue date: 2001 Dec
- Burkholderia cepacia complex: distribution of genomovars among isolates from the maize rhizosphere in Italy.
- Authors: Fiore A, Laevens S, Bevivino A, Dalmastri C, Tabacchioni S, Vandamme P, Chiarini L
- Issue date: 2001 Feb
- PCR-based identification and characterization of Burkholderia cepacia complex bacteria from clinical and environmental sources.
- Authors: Seo ST, Tsuchiya K
- Issue date: 2004