Submitted to: Meeting Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: 5/9/2007
Publication Date: 7/20/2007
Citation: Abdelbaqi, K., Buissonniere, A., Prouzet-Mauleon, V., Gresser, J., Wesley, I.V., Desmonts, M., Megraud, F., Menard, A. Development of a real-time fluorescence resonance energy transfer (fret) PCR to detect Arcobacter species [abstract]. Campylobacter Helicobacter and Related Organisms (CHRO) 2007. 45(9):3015-3021.
Technical Abstract: A real-time PCR targeting the gyrase A subunit gene outside the quinolone resistance-determining region has been developed to detect Arcobacter species. The species identification was made by probe hybridization and melting curve analysis, using the Fluorescence Resonance Energy Transfer technology. Discrimination between Arcobacter species was straightforward, as the corresponding melting points showed significant differences with the characteristic melting temperatures of 63.5°C, 58.4°C, 60.6°C and 51.8°C, for Arcobacter butzleri, Arcobacter cryaerophilus, Arcobacter cibarius, and Arcobacter nitrofigilis type strains, respectively. The specificity of this assay was confirmed with pure cultures of 106 Arcobacter isolates from human clinical and veterinary specimens, identified by phenotypic methods and 16S rRNA gene sequencing. The assay was then used to screen 345 clinical stool samples obtained from patients with diarrhea. The assay detected A. butzleri in four of these clinical samples (1.2%). These results were confirmed by a conventional PCR method targeting the 16S rRNA gene with subsequent sequencing of the PCR product. In conclusion, this real-time assay detects and differentiates Arcobacter species in pure culture as well as in the competing microbiota of the stool matrix. The assay is economical since only one biprobe is used and multiple Arcobacter species are identified in a single test.