| Autor: |
Abdelbaqi, Khalil, Buissonnie`re, Alice, Prouzet-Mauleon, Vale´rie, Gresser, Jessica, Wesley, Irene, Me´graud, Francis, Me´nard, Armelle |
| Zdroj: |
Journal of Clinical Microbiology; September 2007, Vol. 45 Issue: 9 p3015-3021, 7p |
| Abstrakt: |
ABSTRACTA real-time PCR targeting the gyrase A subunit gene outside the quinolone resistance-determining region has been developed to detect Arcobacterspecies. The species identification was done by probe hybridization and melting curve analysis, using fluorescence resonance energy transfer technology. Discrimination between Arcobacterspecies 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 the Arcobacter butzleri, Arcobacter cryaerophilus, Arcobacter cibarius, and Arcobacter nitrofigilistype strains, respectively. The specificity of this assay was confirmed with pure cultures of 106 Arcobacterisolates 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. butzleriin 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 Arcobacterspecies 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 Arcobacterspecies are identified in a single test. |
| Databáze: |
Supplemental Index |
| Externí odkaz: |
|
