Use of a LightCycler gyrA mutation assay for identification of ciprofloxacin-resistant Campylobacter coli.
A fluorescence resonance energy transfer-based assay has been employed to detect mutations at the 86 codon in the DNA gyrase A (gyrA) gene in Campylobacter coli strains. These mutations were associated with ciprofloxacin resistance in strains isolated in Italy in 2000. The mutations in the gyrA gene were detected by real-time PCR amplification followed by hybridization with two fluorescent probes designed with sequences complementary to the wild-type C. coli gyrA gene. Mutation detection was performed by melting peak analysis of the probe-PCR product hybrid performed on a LightCycler (Roche Diagnostic). This gyrA mutation assay allows a rapid and reproducible screening method of ciprofloxacin-resistant C. coli strains.
Laboratory of Bacteriology and Medical Mycology, Istituto Superiore di Sanità, viale Regina Elena 299, 00161 Rome, Italy. alecara@iss.it
Alessandra
A
Carattoli
Anna
A
Dionisi
Ida
I
Luzzi
D000818:Animals; D000890:Anti-Infective Agents / Q000494:pharmacology*; D001483:Base Sequence; D017000:Campylobacter coli / Q000145:classification* / Q000187:drug effects / Q000201:enzymology / Q000235:genetics; D002939:Ciprofloxacin / Q000494:pharmacology*; D027081:DNA Gyrase / Q000235:genetics*; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D004735:Energy Transfer; D005456:Fluorescent Dyes; D006801:Humans; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013696:Temperature
D000890:Anti-Infective Agents; D005456:Fluorescent Dyes; D002939:Ciprofloxacin; D027081:DNA Gyrase