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PCR-restriction fragment length polymorphism analysis for detection of point mutations associated with macrolide
Sylvie Vacher1, Armelle Ménard, Elisabeth Bernard
1Centre National de Référence des Helicobacters et Campylobacters, Laboratoire de Bactériologie, Hopital Pellegrin, Bordeaux, France.
Abstract:
A 23S rRNA gene fragment in domain V was sequenced from 30 clinical isolates of Campylobacter spp., including 22 resistant to macrolides. Two point mutations associated with erythromycin resistance were identified at positions 2074 and 2075 on the 23S rRNA gene (homologous to A2142C and A2143G mutations in Helicobacter pylori) in which an adenine residue is also replaced with a cytosine and a guanine residue, respectively. A combined PCR-restriction fragment length polymorphism technique was developed to detect these mutations by using the BsaI and BceAI enzymes.