Rapid detection of point mutations in domain V of the 23S rRNA gene in erythromycin-resistant Campylobacter isolates by pyrosequencing.

Abstract

Campylobacter is one of the most common foodborne pathogens causing acute gastroenteritis in humans. Erythromycin, a macrolide antibiotic, is the first-choice treatment for Campylobacter infections, and failure to eradicate Campylobacter is usually due to macrolide resistance. The most important mechanism responsible for macrolide resistance in Campylobacter is mediated by point mutations at position 2074 or 2075 in the peptidyl-transferase region of domain V of the 23S rRNA gene. In this study, the minimum inhibitory concentrations of 58 Campylobacter isolates (C. jejuni: n = 37; C. coli: n = 21) obtained from chickens were measured by agar dilution. Isolates were subjected to both pyrosequencing and Sanger sequencing methods to detect the 2074 and 2075 point mutations and evaluate the efficacy of the pyrosequencing method. The A2075G mutation was found to be the predominant mutation associated with erythromycin resistance. Compared with traditional methods, pyrosequencing is a novel, rapid, low-cost, and quantitative technology for detecting erythromycin resistance in Campylobacter.