Identification and differentiation of Campylobacter species by high-resolution melting curve analysis.

Abstract

Campylobacter jejuni and Campylobacter coli are the important food-born zoonotic pathogen, also are leading causes of human food borne illnesses worldwide. cadF gene is expressed in all C.jejuni and C.coli strains and mediates cell binding to the cell matrix protein, Fibronectin. High-resolution melting (HRM) analysis is emerging as an efficient and robust method for discriminating DNA sequence variants. The goal of this study was to apply HRM analysis to identification of C.jejuni and C.coli. A total of 100 samples were obtained from chicken in Kermanshah, Iran. HRM analysis based on cadF gene and Eva Green was developed to the identification of Campylobacter to the species level. Fifty-five of 100 samples were positive as campylobacter (7 C.jejuni, 29 C.coli, 16 mixes and 3 unknown). Minor variations were observed in melting point temperatures of C.coli or C.jejuni isolates and this variation can be used to the differentiation between C.coli or C.jejuni isolates. The results of this study indicated that HRM curve analysis can be a suitable technique and rapid technique for distinguishing between C.jejuni and C.coli isolates.