Evaluation of two multiplex PCR–High-Resolution melt curve analysis methods for differentiation of Campylobacter jejuni and Campylobacter coli intraspecies

Campylobacter infection is a common cause of bacterial gastroenteritis in humans and remains a significant global public health issue. Capability of two multiplex-PCR-HRM curve analysis methods to detect and differentiate 24 poultry isolates and three reference strains of C. jejuni and C. coli was investigated. Campylobacter jejuni and C. coli were successfully differentiated in both assays, while the differentiation power of mPCR2-HRM targeting the cadF gene was found superior to that of mPCR1-HRM targeting the gpsA or a hypothetical protein gene. However, higher intra-species variation within C. coli and C. jejuni isolates was detected in mPCR1-HRM when compared with mPCR2-HRM. Both assays were rapid and required minimum interpretation skills for discrimination between and within Campylobacter species when using HRM curve analysis software.