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12 January 2018 Evaluation of Two Multiplex PCR–High-Resolution Melt Curve Analysis Methods for Differentiation of Campylobacter jejuni and Campylobacter coli Intraspecies
Banya Banowary, Van Tuan Dang, Subir Sarker, Joanne H. Connolly, Jeremy Chenu, Peter Groves, Shane Raidal, Seyed Ali Ghorashi
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Abstract

Campylobacter infection is a common cause of bacterial gastroenteritis in humans and remains a significant global public health issue. The capability of two multiplex PCR (mPCR)–high-resolution melt (HRM) curve analysis methods (i.e., mPCR1-HRM and mPCR2-HRM) to detect and differentiate 24 poultry isolates and three reference strains of Campylobacter jejuni and Campylobacter coli was investigated. Campylobacter jejuni and C. coli were successfully differentiated in both assays, but the differentiation power of mPCR2-HRM targeting the cadF gene was found superior to that of mPCR1-HRM targeting the gpsA gene or a hypothetical protein gene. However, higher intraspecies 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.

Banya Banowary, Van Tuan Dang, Subir Sarker, Joanne H. Connolly, Jeremy Chenu, Peter Groves, Shane Raidal, and Seyed Ali Ghorashi "Evaluation of Two Multiplex PCR–High-Resolution Melt Curve Analysis Methods for Differentiation of Campylobacter jejuni and Campylobacter coli Intraspecies," Avian Diseases 62(1), 86-93, (12 January 2018). https://doi.org/10.1637/11739-080417-Reg.1
Received: 17 August 2017; Accepted: 1 January 2018; Published: 12 January 2018
KEYWORDS
Campylobacter coli
Campylobacter jejuni
differentiation
high-resolution melt curve analysis
multiplex PCR
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