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

doi:10.1637/11739-080417-Reg.1

Evaluation of Two Multiplex PCR-High-Resolution Melt Curve Analysis Methods for Differentiation of Campylobacter jejuni and Campylobacter coli Intraspecies

Avian Dis. 2018 Mar;62(1):86-93. doi: 10.1637/11739-080417-Reg.1.

Authors

Banya Banowary¹, Van Tuan Dang¹, Subir Sarker^{1

2}, Joanne H Connolly^{1

3}, Jeremy Chenu⁴, Peter Groves⁵, Shane Raidal^{1

3}, Seyed Ali Ghorashi^{1

3}

Affiliations

¹ A School of Animal and Veterinary Sciences, Charles Sturt University, Wagga Wagga, New South Wales, Australia 2678.
² C School of Life Sciences, La Trobe University, Melbourne, Victoria, Australia 3086.
³ B Graham Centre for Agricultural Innovation, New South Wales Department of Primary Industries and Charles Sturt University, Wagga Wagga, New South Wales, Australia 2678.
⁴ D Birling Avian Laboratories, Bringelly, New South Wales, Australia 2556.
⁵ E University of Sydney, Sydney, New South Wales, Australia 2006.

PMID: 29620472
DOI: 10.1637/11739-080417-Reg.1

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.

Keywords: Campylobacter coli; Campylobacter jejuni; differentiation; high-resolution melt curve analysis; multiplex PCR.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Campylobacter Infections / classification
Campylobacter Infections / microbiology
Campylobacter Infections / veterinary*
Campylobacter coli / isolation & purification*
Campylobacter jejuni / isolation & purification*
Chickens
Multiplex Polymerase Chain Reaction / methods
Multiplex Polymerase Chain Reaction / veterinary*
Poultry Diseases / classification*
Poultry Diseases / microbiology