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 Connolly, Jeremy Chenu, Peter Groves, Shane Raidal, Seyed Ghorashi

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

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.
Original languageEnglish
Pages (from-to)86-93
Number of pages8
JournalAvian Diseases
Volume62
Issue number1
DOIs
Publication statusPublished - 2018

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Campylobacter coli
Campylobacter jejuni
Campylobacter Infections
campylobacteriosis
Campylobacter
Gene Targeting
Multiplex Polymerase Chain Reaction
gastroenteritis
Gastroenteritis
gene targeting
assays
Poultry
public health
poultry
Software
Public Health
methodology
Proteins
genes
proteins

Grant Number

  • 40825
  • 40702

Cite this

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title = "Evaluation of Two Multiplex PCR–High-Resolution Melt Curve Analysis Methods for Differentiation of Campylobacter jejuni and Campylobacter coli Intraspecies",
abstract = "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.",
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Evaluation of Two Multiplex PCR–High-Resolution Melt Curve Analysis Methods for Differentiation of Campylobacter jejuni and Campylobacter coli Intraspecies. / Banowary, banya; Dang, Van Tuan; Sarker, Subir; Connolly, Joanne; Chenu, Jeremy; Groves, Peter; Raidal, Shane; Ghorashi, Seyed.

In: Avian Diseases, Vol. 62, No. 1, 2018, p. 86-93.

Research output: Contribution to journalArticle

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AU - Banowary, banya

AU - Dang, Van Tuan

AU - Sarker, Subir

AU - Connolly, Joanne

AU - Chenu, Jeremy

AU - Groves, Peter

AU - Raidal, Shane

AU - Ghorashi, Seyed

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AB - 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.

KW - Campylobacter coli

KW - Campylobacter jejuni

KW - multiplex-PCR

KW - high-resolution melt curve analysis, differentiation

KW - Differentiation

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