Evaluation of high-resolution melt curve analysis for rapid differentiation of Campylobacter hepaticus from other species in birds.

Spotty liver disease (SLD) is a bacterial disease of chicken, causing mortalities and reduction in egg production, hence, contributing to economic loss in the poultry industry. The causative agent of SLD has only recently been identified as a novel Campylobacter species, Campylobacter hepaticus. Spe...

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Autores principales:	Petrina Young, Pol Tarce, Sadhana Adhikary, Joanne Connolly, Tim Crawshaw, Seyed A Ghorashi
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/576e383e69744f5c9e5852f3972f8a44
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spelling	oai:doaj.org-article:576e383e69744f5c9e5852f3972f8a442021-12-02T20:05:35ZEvaluation of high-resolution melt curve analysis for rapid differentiation of Campylobacter hepaticus from other species in birds.1932-620310.1371/journal.pone.0251328https://doaj.org/article/576e383e69744f5c9e5852f3972f8a442021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0251328https://doaj.org/toc/1932-6203Spotty liver disease (SLD) is a bacterial disease of chicken, causing mortalities and reduction in egg production, hence, contributing to economic loss in the poultry industry. The causative agent of SLD has only recently been identified as a novel Campylobacter species, Campylobacter hepaticus. Specific primers were designed from the hsp60 gene of Campylobacter hepaticus and PCR followed by high-resolution melt curve analysis was optimised to detect and differentiate three species of Campylobacter (Campylobacter coli, Campylobacter jejuni and Campylobacter hepaticus). The three Campylobacter species produced a distinct curve profile and was differentiated using HRM curve analysis. The potential of the PCR-HRM curve analysis was shown in the genotyping of 37 Campylobacter isolates from clinical specimens from poultry farms. PCR-HRM curve analysis of DNA extracts from bile samples or cultures from bile samples, were identified as Campylobacter hepaticus and confirmed by DNA sequencing. The DNA sequence analysis of selected samples from each of the three HRM distinctive curves patterns showed that each DNA sequence was associated with a unique melt profile. The potential of the PCR-HRM curve analysis in genotyping of Campylobacter species was also evaluated using faecal specimens from 100 wild birds. The results presented in this study indicate that PCR followed by HRM curve analysis provides a rapid and robust technique for genotyping of Campylobacter species using either bacterial cultures or clinical specimens.Petrina YoungPol TarceSadhana AdhikaryJoanne ConnollyTim CrawshawSeyed A GhorashiPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 5, p e0251328 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Petrina Young Pol Tarce Sadhana Adhikary Joanne Connolly Tim Crawshaw Seyed A Ghorashi Evaluation of high-resolution melt curve analysis for rapid differentiation of Campylobacter hepaticus from other species in birds.
description	Spotty liver disease (SLD) is a bacterial disease of chicken, causing mortalities and reduction in egg production, hence, contributing to economic loss in the poultry industry. The causative agent of SLD has only recently been identified as a novel Campylobacter species, Campylobacter hepaticus. Specific primers were designed from the hsp60 gene of Campylobacter hepaticus and PCR followed by high-resolution melt curve analysis was optimised to detect and differentiate three species of Campylobacter (Campylobacter coli, Campylobacter jejuni and Campylobacter hepaticus). The three Campylobacter species produced a distinct curve profile and was differentiated using HRM curve analysis. The potential of the PCR-HRM curve analysis was shown in the genotyping of 37 Campylobacter isolates from clinical specimens from poultry farms. PCR-HRM curve analysis of DNA extracts from bile samples or cultures from bile samples, were identified as Campylobacter hepaticus and confirmed by DNA sequencing. The DNA sequence analysis of selected samples from each of the three HRM distinctive curves patterns showed that each DNA sequence was associated with a unique melt profile. The potential of the PCR-HRM curve analysis in genotyping of Campylobacter species was also evaluated using faecal specimens from 100 wild birds. The results presented in this study indicate that PCR followed by HRM curve analysis provides a rapid and robust technique for genotyping of Campylobacter species using either bacterial cultures or clinical specimens.
format	article
author	Petrina Young Pol Tarce Sadhana Adhikary Joanne Connolly Tim Crawshaw Seyed A Ghorashi
author_facet	Petrina Young Pol Tarce Sadhana Adhikary Joanne Connolly Tim Crawshaw Seyed A Ghorashi
author_sort	Petrina Young
title	Evaluation of high-resolution melt curve analysis for rapid differentiation of Campylobacter hepaticus from other species in birds.
title_short	Evaluation of high-resolution melt curve analysis for rapid differentiation of Campylobacter hepaticus from other species in birds.
title_full	Evaluation of high-resolution melt curve analysis for rapid differentiation of Campylobacter hepaticus from other species in birds.
title_fullStr	Evaluation of high-resolution melt curve analysis for rapid differentiation of Campylobacter hepaticus from other species in birds.
title_full_unstemmed	Evaluation of high-resolution melt curve analysis for rapid differentiation of Campylobacter hepaticus from other species in birds.
title_sort	evaluation of high-resolution melt curve analysis for rapid differentiation of campylobacter hepaticus from other species in birds.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/576e383e69744f5c9e5852f3972f8a44
work_keys_str_mv	AT petrinayoung evaluationofhighresolutionmeltcurveanalysisforrapiddifferentiationofcampylobacterhepaticusfromotherspeciesinbirds AT poltarce evaluationofhighresolutionmeltcurveanalysisforrapiddifferentiationofcampylobacterhepaticusfromotherspeciesinbirds AT sadhanaadhikary evaluationofhighresolutionmeltcurveanalysisforrapiddifferentiationofcampylobacterhepaticusfromotherspeciesinbirds AT joanneconnolly evaluationofhighresolutionmeltcurveanalysisforrapiddifferentiationofcampylobacterhepaticusfromotherspeciesinbirds AT timcrawshaw evaluationofhighresolutionmeltcurveanalysisforrapiddifferentiationofcampylobacterhepaticusfromotherspeciesinbirds AT seyedaghorashi evaluationofhighresolutionmeltcurveanalysisforrapiddifferentiationofcampylobacterhepaticusfromotherspeciesinbirds
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Evaluation of high-resolution melt curve analysis for rapid differentiation of Campylobacter hepaticus from other species in birds.

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