Differentiation of salmonella isolates using PCR and high-resolution melt curve analysis

Seyed Ghorashi, Mohammad Saeidabadi, Hassan Nili, Habibollah Dadras, Hassan Sharifiyazdi, Joanne Connolly, Mary Valcanis, Shane Raidal

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Abstract

Poultry products can be contaminated with Salmonella isolates which may cause foodborne disease in humans. The aim of this study was to optimise a rapid diagnostic test that can differentiate Salmonella isolates without requiring DNA sequencing. Specific primers were used to amplify hem-D gene of 16 different Salmonella reference strains and 65 clinical isolates from commercial poultry farms. PCR products were applied to high-resolution melt curve analysis and Salmonella isolates were differentiated based on their HRM curves. Analysis of the nucleotide sequence of the amplicons from selected isolates confirmed that each melting curve profile was related to a unique DNA sequence. The relationship between reference strains and tested specimens was also evaluated using a mathematical model without visual interpretation of HRM curves. In addition, the potential of the PCR-HRM curve analysis was evaluated for genotyping of additional Salmonella isolates from different avian species. The findings indicate that PCR followed by HRM curve analysis provides a rapid and robust technique for genotyping of Salmonella isolates to determine the Salmonella strains.

Key words: Salmonella, PCR, high resolution melting curve analysis, strain differentiation

Original languageEnglish
Pages33
Number of pages1
Publication statusPublished - 05 Oct 2017
Event2nd International Congress on Advances in Veterinary Sciences & Technics: ICAVST - Skopje, Macedonia, The Former Yugoslav Republic of
Duration: 04 Oct 201708 Oct 2017
https://www.icavst.com/icavst-2017

Conference

Conference2nd International Congress on Advances in Veterinary Sciences & Technics
CountryMacedonia, The Former Yugoslav Republic of
CitySkopje
Period04/10/1708/10/17
Internet address

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