Evaluation of PCR and high resolution melt curve analysis for differentiation of Salmonella isolates

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

Research output: Contribution to journalArticlepeer-review

4 Citations (Scopus)

Abstract

Consumption of poultry products contaminated with Salmonella is one of the major causes of foodborne diseases worldwide and therefore detection and differentiation of Salmonella spp. in poultry is important. In this study, oligonucleotide primers were designed from hemD gene and a PCR followed by high-resolution melt (HRM) curve analysis was developed for rapid differentiation of Salmonella isolates. Amplicons of 228 bp were generated from 16 different Salmonella reference strains and from 65 clinical field isolates mainly from poultry farms. HRM curve analysis of the amplicons differentiated Salmonella isolates and analysis of the nucleotide sequence of the amplicons from selected isolates revealed 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 serovar/serotype.

Original languageEnglish
Pages (from-to)319-331
Number of pages27
JournalAvian Pathology
Volume46
Issue number3
DOIs
Publication statusPublished - 2017

Fingerprint

Dive into the research topics of 'Evaluation of PCR and high resolution melt curve analysis for differentiation of Salmonella isolates'. Together they form a unique fingerprint.

Cite this