title: Evaluation of PCR and high-resolution melt curve analysis for differentiation of Salmonella isolates.
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.
authors.affiliation: a School of Veterinary Medicine , Shiraz University , Shiraz , Iran.|b School of Animal and Veterinary Sciences , Charles Sturt University , Wagga Wagga , Australia.
authors.forename: Mohammad Sadegh
authors.initials: MS
authors.lastname: Saeidabadi
authors.affiliation: a School of Veterinary Medicine , Shiraz University , Shiraz , Iran.
authors.forename: Hassan
authors.initials: H
authors.lastname: Nili
authors.affiliation: a School of Veterinary Medicine , Shiraz University , Shiraz , Iran.
authors.forename: Habibollah
authors.initials: H
authors.lastname: Dadras
authors.affiliation: a School of Veterinary Medicine , Shiraz University , Shiraz , Iran.
authors.forename: Hassan
authors.initials: H
authors.lastname: Sharifiyazdi
authors.affiliation: b School of Animal and Veterinary Sciences , Charles Sturt University , Wagga Wagga , Australia.|c Graham Centre for Agricultural Innovation (NSW Department of Primary Industries and Charles Sturt University) , Wagga Wagga , Australia.
authors.forename: Joanne
authors.initials: J
authors.lastname: Connolly
authors.affiliation: d Department of Microbiology and Immunology , The University of Melbourne , Parkville , Australia.
authors.forename: Mary
authors.initials: M
authors.lastname: Valcanis
authors.affiliation: b School of Animal and Veterinary Sciences , Charles Sturt University , Wagga Wagga , Australia.|c Graham Centre for Agricultural Innovation (NSW Department of Primary Industries and Charles Sturt University) , Wagga Wagga , Australia.
authors.forename: Shane
authors.initials: S
authors.lastname: Raidal
authors.affiliation: b School of Animal and Veterinary Sciences , Charles Sturt University , Wagga Wagga , Australia.|c Graham Centre for Agricultural Innovation (NSW Department of Primary Industries and Charles Sturt University) , Wagga Wagga , Australia.
authors.forename: Seyed Ali
authors.initials: SA
authors.lastname: Ghorashi
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques / Q000662:veterinary; D017931:DNA Primers / Q000235:genetics; D005838:Genotype; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011200:Poultry; D011201:Poultry Diseases / Q000382:microbiology*; D012475:Salmonella / Q000235:genetics / Q000302:isolation & purification*; D012481:Salmonella Infections, Animal / Q000382:microbiology*; D013045:Species Specificity; D044366:Transition Temperature
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers
keywords: PCR; Salmonella; genotyping; high-resolution melting curve analysis; strain differentiation