PCR-RFLP Analysis of a fliC Gene Fragment in Avian Salmonella Isolates

Salmonella are a genus of zoonotic bacteria of worldwide economic and health importance. Members of Salmonella enterica subspecies enterica are mainly associated with warm-blooded vertebrates and are usually transmitted by ingestion of food or water contaminated by infected feces. The aim of this study was to apply a PCR-RFLP method based on the fliC gene to identify the serotypes of Salmonella isolates from Karaj, Iran. A total of 30 Salmonella isolates were serotyped by specific antisera. For the PCR-RFLP method based on the fliC gene, extracted DNA was used as the template for amplifying the fliC gene (1500bp) using specific primers. PCR products were subjected to digestion using HhaI restriction endonuclease. Results: This study determined 30 serotypes as Salmonella durban (56.6%), Salmonella uno (23.3%), Salmonella enteritidis (3.3%), Salmonella tinda (3.3%), Salmonella mjimweme (3.3%), Salmonella Thompson (3.3%), Salmonella sIIO8 (3.3 %) and Salmonella sIIO7 (3.3%). Observations indicated that HhaI is able to discriminate Salmonella tinda and Salmonella thompson , yet Salmonella enteritidis , Salmonella durban and Salmonella mjimweme had the same pattern with this enzyme. Also Salmonella sIIO8 , Salmonella sIIO7 and Salmonella uno showed the same pattern. Thus, regarding the size and the number of resulting fragments from this enzyme, four patterns were obtained for HhaI. A large number of Salmonella serotypes need to be analyzed by the PCR-RFLP method and different enzymes must be used to give reliable results.

Salmonella are a genus of zoonotic bacteria of worldwide economic and health importance. Members of Salmonella enterica subspecies enterica are mainly associated with warm-blooded vertebrates and are usually transmitted by ingestion of food or water contaminated by infected feces. The aim of this study was to apply a PCR-RFLP method based on the fliC gene to identify the serotypes of Salmonella isolates from Karaj, Iran. A total of 30 Salmonella isolates were serotyped by specific antisera. For the PCR-RFLP method based on the fliC gene, extracted DNA was used as the template for amplifying the fliC gene (1500bp) using specific primers. PCR products were subjected to digestion using HhaI restriction endonuclease. Results: This study determined 30 serotypes as Salmonella durban (56.6%), Salmonella uno (23.3%), Salmonella enteritidis (3.3%), Salmonella tinda (3.3%), Salmonella mjimweme (3.3%), Salmonella Thompson (3.3%), Salmonella sIIO8 (3.3 %) and Salmonella sIIO7 (3.3%). Observations indicated that HhaI is able to discriminate Salmonella tinda and Salmonella thompson , yet Salmonella enteritidis , Salmonella durban and Salmonella mjimweme had the same pattern with this enzyme. Also Salmonella sIIO8 , Salmonella sIIO7 and Salmonella uno showed the same pattern. Thus, regarding the size and the number of resulting fragments from this enzyme, four patterns were obtained for HhaI. A large number of Salmonella serotypes need to be analyzed by the PCR-RFLP method and different enzymes must be used to give reliable results.