Identification of Salmonella isolated from poultry by MPCR technique and evaluation of their hsp groEL gene diversity based on the PCR-RFLP analysis


J. Akbarmehr, T. Zahraei Salehi* and G. H. Nikbakht Brujeni

The aim of this study was to isolate Salmonella from poultry and evaluation of their hsp groEL gene diversity by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis. In this research 58 strains of 3 different Salmonella serogroups (D1, B and C) were isolated from poultry farms of East Azarbayjan province of Iran by bacteriological and biochemical tests. For confirmation of Salmonella typhimurium and Salmonella enteritidis serovars multiplex polymerase chain reaction (PCR) was applied with four pairs of primers for S. typhimurium and three pairs of primers for Salmonella Enteritidis. PCR-RFLP analysis was carried out on the 1.6 kb groEL gene for evaluation of their hsp groEL gene diversity. The data generated by multiplex polymerase chain reaction (MPCR) method indicated that strains of S. enteritidis (serogroup D1) and S. typhimurium (serogroup B) were the most common isolates. Amplification of the groEL gene produced an identical profile for all the 58 Salmonella strains. Hae III restriction enzyme was used to restrict the groEL gene for PCR-RFLP analysis. Based on the results of this experiment digested groEL gene of the S. typhimurium strains produced four Hae III restricted bands between 150 - 850 bp and serovars belonging to S. enteritidis strains produced five Hae III restricted bands between 150 - 630 bp. Strains belonging to serogroup C produced a combination of five and four restricted bands similar to S. enteritidis and S. typhimurium respectively. This study showed that there were differences in the Hae III restriction sites within the groEL gene of strains belonging to serovars S. typhimurium and S. enteritidis but clear discrimination between the serovars of different Salmonella serogroups was not observed.

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