|
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
Department of Microbiology, Faculty of Specialized
Veterinary Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.
*Corresponding author. E-mail:
tsalehi@ut.ac.ir.
Accepted 22 July, 2010 |
|
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.
Key words:
Salmonella, poultry, polymerase chain reaction
restriction fragment length polymorphism, groEL. |
