Detection of Escherichia coli and Salmonella from retail quail meat through optimized multiplex PCR

The conventional and multiplex pcr was developed for rapid and simultaneous detection of multiple food-borne pathogens in single reaction mixture. conventional pcr was optimized with annealing temperature at 50°c,each primer concentration of 10 pmol,dna template 200 ng and 10 μl fermentas master mix with 35 pcr cycles. similarly multiplex pcr was optimized with annealing temperature 56°c,each primer concentration of 20 pmol,dna template 200 ng using 23 μl qiagen multiplex master mix and 35 pcr cycles. a total 100 slaughtered quails from various chain stores (n=40),retail market (n=40) and university of veterinary and animal sciences quail farm (n=20) were processed for conventional and multiplex pcr for direct detection of escherichia coli and salmonella. prevalence of escherichia coli and salmonella through conventional culture method and biochemical testing was detected to be 82.5% and 66.6% respectively. while through conventional and multiplex pcr recovery of e. coli was 90% and salmonella was 82%. statistically no significant difference (p>0.05) was found between conventional culture method,conventional pcr and multiplex pcr. similarly no significant difference (p>0.05) was observed in recovery rate of escherichia coli and salmonella in quail meant collected from chain stores,retail market and uvas quail farm. © 2015 university of agriculture. all rights reserved.