OPTIMIZATION OF MULTIPLEX PCR FOR DETECTION OF GRAM POSITIVE FOODBORNE PATHOGENS FROM RETAIL QUAIL MEAT

This experimental design was developed to optimize the conventional and multiplex PCR reaction for detection of Staphylococcus aureus and Bacillus cereus from quail meat samples. Conventional PCR was optimized with annealing temperature 50 degrees C, each primer concentrations of 10pmol, 200ng of DNA template and 10 mu l Qiagen master mix and 35 number of PCR cycles. In the same way multiplex PCR was optimized with annealing temperature of 50 degrees C, 20pmol of each primer, 200ng DNA template of each bacteria and 23 mu l of Qiagen multiplex master mix. Quail meat samples were collected from chain stores (n=40), retail market (n=40) and UVAS quail farm (n=20). Prevalence of S. aureus through conventional culture method was detected to be 80 percent. While through conventional and multiplex PCR recovery was 85 percent. Prevalence of B. cereus through conventional culture method was 65 percent, while through conventional and multiplex PCR recovery rate was 72 percent. 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 S. aureus and B. cereus in quail meat collected from chain stores, retail market and UVAS quail farm.