Triplex PCR for halal authentication of processed food: Development and Characterization

Abstract

Awareness of consumers towards the safety of food and labelling statement due to food adulterations is increasing. Simultaneous detection made the developed methods are rapid and economic. In this study we developed triplex PCR for beef and pork detection by using 18SsRNA as the internal control, then further characterized the methods. Primer formulation for triplex PCR being used are 0, 8µM porcine, 0, 04 µM for each beef and 18sRNA, that all together working at 45°C annealing temperature in one single tube. The amplicon sizes for pork, beef and 18SrRNA are 300, 120 and 99bp respectively. The sensitivity of the method is 0, 851ng. This developed method shown as robust method that can detect DNA target from different source of matrices [five products contain pork (meatball, sausage, ham, pasta, cornet), 3 beef processed food (dendeng, rendang and satay) and three products contain pork include beef (sauce curry from three different products)] which could contain some types of PCR inhibitor. Furthermore, the method shows specific detection towards the target that had no cross contamination.