The adultera on of processed beef-based meat products with pork is a sensi ve issue in Indonesia. Therefore a simple, low cost, and accurate method is required for the detec on of pork, so as to protect consumers from accidental consump on of adulterated meat. In this study, we developed a detec on method for the low cost iden fica on of pork in processed meat products. We used the cost-efficient Taq DNA polymerase, DreamTaq Green PCR master mix (2x), and duplex PCR method to recognize pork simultaneously with 18S rRNA detec on. A posi ve control containing a pork gene inserted into pGEM ® -T easy was prepared, along with a nega ve control. The results of the duplex PCR were used to assess its specificity, detec on limit, and its ability to recognize pork in processed meat products with a different food matrix. 18S rRNA detec on was for confirming DNA integrity of DNA extracted from the processed food, while the posi ve control confirmed that the reagents were working well and the nega ve control confirmed a non-contamina on problem. Following this, the duplex PCR was op mized and the op mum concentra on primer for duplex PCR detec on was found to be 3 µM for pork and 0.2 µM for 18S rRNA. As li le as 3.125 ng of the DNA template could be used to detect whether a sample contained pork. Of the nine commercial processed meat products tested, five were found to contain pork while four halal products showed no signs of pork. It can be concluded that duplex PCR is a simple, fast, sensi ve, specific, and low cost method of detec ng pork in processed meat products.

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.