Low cost and comprehensive pork detection in processed food products with a different food matrix

Abstract

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.