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

Abstract

The adulteraon of processed beef-based meat products with pork is a sensive issue in Indonesia. Therefore a simple, low cost, and accurate method is required for the detecon of pork, so as to protect consumers from accidental consumpon of adulterated meat. In this study, we developed a detecon method for the low cost idenficaon 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 detecon. A posive control containing a pork gene inserted into pGEM®-T easy was prepared, along with a negave control. The results of the duplex PCR were used to assess its specificity, detecon limit, and its ability to recognize pork in processed meat products with a different food matrix. 18S rRNA detecon was for confirming DNA integrity of DNA extracted from the processed food, while the posive control confirmed that the reagents were working well and the negave control confirmed a non-contaminaon problem. Following this, the duplex PCR was opmized and the opmum concentraon primer for duplex PCR detecon was found to be 3 µM for pork and 0.2 µM for 18S rRNA. As lile 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, sensive, specific, and low cost method of detecng pork in processed meat products.