Application of direct PCR technique using specified mitochondrial DNAencoded ND4 and D-Loop genes on detecting pork DNA in processed meat products

Abstract

Adulteration or counterfeiting of food products raises public concerns about the halal issue. Many issues related to food products such as meatballs, sausages, and corned beef use pork for counterfeiting. This study aimed to determine a direct polymerase chain reaction (dPCR) technique for pork DNA detection in processed meat products such as meatballs, sausages, and corned beef in a short time and at a low cost using specific primers, ND4 and D-Loop. The study used a direct PCR technique with variations in the incubation time and temperature of the lysis process on the sample. The concentration of DNA obtained from the lysis results with variations in incubation time and temperature in fresh meat and processed meat products was 66.17-469.23 ng/µL, and the purity of DNA in fresh and processed meat products was 1.70-2.25. ND4 primer was more sensitive to DNA amplification than D-Loop in processed meat products using direct PCR. Meatballs, sausages, and corned beef products written on the packaging using beef did not detect any mixing or addition of pork. Direct PCR can detect pork DNA in processed meat products such as meatballs, sausages and corned beef.