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Pork contamination in food and food products is a major concern affecting consumer confidence in halal foods. This work aims to develop a DNA marker for detecting contaminated porcine DNA in halal food and food products using the Polymerase Chain Reaction (PCR) method to amplify growth hormone (GH) intron, 105 base pairs, in which other standardized methods are limited in their sensitivities and long testing process. When validating the developed DNA marker, 4 meat sources: beef (Bos taurus), pork (Sus scrofa domesticus), wild boar (Sus scrofa), and chicken (Gallus gallus), 1 plant starch: maize (Zea mays), and 5 pork-based products: fragrance pork floss, Vietnamese sausage, ham, pork sausage, and Chinese sausage were examined. The PCR results showed positive given a 105-bp DNA band on pork, wild boar, and all pork-based products, but no amplified products in other tested samples except for beef and mixed beef samples, where two off-target DNA bands were present at approximately 270 and 1,100 bp. The PCR sensitivity was additionally conducted using beef-pork binary admixtures in different pork percentages (w/w) at 50, 25, 5, 1, and 0.1 (25, 12.5, 2.5, 0.5, and 0.05 ng, respectively). The results showed high sensitivity, detecting as low as 0.1% (0.05 ng). The PCR sensitivity was also performed using a mixture of bovine and porcine DNAs in various percentages (v/v) of porcine DNA: 50, 25, 5, 1, 0.1, and 0.01 (25, 12.5, 2.5, 0.5, 0.05, and 0.005 ng, respectively). The results enabled the detection of as little as 0.01% (0.005 ng) of porcine DNA. The pork derivatives in 5 halal foods were also investigated. The PCR gave positive results in 2 halal foods. Thus, the DNA marker and PCR method developed in this study are highly sensitive, easy to use, and can apply to detecting porcine DNA presence in halal foods and food products.
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