Direct PCR-Based Detection of Pork Adulteration in Food Products Using ND4 and Cytb Markers

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Joni Kusnadi, Nur Laily Fera Ari Sutejo

2026 Pertanika Journal of Tropical Agricultural Science Vol. 49 Issue 1 Article Cited by 0

Abstract

The adulteration of edible processed meat has become widespread, as low-quality meat is added. One of the most prevalent instances is the use of an adulterated meat product containing pork, which caused problems for Muslims because of their dietary restrictions. Food safety needs a quick, easy, and specific DNA-based detection method. Therefore, the present study sought to detect pork contamination utilising the direct-PCR method with NADH dehydrogenase 4 (ND4) and cytochrome b (Cyt-b) primers in fresh pork and commercially processed meat products. The lysis buffer recipe was optimised using two temperatures and three incubation periods. Conventional PCR and electrophoresis were used to confirm DNA detection. Three brands of meatballs, sausages, and corned beef obtained from several markets in Malang City, Indonesia, were examined following optimisation. With high levels of DNA concentration and purity as well as visible DNA bands, the incubation temperature of 95°C for 5 minutes clearly exhibited the best results. All ND4 and Cyt-b primers amplified pork DNA in the majority of the processed samples, except for some Cyt-b primers, which did not amplify DNA in certain corned beef products. The direct-PCR technique is clearly a straightforward, fast, inexpensive, and reliable method for the detection of pork adulteration in processed meat products and could also be applied for halal food authentication. © Universiti Putra Malaysia Press.

Affiliations

Department of Agriculture Product Technology, Faculty of Agriculture Technology, Universitas Brawijaya, East Java, Malang, 65145, Indonesia