Detection of Rusa spp. DNA in ready-to-eat food (RTF) using singleplex of Polymerase Chain Reaction

Abstract

Deer meat is more expensive than meat from domesticated animals, making it a possible target for adulteration. As a result, a practical approach for detecting deer DNA was developed by employing a Polymerase Chain Reaction (PCR) assay to target a region of the mitochondrial Cytochrome Oxidase Subunit I (COI) gene in locally ready-to-eat food (RTE) products. A specificity trial was performed on eight deer-based RTF to include Kerutup Rusa, Kari Tulang Rusa, Kurma Rusa, and Rendang Tok Rusa food matrices (to include the four commercially processed RTE food products) and deer DNA was detected from four commercial RTE products (Kerutup Rusa, Kari Tulang Rusa, Kurma Rusa and Rendang Tok Rusa) with the CerV primers at 0.0001 ng/μL detection limit, showing that both the primers and the assays were effective at detecting DNA targets in thermally treated matrices. This assay addresses critical gaps in halal certification and food fraud prevention, offering a reliable tool for regulatory enforcement and consumer protection in Malaysia's growing RTE market. The CerV gene detection limit was 0.0001 ng of CRM, consistent with the Certified Reference Material (CRM) used in this investigation (Red Deer). As a result of the present investigation, it is clear that PCR targeting the CerV COI 1 gene is appropriate for identifying deer DNA in RTE food samples.