Design and Performance Test of Specific Primers to Detect Bovine DNA Fragments using Multiplex PCR Technique for Halal Authentification

Joni Kusnadi (University of Brawijaya, Central Laboratory of Life Sciences & Halal Inspection Agency University of Brawijaya)
Kevin Hohn Hernandi (University of Brawijaya)
Khotibul Umam Al-Awwaly (University of Brawijaya & Halal Inspection Agency University of Brawijaya)
Estri Laras Arumingtyas (University of Brawijaya)
Hilda Maghfirotu Hakiki (Halal Inspection Agency University of Brawijaya)
Nur Istianah (Kyungpook National University)

Indonesian Journal of Halal Research
Vol 4, No 2 (2022): August

Adulterating meat products with several species, including non-halal species, is often found in commercial products. This study aims to design and validate the Cytochrome c oxidase subunit I (CO1) primers to detect the non-halal species. A pair of species-specific primers encoding the CO1 gene were designed to amplify bovine DNA, tested for specificity, and applied in multiplex polymerase chain reaction (PCR) technique with D-loop primers for pigs, Cyt-b for rats, and 12S rRNA for dogs. The CO1 primers, along with D-loop primers for porcine, Cyt-b primers for rats, and 12S rRNA primers for dogs can be used to detect specific bovine DNA with a size of 279 bp and sequence similarity of 96%. In addition, dog, rat, and porcine DNA were amplified at 101, 603, and 951 bp, respectively. These four primers are specific and can amplify the target DNA to detect non-halal meat component contamination in a single reaction process.