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


Joni Kusnadi(1*), Kevin Hohn Hernandi(2), Khotibul Umam Al-Awwaly(3), Estri Laras Arumingtyas(4), Hilda Maghfirotu Hakiki(5), Nur Istianah(6)

(1) University of Brawijaya, Central Laboratory of Life Sciences & Halal Inspection Agency University of Brawijaya, Indonesia
(2) University of Brawijaya, Indonesia
(3) University of Brawijaya & Halal Inspection Agency University of Brawijaya, Indonesia
(4) University of Brawijaya, Indonesia
(5) Halal Inspection Agency University of Brawijaya, Indonesia
(6) Kyungpook National University, Korea, Republic of
(*) Corresponding Author

Abstract


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.


Keywords


bovine; CO1 primer; halal authentification; meat products; multiplex PCR

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References


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DOI: https://doi.org/10.15575/ijhar.v4i2.15573

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