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

Authors

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

DOI:

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

Keywords:

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

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.

Author Biographies

Joni Kusnadi, University of Brawijaya, Central Laboratory of Life Sciences & Halal Inspection Agency University of Brawijaya

Khotibul Umam Al-Awwaly, University of Brawijaya & Halal Inspection Agency University of Brawijaya

Estri Laras Arumingtyas, University of Brawijaya

Nur Istianah, Kyungpook National University

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Published

2022-08-31

How to Cite

Kusnadi, J., Hernandi, K. H., Al-Awwaly, K. U., Arumingtyas, E. L., Hakiki, H. M., & Istianah, N. (2022). Design and Performance Test of Specific Primers to Detect Bovine DNA Fragments using Multiplex PCR Technique for Halal Authentification. Indonesian Journal of Halal Research, 4(2), 45–52. https://doi.org/10.15575/ijhar.v4i2.15573

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