Validating duplex-PCR targeting ND2 for bovine and porcine detection in meat products

Farouq Heidar Barido; Desti Desti; Ahmad Pramono; Zakaria Husein Abdurrahman; Slamet Diah Volkandari; Muhammad Cahyadi

doi:10.1016/j.fochms.2023.100181

Validating duplex-PCR targeting ND2 for bovine and porcine detection in meat products

Food Chem (Oxf). 2023 Aug 12:7:100181. doi: 10.1016/j.fochms.2023.100181. eCollection 2023 Dec 30.

Authors

Farouq Heidar Barido^{1

2}, Desti Desti¹, Ahmad Pramono¹, Zakaria Husein Abdurrahman³, Slamet Diah Volkandari⁴, Muhammad Cahyadi^{1

2}

Affiliations

¹ Faculty of Animal Science, Universitas Sebelas Maret, Surakarta, Jawa Tengah 57126, Indonesia.
² Halal Research Center and Services (HRCS), Institute for Research and Community Service, Universitas Sebelas Maret, Surakarta, Jawa Tengah 57126, Indonesia.
³ Faculty of Animal Science, Universitas Boyolali, Boyolali, Jawa Tengah 57315, Indonesia.
⁴ Research Center for Food Technology and Processing (PRTPP), National Research and Innovation Agency (BRIN), Gunung Kidul, Daerah Istimewa, Yogyakarta 55861, Indonesia.

Abstract

Food authentication is a mandatory effort to assure the fair-trade. This study developed a duplex polymerase chain reaction (PCR) from the NADH dehydrogenase subunit 2 (ND2) gene to amplify specific segments of a cattle and porcine DNA. A universal forward primer composed of nineteen base pairs (bp) (3'-CCAAACACAACTCCGAAAA-5') and species-specific reverse primers composed of twenty (3'-CCAAACACAACTCCGAAAA-5') and twenty-one (3'-TGGCAAGAATTAGGACGGTTA-5') bp were used to limit the amplified DNA segment for porcine and cattle. The PCR reaction would generate a product with a profile of 168 and 227 bp, respectively. To investigate the accuracy and limit of detection, an in vitro experiment was conducted using simplex and duplex PCR on commercial meatballs randomly purchased from a commercial market in Surakarta, Indonesia. The findings of this study indicated that ND2 could be used as an alternative genetic marker for the identification of porcine and beef species in meat-derived products.

Keywords: PCR; Primer; Primer design; Species identification; mt-DNA ND2.