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

IF 4.7 Q2 FOOD SCIENCE & TECHNOLOGY

Food Chemistry Molecular Sciences Pub Date : 2023-08-12 DOI:10.1016/j.fochms.2023.100181

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

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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.

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验证针对ND2的双链PCR在肉制品中检测牛和猪

食品认证是确保公平贸易的强制性措施。本研究从NADH脱氢酶亚基2（ND2）基因开发了一种双链聚合酶链式反应（PCR）来扩增牛和猪DNA的特定片段。用一个由19个碱基对（bp）组成的通用正向引物（3′-CAAACAAACTCACTCGAAAA-5′）和一个由20个（3′-CCAAACACACTCGAAA-A-5′）和21个（3’-TGGCAAGAATTAGGAGGTTA-5’）bp组成的物种特异性反向引物来限制猪和牛的扩增DNA片段。PCR反应将产生分别具有168和227bp轮廓的产物。为了研究检测的准确性和限度，对从印度尼西亚苏拉卡塔的商业市场随机购买的商业肉丸进行了一项使用单纯形和双链PCR的体外实验。本研究结果表明，ND2可作为肉制品中猪和牛肉品种鉴定的替代遗传标记。

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来源期刊

Food Chemistry Molecular Sciences Agricultural and Biological Sciences-Food Science

CiteScore

6.00

自引率

0.00%

发文量

审稿时长

82 days

期刊介绍： Food Chemistry: Molecular Sciences is one of three companion journals to the highly respected Food Chemistry. Food Chemistry: Molecular Sciences is an open access journal publishing research advancing the theory and practice of molecular sciences of foods. The types of articles considered are original research articles, analytical methods, comprehensive reviews and commentaries. Topics include: Molecular sciences relating to major and minor components of food (nutrients and bioactives) and their physiological, sensory, flavour, and microbiological aspects; data must be sufficient to demonstrate relevance to foods and as consumed by humans Changes in molecular composition or structure in foods occurring or induced during growth, distribution and processing (industrial or domestic) or as a result of human metabolism Quality, safety, authenticity and traceability of foods and packaging materials Valorisation of food waste arising from processing and exploitation of by-products Molecular sciences of additives, contaminants including agro-chemicals, together with their metabolism, food fate and benefit: risk to human health Novel analytical and computational (bioinformatics) methods related to foods as consumed, nutrients and bioactives, sensory, metabolic fate, and origins of foods. Articles must be concerned with new or novel methods or novel uses and must be applied to real-world samples to demonstrate robustness. Those dealing with significant improvements to existing methods or foods and commodities from different regions, and re-use of existing data will be considered, provided authors can establish sufficient originality.