Nanotechnology and Catalysis Research Centre (NANOCAT), University of Malaya, Kuala Lumpur, Malaysia.
Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2021 Aug;38(8):1273-1288. doi: 10.1080/19440049.2021.1925748. Epub 2021 Jun 2.
Food fraud is a global problem raising increased concerns during the past decades and food authenticity is now a burning issue. Beef, buffalo, chicken, duck, goat, sheep, and pork are heavily consumed meats bearing nutritional, economic and cultural/religious importance and are often found to be adulterated in raw and processed states. To authenticate these species, we developed and validated a highly specific multiplex (heptaplex) PCR assay targeting short length amplicons (73-263 bp) using seven pairs of species-specific primer sets targeting mitochondrial cytochrome b (cytb) and NADH dehydrogenase subunit 5 (ND5) genes. Specificity checking ( and ) against 25 non-target species revealed no cross-species amplification. The developed multiplex assay was validated with various adulterated and heat-treated (boiled, microwaved and autoclaved) meatball products and were found to show high sensitivity and stability under all processing conditions. The assay was sensitive enough to detect 0.01-0.005 ng of DNA from raw meat and 0.5% (w/w) adulterated meat in mixed matrices. A market survey revealed mislabelling of 95% beef and 15% chicken products while pork products were found pure. Given some advantageous features including short sizes of amplicons, exceptional stability and superior sensitivity, the developed assay could be conveniently used for discriminatory detection of target species with a variety of raw meat as well as processed meat products undergoing extreme processing treatments.
食品欺诈是一个全球性问题，在过去几十年中引起了越来越多的关注，食品真实性现在是一个热门话题。牛肉、水牛、鸡肉、鸭肉、羊肉、山羊肉和猪肉是人们大量消费的肉类，具有营养、经济和文化/宗教意义，而且经常在生肉和加工肉中发现被掺假。为了鉴定这些物种，我们开发并验证了一种针对短长度扩增子（73-263 bp）的高度特异性多重（七重）PCR 检测方法，使用七对针对线粒体细胞色素 b（cytb）和 NADH 脱氢酶亚基 5（ND5）基因的种特异性引物对。特异性检查（和）对 25 种非目标物种没有发现交叉物种扩增。开发的多重检测方法已用于各种掺假和热处理（煮沸、微波和高压灭菌）的肉丸产品，并在所有加工条件下均显示出高灵敏度和稳定性。该检测方法足够灵敏，可从生肉中检测到 0.01-0.005 ng 的 DNA，在混合基质中可检测到 0.5%（w/w）掺假肉。市场调查显示，95%的牛肉和 15%的鸡肉产品存在标签错误，而猪肉产品则是纯的。鉴于一些有利的特点，包括扩增子的短尺寸、出色的稳定性和卓越的灵敏度，开发的检测方法可方便地用于鉴别检测目标物种的各种生肉和经过极端加工处理的加工肉产品。
