Razzak Md Abdur, Hamid Sharifah Bee Abd, Ali Md Eaqub
a Nanotechnology and Catalysis Research Centre (NanoCat) , University of Malaya , Kuala Lumpur 50603 , Malaysia.
b Centre for Research in Biotechnology for Agriculture (CEBAR) , University of Malaya , Kuala Lumpur 50603 , Malaysia.
Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2015;32(11):1902-13. doi: 10.1080/19440049.2015.1087060. Epub 2015 Oct 8.
Food forgery has posed considerable risk to public health, religious rituals, personal budget and wildlife. Pig, dog, cat, rat and monkey meat are restricted in most religions, but their sporadic adulteration are rampant. Market controllers need a low-cost but reliable technique to track and trace suspected species in the food chain. Considering the need, here we documented a lab-on-a-chip-based multiplex polymerase chain reaction (PCR) assay for the authentication of five non-halal meat species in foods. Using species-specific primers, 172, 163, 141, 129 and 108-bp sites of mitochondrial ND5, ATPase 6 and cytochrome b genes were amplified to detect cat, dog, pig, monkey and rat species under complex matrices. Species-specificity was authenticated against 20 different species with the potential to be used in food. The targets were stable under extreme sterilisation (121°C at 45 psi for 2.5 h) which severely degrades DNA. The assay was optimised under the backgrounds of various commercial meat products and validated for the analysis of meatballs, burgers and frankfurters, which are popular fast food items across the globe. The assay was tested to detect 0.1% suspected meats under commercial backgrounds of marketed foods. Instead of simplex PCR which detects only one species at a time, such a multiplex platform can reduce cost by at least fivefolds by detecting five different species in a single assay platform.
食品造假对公众健康、宗教仪式、个人预算和野生动物构成了相当大的风险。猪肉、狗肉、猫肉、鼠肉和猴肉在大多数宗教中都是受限的，但它们的零星掺假现象却很猖獗。市场监管者需要一种低成本但可靠的技术来追踪和溯源食物链中可疑的物种。考虑到这一需求，我们在此记录了一种基于芯片实验室的多重聚合酶链反应（PCR）检测方法，用于鉴定食品中的五种非清真肉类物种。使用物种特异性引物，扩增线粒体ND5、ATPase 6和细胞色素b基因的172、163、141、129和108碱基对位点，以检测复杂基质下的猫、狗、猪、猴和鼠物种。针对20种可能用于食品的不同物种验证了物种特异性。目标在极端灭菌条件下（121°C，45磅力/平方英寸，2.5小时）保持稳定，这种条件会严重降解DNA。该检测方法在各种商业肉类产品背景下进行了优化，并针对全球流行的快餐食品肉丸、汉堡和法兰克福香肠的分析进行了验证。该检测方法经过测试，可在市售食品的商业背景下检测出0.1%的可疑肉类。与一次仅检测一个物种的单重PCR不同，这种多重平台通过在单个检测平台上检测五种不同物种，可将成本降低至少五倍。
