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首页> 外文期刊>RSC Advances >Identification of donkey meat in foods using species-specific PCR combined with lateral flow immunoassay
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Identification of donkey meat in foods using species-specific PCR combined with lateral flow immunoassay

机译：结合物种特异性PCR和侧向流免疫测定法鉴定食品中的驴肉

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Food authenticity is a global issue and has raised increasing concerns in the past decades. DNA-based methods are more favourable than the conventional protein-based techniques and have been applied to species identification and meat fraud detection. To effectively identify donkey meat for meat product authentication, a highly specific and robust method that coupled polymerase chain reaction (PCR) with lateral flow immunoassay (LFI) was developed. Donkey-specific PCR primers were designed by targeting at the mitochondrial D-loop gene and the specificity was verified in silico and in vitro against 22 species involved in meat authentication. A limit of detection of 0.0013 ng μL ~(?1) DNA extract was achieved and as low as 0.001% w/w (raw) and 0.01% w/w (cooked) donkey meat in beef were successfully detected using the developed PCR-LFI. LFI strip-based visualization of PCR products allowed for a 10-fold higher sensitivity than conventional gel electrophoresis and significantly reduced the analysis time for the post-PCR analysis. This PCR-LFI is highly suitable for rapid identification of donkey or incorporating into multiplex screening protocol for other meat authentication in the laboratories of both regulatory agencies and commercial services.
机译：食品的真实性是一个全球性问题，并且在过去的几十年中引起了越来越多的关注。基于DNA的方法比基于常规蛋白质的技术更有利，并且已应用于物种识别和肉类欺诈检测。为了有效地识别驴肉以进行肉制品认证，开发了一种高度特异性和鲁棒性的方法，该方法将聚合酶链反应（PCR）与侧向流免疫分析（LFI）结合在一起。通过靶向线粒体D-loop基因设计驴特异性PCR引物，并在计算机和体外针对涉及肉类鉴定的22种物种验证了特异性。使用开发的PCR-技术可成功检测到牛肉中0.0013 ngμL〜（？1）DNA提取物的检出限，并且成功检测出牛肉中低至0.001％w / w（生）和0.01％w / w（煮熟）的驴肉。 LFI。基于LFI条带的PCR产物可视化技术使灵敏度比常规凝胶电泳高10倍，并大大减少了PCR后分析的分析时间。该PCR-LFI非常适用于驴子的快速鉴定或在监管机构和商业服务机构的实验室中用于其他肉品鉴定的多重筛选方案中。

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