For revealing animal species in complex or adulterated processed meat product, we presented a method combining a novel cytochrome oxidase I (COI) mini-barcode with next-generation sequencing (NGS), which identifies various animal species (swine, bovine, Caprinae, and some of fish, shrimp, and poultry) accurately and efficiently in processed meat products. We designed a universal primer based on 140 sequences from 51 edible animal species. A mixture of 12 species raw meat samples were identified with the clone sequencing and also with a mini-barcode- (136 bp) combined NGS method, respectively. The mini-barcode of these 12 species was 100% identical to the target species sequence by Sanger sequencing. Compared to the clone sequencing method, the NGS method is superior in accuracy, sensitivity, and detection efficiency. Various edible animal species were identified in the species level both in the mixed samples and the 7 heavily processed food products. Moreover, some unlabeled species and dubious contamination were detected as well.

中文翻译：

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将 COI 迷你条形码与下一代测序相结合，用于加工肉类产品中的动物源成分识别

为了揭示复杂或掺假加工肉制品中的动物物种，我们提出了一种将新型细胞色素氧化酶 I (COI) 迷你条形码与下一代测序 (NGS) 相结合的方法，该方法可识别各种动物物种（猪、牛、山羊和一些鱼、虾和家禽）在加工肉制品中准确有效。我们基于 51 个可食用动物物种的 140 个序列设计了一个通用引物。分别使用克隆测序和迷你条形码（136 bp）组合 NGS 方法鉴定了 12 种生肉样品的混合物。通过 Sanger 测序，这 12 个物种的迷你条形码与目标物种序列 100% 相同。与克隆测序方法相比，NGS方法在准确度、灵敏度和检测效率方面具有优越性。在混合样本和 7 种高度加工食品中，在物种水平上确定了各种可食用动物物种。此外，还检测到一些未标记的物种和可疑的污染。