基于微型DNA条形码的多种动物源性成分的鉴定

Mini-DNA Barcode for Identification of Multiple Animal-Derived Ingredients

通过分析51种动物的DNA条形码序列,新设计一对微型DNA条形码的通用引物,建立一种联合微型DNA条形码和克隆测序法对动物源性多组分样品进行准确、高效鉴定的方法。用微型DNA条形码分别鉴定11种常见食用肉类,判断其对物种鉴定的准确性,并比较其与标准DNA条形码对物种的鉴定效果,然后将11种常见商品肉类等比例混合,探究克隆测序法对混合肉样品的鉴定效果。结果表明:引物有较好的通用性,能对具有不同的引物同源性水平的11个物种进行有效扩增并获单一条带。微型条形码能准确鉴定11个肉类物种,与常规DNA条形码鉴定效果一致。经克隆测序法,把混合样品的9个物种一并检出,总体上达到了较高的检出效率,同时深入讨论了未能检出全部物种的原因。微型条形码良好的物种鉴定能力的研究结果为联合微型DNA条形码与二代测序技术进行混合样品中物种的高效鉴定提供参考依据。

Based on the standard DNA barcodes of 51 animal species, a new pair of universal primers for mini-DNA barcode was designed and combined with clone sequencing to develop an accurate and efficient method for identifying multiple animal derived ingredients. Eleven common edible meat species were identified by the mini-DNA barcode to verify its accuracy in comparison with the results of standard DNA barcode. A mixture in equal proportion of the 11 species was used to evaluate the colon sequencing method. The results showed that the designed primers had favorable universality among 11 species with different levels of sequence homology with primers. Thus, 11 meat species could be accurately identified with the mini-barcode, which showed high consistency to the standard DNA barcode. By clone sequencing, nine species from mixed samples were detected. Furthermore, the reasons why not all the species were detected were discussed. The results of this study can lay a foundation for combining mini-DNA barcode and next generation sequencing technique for efficient species identification in mixed meat.