The continuous development of fast and simple new methods to identify animal-derived ingredients is very important for the authentication of meat products. This study intended to develop a multiplex PCR method using new species-specific nuclear DNA (nDNA) sequences for the detection of ingredients derived from sheep/goat, bovine, chicken, duck and pig in meat products. Sequence alignment analysis in 53 species showed high specificity of species-specific nDNA. Species-specific primers were designed on the conservative region of each species-specific nDNA sequence. The specificity and conservation of the sequences and primers were verified by PCR reaction and sequencing with the limit of detection down to 0.5 ng. Then, a species-specific multiplex PCR method was developed and optimized to simultaneously detect sheep/goat (237 bp), bovine (223 bp), chicken (192 bp), duck (168 bp) and pig (154 bp) in one reaction. Various processed meat products containing one or more animal-derived ingredients were detected by the developed multiplex PCR method, and the results were consistent with their labeled meat species. Our study provides a fast and simple detection method for regulating labeling of animal-derived ingredients in meat products.

中文翻译：

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使用新颖的物种特异性核DNA序列检测肉制品中五种动物的多重PCR方法

识别动物源成分的快速简便的新方法的不断发展对于肉制品的认证非常重要。这项研究旨在开发一种使用新的物种特异性核DNA（nDNA）序列的多重PCR方法，以检测肉类产品中绵羊/山羊，牛，鸡，鸭和猪的成分。在53个物种中进行的序列比对分析表明，物种特异性nDNA具有高度特异性。在每个物种特异性nDNA序列的保守区域设计物种特异性引物。通过PCR反应和测序验证序列和引物的特异性和保守性，检测限低至0.5 ng。然后，开发并优化了物种特异性的多重PCR方法，以同时检测绵羊/山羊（237 bp），牛（223 bp），一次反应中，鸡（192 bp），鸭（168 bp）和猪（154 bp）。通过开发的多重PCR方法检测了包含一种或多种动物来源成分的各种加工肉制品，其结果与其标记的肉类一致。我们的研究为调节肉制品中动物源成分的标签提供了一种快速简便的检测方法。