肉制品中狗、狐、貂源性成分DNA检测试剂盒的制备

为快速检测肉制品中狗、狐、貂3种常见犬型总科动物源性DNA成分,应用分子生物学技术分析种属基因组序列差异,建立并优化多重聚合酶链式反应检测体系,开发快速检测试剂盒.结果表明:快速检测试剂盒特异性良好,与常见肉类动物(猪、牛、羊、马、驴、鸡、鸭)源性DNA成分皆无交叉反应;灵敏度高,3种靶标样品DNA质量浓度同时降至10...

Development of DNA Detection Kit for Dog,Fox and Mink-Derived Components in Adulterated Meat Products

In order to rapidly detect DNA derived from the common Canidae animals dog, fox and mink in adulterated meat products, molecular biology technology was applied to analyze the differences in the genomic sequence of various species, and a multiplex polymerase chain reaction system was establish and optimized to develop rapid detection kits. The results showed that the developed kit had good specificity without cross-reaction with common food-producing animal-derived DNA components (pigs, cattle, sheep, horses, donkeys, chickens and ducks), as well as high sensitivity. When the DNA concentration of the three target samples decreased to 10-4 ng/μL at the same time, they still could be accurately detected using the kit. The positive bands were excised, cloned and sequenced, showing 100% similarity to the dog (MH105046.1), fox (LT560065.1) and mink (KU145464.1) sequences deposited in the GenBank database. This kit correctly identified 30 simulated mixed meat samples with 100% accuracy. Of 50 commercially available samples, three dog meat samples were found to be adulterated.