可视化-跨越式滚环等温扩增技术检测鼠肉掺假

Determination of rat meat adulteration by visualization-saltatory rolling circle isothermal amplification technology

中文关键词:  跨越式滚环等温扩增技术  鼠肉  可视化  掺假

英文关键词:saltatory rolling circle isothermal amplification technology  rat meat  visualization  adulteration

中文摘要:

      目的 建立可视化-跨越式滚环等温扩增技术检测肉制品中鼠肉掺假的分析方法。方法 选取鼠的线粒体cyt b基因为靶基因, 设计并筛选出一对引物, 选择9个不同物种的新鲜肌肉组织样本为研究对象, 验证跨越式滚环等温扩增技术(saltatory rolling circle isothermal amplification, SRCA)方法的特异性; 测定该技术的灵敏度以及人工污染样品中鼠肉成分的检出限, 验证SRCA方法的准确性。结果 SRCA荧光可视化法检测鼠肉DNA的灵敏度为7.3×100 fg/μL, 与传统PCR方法相比, SRCA荧光可视化法的灵敏度提高了1000倍。在人工添加鼠肉的模拟掺假样品中, SRCA方法可检测到含量为0.01%的鼠肉。结论 SRCA技术可以灵敏、快速、准确地检测出肉制品中的鼠肉掺假成分, 适用于基层单位对肉制品掺假的快速检测。

英文摘要:

      Objective To establish a method for detection the rat meat adulteration in meat products by visualized-saltatory rolling ring isothermal amplification technique. Methods The mouse mitochondrial gene cyt b was selected as the target gene, and a pair of primers were designed and screened. Nine different species fresh muscle tissue samples were selected as experimental objects to verify the replacement of the saltatory rolling circle isothermal amplification (SRCA) method. The sensitivity of the technique and the limit of detection of rat meat components in contaminated samples were measured to verify the accuracy of the SRCA method. Results The sensitivity of SRCA fluorescence visual method to detect rat meat DNA was 7.3×100 fg/μL, and compared with the traditional PCR method, the sensitivity of the SRCA fluorescence visualization method was increased by 1000 times. In the simulated repeated fake samples of artificially added rat meat, the content of rat meat detected by SRCA method was 0.01%. Conclusion The SRCA technology established in this study can sensitively, quickly and accurately detect the composition of rat meat penetrating fake meat products, and it is suitable for the rapid detection of adulteration in meat products by grassroots units.