实时重组酶辅助扩增技术快速检测狗源性成分  

Rapid Detection of Dog-Derived Ingredients by Real-Time Recombinase-Aided Amplification

作　　者：刘明明 颜海燕 周朝旭 叶语桦 李志刚 肖昭竞 LIU Mingming;YAN Haiyan;ZHOU Zhaoxu;YE Yuhua;LI Zhigang;XIAO Zhaojing(Chongqing Academy of Metrology and Quality Inspection(National Center of Quality Supervision&Inspection of Agricultural Processed Products and Condiments),Chongqing 401123,China)

机构地区：[1]重庆市计量质量检测研究院(国家农副加工产品及调味品质量检验检测中心),重庆401123

出　　处：《肉类研究》2025年第4期24-29,共6页Meat Research

基　　金：重庆市市场监督管理局科研计划项目(CQSJKJDW2023006)。

摘　　要：为快速检测狗源性成分,针对cytb基因保守区域设计3对重组酶辅助扩增(recombinaseaided amplification,RAA)引物和1条核酸外切酶(exonuclease,exo)探针,通过引物和探针浓度的筛选,构建一种实时RAA(real-time RAA,Rt-RAA)方法。利用交叉反应和重复性实验对该检测方法的特异性和稳定性进行评估,并将Rt-RAA方法灵敏度以及对混合模拟样品和市售样品的检测结果与相应的国标检测方法进行对比。结果表明,Rt-RAA检测方法特异性强、稳定性好,在30 min内可检测到目的序列,对狗源性成分的检出限为0.2%,明显优于实时荧光聚合酶链式反应检测方法,应用Rt-RAA方法与国标方法对模拟样品和市售实际样品检测结果完全一致。因此,本研究建立的Rt-RAA检测方法适用于狗源性成分的检测,为肉源性成分鉴定提供了另一种快速、准确的新方法。In order to establish a real-time recombinase aided amplification(Rt-RAA)method to rapidly detect dog-derived ingredients,three pairs of primers and one exonuclease(exo)probe were designed targeting the conserved region of the cytochrome B(cytb)gene.The designed primer combinations and probe concentration were optimized.The specificity and stability of the method were evaluated by cross-reactivity and repeatability experiments.The sensitivity of the Rt-RAA method and its results for simulated mixed samples and commercial samples were compared with those of the national standard method.The results showed that the Rt-RAA method had strong specificity and good stability and could detect the target sequence in less than 30 min.The detection limit for dog-derived ingredients was 0.2%,which was significantly better than that of real-time fluorescence polymerase chain reaction(PCR).The results of the Rt-RAA method for simulated mixed samples and commercial samples were completely consistent with those of the national standard method.Therefore,Rt-RAA is suitable for the detection of dog-derived ingredients,providing a fast and accurate new method for the identification of meat-derived ingredients.

关 键 词：实时荧光重组酶辅助扩增技术 狗源性成分 肉制品掺假 

分 类 号：TS251.7[轻工技术与工程—农产品加工及贮藏工程]