常见肉类中鸭源性成分荧光PCR检测方法的建立.pdfVIP

第 7 卷第 2 期 食品安全质量检测学报 Vol. 7 NO.2 2016 年 2 月 Journal ofFood Safety and Quality Feb. , 2016 常见肉类中鸭源性成分荧光 PCR 检测方法的建立 王佳慧，吴爽，李楠，宋忠佳，王晓菲，李风琴，江涛* (国家食品安全风险评估中心，北京 100021) 摘要:目的通过筛选引物、探针体系优化 PCR 反应条件建立常见肉品中鸭源性 DNA 的荧光 PCR 检测 方法。方法针对线粒体基因设计划对引物探针以鸭 DNA 为模板利用 PCR 和荧光 PCR 方法筛选引物探 针并确定反应条件，进行灵敏度、特异性和稳定性试验，最终确定荧光定量 PCR 方法的引物、探针及反应条件。 结果 当样本含量为 0.1%时，本方法仍可特异、灵敏地检出鸭肉成分，检测 Ct 值可稳定在 20 以下。结论建 立了常见肉品中鸭源性 DNA 的荧光 PCR 检测方法可用于肉类制品中鸭源性成分的鉴定。 关键词:实时荧光定量 PCR; 鸭肉;线粒体 DNA Establishment of real-time fluorescence quantitative PCR method for identifying duck ingredients from edible meat WANG Jia-Hui, WU Shuang, LI Nan, SONG Si-Jia, WANG Xiao-Fei, LI Feng-Qin, JIANG Tao拿 (China National Center for Food Safety RiskAssessment, Beijing 100021, China) ABSTRACT: Objective To establish a real-time fluorescence quantitative PCR method for identifying the hidden duck ingredients in other meats through screening of primers and probe system and optimizing of PCR reaction conditions. Methods Totally fi 企y primer pairs and probes were designed according to sequence of the cytochrome b gene 仕om the duck. Primers and probe were selected and the conditions of the PCR were confirmed. Specificity and sensitivity of the method were evaluated. Results Duck ingredients could be identified specifically and sensitively, even the content percentage was 0.1 %. The Ct value was below 20. Conclusion The real-time fluorescence quantitative PCR method for identification of duck has h