基于线粒体DNACOⅠ基因鉴别畜禽肉中鸡源性成分.PDF

44 2017, Vol. 31, No. 08 肉类研究 中国肉类食品综合研究中心 分析检测 MEAT RESEARCH CHINA MEAT RESEARCH CENTER 基于线粒体DNA CO Ⅰ基因鉴别畜禽肉中 鸡源性成分 唐修君，樊艳凤，葛庆联，贾晓旭，张 静，刘茵茵，张小燕，王 珏，高玉时* （中国农业科学院家禽研究所，江苏扬州 225125 ） 摘  要：为建立畜禽肉中鸡源性成分的快速检测方法，以线粒体DNA细胞色素C氧化酶亚基Ⅰ（CO Ⅰ）基因序列 为靶点，比较羊、牛、猪、兔、鸽、鹌鹑、鸡、鸭、鹅9 种动物CO Ⅰ基因的差异位点，设计筛选鸡特异性引物， 进行常规聚合酶链式反应（polymerase chain reaction ，PCR ）和荧光定量PCR检测。结果表明：所设计的引物仅对 鸡肉DNA模板有扩增条带和典型扩增曲线，且循环阈（cycle threshold ，Ct ）值为21.78 ，对其他动物DNA模板无扩 增。方法特异性较强，灵敏度较高，达pg级，可以用于畜禽肉与肉制品中鸡源性成分的快速、有效、准确检测。 关键词：鸡源性成分；线粒体DNA ；CO Ⅰ基因；荧光定量PCR Identification of Chicken Origin Ingredients in Livestock and Poultry Meat Based on Mitochondrial DNA COI Gene * TANG Xiujun, FAN Yanfeng, GE Qinglian, JIA Xiaoxu, ZHANF Jing, LIU Yinyin, ZHANG Xiaoyan, WANG Jue, GAO Yushi (Institute of Poultry, Chinese Academy of Agricultural Sciences, Yangzhou 225125, China) Abstract: In this study, a quick method to identify ingredients of chicken origin in livestock and poultry meat was developed by polymerase chain reaction (PCR) and fluorescence quantitative PCR using mitochondrial DNA cytochrome C coxidase subunit Ⅰ (CO Ⅰ) gene as target gene. The mitochondrial DNA CO Ⅰ genes from chicken, duck, goose, pig, cattle, sheep, rabbit, pigeon and quail revealed distinct loci. Based on this, a set of chicken-specific primers was designed. The results showed that the designed primers could only amplify template DNA from chicken with a cycle threshold (Ct) value of 21.78. The PCR assay had a high specificity and sensitivity (at pg level) and it could be used to detect ingredients of