GNM C7-8实时荧光PCR同时快速检测8种动物源性成分

目的基于八模块设计的GNMC7-8实时荧光PCR系统实现对8种动物源性成分同时快速检测。方法基于猪朊蛋白基因、牛生长激素基因、羊生长激素基因、驴线粒体ATPase 6基因、马线粒体ATPase 6基因、鸭生长激素基因、水貂线粒体细胞色素b基因、狐狸线粒体细胞色素氧化酶Ⅰ亚基基因引物设计了8种动物源性成分实时荧光PCR检测方法。应用GNMC7-8实时荧光PCR系统,对8种动物源性成分启动同时检测和每隔5min顺序启动不同时检测,并将检测结果和ABI7500荧光PCR分别检测结果进行比较。结果 GNM C7-8实时荧光PCR系统同时检测猪、牛、羊、驴、马、鸭、水貂、狐狸8种动物源性成分,起峰周期数分别为30、27、21、21、24、17、15、17,与其不同时检测结果一样,与ABI7500荧光PCR系统检测结果相比,猪、羊、马、鸭、水貂5种成分的起峰快1～2个周期数,其余3种成分检测结果一致,检测完成时间少16～20 min。结论 GNM C7-8实时荧光PCR系统8个模块独立运行,互不干扰,无交叉污染,能够同时快速检测8种动物源性成分,为快速筛查动物源性成分提供强有力的设备技术支持。

Simultaneous rapid detection of 8 kinds of animal-derived ingredients by GNM C7-8 real-time PCR method

Objective To establish a method for simultaneous rapid detection of 8 kinds of animal-derived ingredients by GNM C7-8 real-time PCR system with 8 modules. Methods Based on porcine prion gene primers, bovine growth hormone gene, ovine growth hormone gene, donkey mitochondrial ATPase 6 gene, horse mitochondrial ATPase 6 gene, duck growth hormone gene, mink mitochondrial cytochrome b gene, fox mitochondrial cytochrome oxidase I subunit gene, 8 kinds of real-time PCR detection methods of animal derived components were designed. GNM C7-8 real-time fluorescence PCR system was used to initiate simultaneous detection of 8 animal-derived components and sequential initiation of different time detection at 5 min intervals, and the results were compared with those of ABI 7500 fluorescent PCR. Results GNM C7-8 real-time fluorescence PCR system was used for the simultaneously detection of 8 animal origin components of pig, cow, sheep, donkey, horse, duck, mink and fox, and the number of peak cycles was 30, 27, 21, 21, 24, 17, 15, 17, respectively, which were the same as the detection results at different times. Compared with the detection results of ABI 7500 fluorescence PCR system, the peaking of the 5 components of pig, sheep, horse, duck and mink was 1~2 cycles faster, the detection results of the other 3 components were consistent, and the detection completion time was 16~20 min less. Conclusions The GNM C7-8 real-time fluorescent PCR system operates independently of each other, without interference, without cross-contamination, and can quickly detect 8 animal-derived components at the same time, which provides powerful equipment technical support for rapid screening of animal-derived components.