羊源性基因组DNA标准物质研制  

Development of Sheep-derived Genomic DNA Reference Material

作　　者：纪艺 王凯莉 余卉茹 赵新[4] 丁霖 彭城 徐俊锋[1] 陈笑芸[1] JI Yi;WANG Kaili;YU Huiru;ZHAO Xin;DING Lin;PENG Cheng;XU Junfeng;CHEN Xiaoyun(State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products,Key Laboratory of Traceability for Agricultural Genetically Modified Organisms,Ministry of Agriculture and Rural Affairs,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China;School of Food and Pharmacy,Ningbo University,Zhejiang Ningbo 215211,China;Xianghu Laboratory,Hangzhou 311231,China;Institute of Germplasm Resources and Biotechnology,Tianjin Academy of Agricultural Sciences,Tianjin 300384,China)

机构地区：[1]浙江省农业科学院,农产品质量安全危害因子与风险防控国家重点实验室,农业农村部农业转基因生物溯源重点实验室,杭州310021 [2]宁波大学食品与药学学院,浙江宁波215211 [3]湘湖实验室,杭州311231 [4]天津市农业科学院,种质资源与生物技术研究所,天津300384

出　　处：《生物技术进展》2024年第1期125-132,共8页Current Biotechnology

基　　金：科技创新专项2030重大项目农业生物育种重大专项(2022ZD0402012);浙江省重点研发项目(2021C02059)。

摘　　要：近年来,羊肉及其肉制品的掺假等食品安全事件层出不穷,为了完善市场执法检查法律依据,利用数字PCR定值羊HELZ基因,研制了羊基因组DNA标准物质。由于标准物质可以用于衡量检测方法的准确性,因此可以判定食品及相关制品中羊肉的掺假情况。利用数字PCR对研制的标准物质进行均匀性和稳定性评估,结果表明该批标准物质均匀性良好,在4℃、25℃可以稳定保存14 d,在-20℃可以稳定保存6个月。来自全国9个不同实验室羊源性基因组DNA标准物质(高浓度)和羊源性基因组DNA标准物质(低浓度)的联合定值结果显示,标准值及其扩展不确定度分别为(5.44±0.45)×10^(3)copies·μL^(-1)和(5.68±0.54)×10^(2)copies·μL^(-1)。该羊源性基因组DNA标准物质为动物源性标准物质的市场应用提供了技术基础,完善了羊源性基因组DNA标准物质的制备、定量检测、质量控制和量值溯源技术平台。In recent years,food safety incidents such as adulteration of mutton and its meat products have been occurring,and in order to improve the legal basis for market enforcement and inspection,a standard substance for sheep genomic DNA has been developed by using digital PCR to quantify the sheep HELZ gene.Since the standard substance can be used to measure the accu⁃racy of detection methods,it can determine the adulteration of mutton in food and related products.In this research,the standard material of sheep genome DNA was developed as the positive control for the detection of mutton in food and related products.The uniformity and stability of the reference materials were evaluated by digital PCR.The results showed that the reference materials had good uniformity and could be stably stored at 4℃and 25℃for 14 days,and at-20℃for 6 months.The standard values of sheep-derived genomic DNA reference material(high concentration)and sheep-derived DNA reference material(low concentra⁃tion)and their extended uncertainties were(5.44±0.45)×10^(3) copies·μL^(-1) and(5.68±0.54)×10^(2) copies·μL^(-1),respectively, which were jointly determined by nine different laboratories. The development of sheep-derived genomic DNA reference materials pro⁃ vides a technical basis for the development of animal-derived reference materials, and will establish a technical platform for the prepa⁃ ration, quantitative detection, quality control and quantitative value traceability of sheep-derived genomic DNA reference materials.

关 键 词：标准物质 羊源性HELZ基因 数字PCR 

分 类 号：Q789[生物学—分子生物学] TS201.6[轻工技术与工程—食品科学]