基于RPA技术对转CP4-EPSPS基因产品的快速检测  

Rapid Detection of CP4-EPSPS Transgenic Products Based on RPA Technology

作　　者：胡秀文 邓波 王金斌[3] 刘华[3] 唐雪明 王宇 曾海娟 蒋玮[3] 李红[6] HU Xiuwen;DENG Bo;WANG Jinbin;LIU Hua;TANG Xueming;WANG Yu;ZENG Haijuan;JIANG Wei;LI Hong(College of Food Science and Technology,Shanghai Ocean University,Shanghai 201306,China;Shanghai Center of Agriproducts Quality and Safety,Shanghai 201708,China;Biotech Research Institute,Shanghai Academy of Agricultural Sciences,Shanghai 201106,China;Shanghai Jiao Tong University School of Agriculture and Biology,Shanghai 200240,China;School of Life Science and Engineering,Lanzhou University of Technology,Lanzhou 730050,China;Institute of Animal Husbandry&Veterinary Science,Shanghai Academy of Agricultural Sciences,Shanghai 201106,China)

机构地区：[1]上海海洋大学食品学院,上海201306 [2]上海市农产品质量安全中心,上海201708 [3]上海农业科学院生物技术研究所,上海201106 [4]上海交通大学农业与生物学院,上海200240 [5]兰州理工大学生命科学与工程学院,兰州730050 [6]上海市农业科学院畜牧兽医研究所,上海201106

出　　处：《中国农业科技导报》2023年第2期227-233,共7页Journal of Agricultural Science and Technology

基　　金：上海市科技计划项目(21N31900800);上海市青年科技英才扬帆计划项目(20YF1443000);上海市科委自然科学基金项目(19ZR1436800);上海市农业科学院卓越团队计划项目(农科创2017-B-07)。

摘　　要：核酸检测在农产品安全检测中的应用十分广泛,随着转基因作物的商业化种植,迫切需要一种快速、特异、灵敏的转基因作物检测方法。利用重组酶聚合酶扩增(recombinase polymerase amplification,RPA)技术对含有转基因产品及其制品中CP4-EPSPS基因进行检测,共设计5对引物来筛取最佳引物,并对反应体系和反应温度进行优化。结果表明,该方法采用20μL体系在37℃恒温反应15 min即可对CP4-EPSPS基因进行快速检测。该方法检测转基因的灵敏度阈值为45拷贝,灵敏度高、特异性强,为大规模筛查CP4-EPSPS基因提供了一种新的途径。Nucleic acid detection is widely used in the safety detection of agricultural products.With the commercial cultivation of genetically modified crops,the method with rapid,specific and sensitive is an urgent need for the detection of genetically modified crops.The recombinase polymerase amplification(RPA)technology was used to detect CP4-EPSPS genes in genetically modified products and their products.Five pairs of primers were designed to screen the best pair of primers,and the reaction system and temperature were optimized.The results show that CP4-EPSPS gene could be quickly detected by this method with 20μL reaction system,15 min reacted at 37℃.The sensitivity threshold for detecting transgenic DNA was 45 copies with highly sensitive and specific nucleic,which provided a new way for large-scale screening of CP4-EPSPS gene.

关 键 词：重组聚合酶扩增 CP4-EPSPS 转基因作物 核酸检测 

分 类 号：Q-33[生物学]