基于环介导等温扩增快速检测转基因大豆SHZD32-1成分  

Rapid Detection of Transgenic Soybean SHZD32-1 by Loop-mediated Isothermal Amplification

作　　者：邢宇俊[1] 梁杰 王亚林 吴季荣[1] 董飞[1] 史建荣[1] 徐剑宏[1] 梁晋刚 XING Yu-jun;LIANG Jie;WANG Ya-lin;WU Ji-rong;DONG Fei;SHI Jian-rong;XU Jian-hong;LIANG Jin-gang(Institute of Food Safety and Nutrition,Jiangsu Academy of Agricultural Sciences/Jiangsu Provincial Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base Built by Province and Ministry,Ministry of Science and Technology/Jiangsu Center for GMO Evaluation and Detection,Nanjing 210014,China;School of Life Science and Technology,Sanquan College of Xinxiang Medical University,Xinxiang 453002,China;Development Center of Science and Technology,Ministry of Agriculture and Rural Affairs,Beijing 100176,China)

机构地区：[1]江苏省农业科学院农产品质量安全与营养研究所/江苏省食品质量安全重点实验室-省部共建国家重点实验室培育基地/江苏省转基因安全评价公共服务中心,江苏南京210014 [2]新乡医学院三全学院,生命科学技术学院,河南新乡4530022 [3]农业农村部科技发展中心,北京100176

出　　处：《大豆科学》2023年第1期41-47,共7页Soybean Science

基　　金：国家自然科学基金(31301488);江苏省农业科技自主创新资金[CX(22)3072]。

摘　　要：转基因大豆SHZD32-1是由我国自主研发的耐除草剂抗性品系。为给该品系转基因生物安全监管提供技术支撑,本研究开发其转基因成分的快速便捷式检测方法,主要基于环介导等温扩增技术,设计并筛选该转化体最佳特异性LAMP引物组,优化扩增温度和时间,并分析灵敏度、特异性及稳定性,建立SHZD32-1的LAMP检测体系。结果显示:在62~64℃条件下反应40~60 min扩增效果最佳;灵敏度标准达到质量分数0.05%,高于普通PCR(0.1%);特异性和稳定性较高,检测结果同普通PCR扩增一致。转基因大豆SHZD32-1成分LAMP技术不需要特殊仪器,检测时间短,结果可视化,灵敏度、特异性和稳定性高,有望用于转基因耐草甘膦大豆SHZD32-1的现场快速检测。Transgenic soybean SHZD32-1 is a transgenic material developed by Chinese scientists. In order to provide technical support for the safety supervision of the material, this study developed a rapid and convenient method for detecting the transgenic soybean SHZD32-1, which was based on loop-mediated isothermal amplification. We designed and screened the optimal specific LAMP primer, optimized the amplification temperature and time, and analyzed the sensitivity, specificity and stability. The results showed that the optimal conditions were 62-64 ℃ for 40-60 min. The sensitivity of this LAMP detection system reached 0.05%, more than conventional PCR(0.1%), was higher specificity and stability, and the detection results were consistent with the conventional PCR amplification. The LAMP detection system of transgenic soybean SHZD32-1 doesn’t need special instruments, time-saved, and result visualization with high sensitivity, specificity and stability, and is expected to be used for rapid detection of transgenic glyphosate-tolerant soybean SHZD32-1 in the field.

关 键 词：环介导等温扩增(LAMP) 转基因大豆SHZD32-1 基因特异性 快捷检测 

分 类 号：S565.1[农业科学—作物学]