基于纳米金标记-适配体识别的伏马菌素B1可视化检测新方法.docVIP

  基于纳米金标记-适配体识别的伏马菌素 B1 可视化检测新方法# 王文凤1，吴世嘉1，谭贵良2，王周平1** 5 10 15 20 25 30 35 40 （1. 江南大学食品学院，江南大学食品科学与技术国家重点实验室，江苏 无锡 214122； 2. 中山市质量计量监督检测所，广东 中山 528403） 摘要：本研究基于核酸适配体识别和纳米金变色效应构建了伏马菌素 B1（FB1）的可视化检 测新方法。实验以纳米金为载体，首先在纳米金表面组装巯基化的适配体互补短链 DNA1/FB1-适配体复合物；当加入目标物时，适配体链与目标物结合，与互补短链 DNA1 发生 解离；此时再加入纳米金标记的与适配体互补短链 1 互补的短链 DNA 2，二者杂交可导致纳 米金粒子的聚集而使溶液颜色发生变化，进而实现目标物的可视化检测。通过条件优化，有 效避免了由于盐浓度过大使纳米金发生聚集所产生的误差。同时在纳米金与短链 DNA 孵化时 加入表面活性剂十二烷基硫酸钠（SDS），使 Nacl 浓度达到了 500 mmol/L 而纳米金颜色仍 不发生改变，打破了以往熟化 Nacl 浓度 100 mmol/L 就使纳米金颜色发生变化的界限，使附 着在纳米金上的 DNA 量扩大了 3 倍。方法检测线性范围为 125-1500 ng/L，检测限为 125 ng/L。 该方法已成功应用于啤酒中 FB1 的检测。 关键词：适配体识别；纳米金；可视化；伏马菌素 B1 Novel naked-eye detection method for Fumonisins B1 detection based on AuNPs color changes and aptamer recognition Wang Wenfeng1, Wu Shijia1, Tan Guiliang2, Wang Zhouping1 (1. State Key Laboratory of Food Science and Technology,School of Food Science and Technology,Jiangnan University, JiangSu WuXi 214122; 2. Zhongshan Supervision Testing Institute of Quanlity and Metrology, GuangDong ZhongShan 528403) Abstract: This work developed a new naked eye detection method for FB1 abased on the high identification function of DNA aptamer and the color changing effect of AuNPs. This method taked AuNPs as carrier and linked DNA1 to it firstly and then assembled FB1-aptamer -DNA1-AuNPs complex throuth the hybridzation process between FB1-aptamer and DNA1-AuNPs. When added different concentrations of FB1, FB1-aptamer prefered to connect to FB1 and then subsequently additional DNA2 chain that matched with DNA 1chain can hybrid with exposed DNA1 resulting in AuNPs aggregation.Thus the color changes from red to purple and blue could be observed by the naked eye. Optimal conditions avoided to much salt induced AuNPs aggregation caused error effectively. Furthermore, we added SDS in DNA1 or DNA2 chain connected to AuNPs to maintain the color of AuNPs and improved the concentration of Nacl to 500 mmol/L and DNA loading enlarge 3 times. This breaks the traditional bo