摘要
基于磁分离富集核酸适配体技术,构建了一种新型比色型适配体传感器,用于快速检测大田软海绵酸。巯基修饰的固定链自组装到磁珠上,与适配体链杂交反应后,再加入带有HRP-纳米金标记的探针链与适配体链的3端结合。从而形成一个带有HRP-纳米金标记的适配体传感器,催化TMB产生变色反应,测定在A610/A520的值。在最优条件下,传感器具有良好的线性检测范围(0.2~2.0 ng·mL^(-1))和较高的灵敏度,R^(2)=0.9989,检测限(LOD)0.044 ng·mL^(-1),在加标回收实验中时,平均回收率为82.23%~98.73%,相对标准偏差RSD均小于11%,本研究成功构建了一种用于大田软海绵酸快速、高灵敏度的显色型适配体生物传感器,为大田软海绵酸的快速检测提供了新的思路和研究基础。
In this study,based on the enrichment of nucleic acid aptamers by magnetic separation,a novel colorimetric aptamer sensor was constructed for the rapid detection of okadaic acid.The sulfhydryl-modified fixed chain is self-assembled on the magnetic beads,and after hybridization reaction with the aptamer chain,the probe chain labeled with HRP-nano-gold is added to bind to the 3 ends of the aptamer chain.Thus,an aptamer sensor with HRP-nano-gold labeling was formed,which catalyzed TMB to produce a color-changing reaction,and the value at A610/A520 was determined.Under optimal conditions,the sensor has a good linear detection range(0.2-2.0 ng·mL^(-1))and high sensitivity,R^(2)=0.9989,and the limit of detection(LOD)is 0.044 ng·mL^(-1).In spike recovery experiments,the average recovery rate was 82.23%-98.73%,and therelative standard deviation(RSD)was less than 11%.This experiment successfully constructed a chromogenic aptamer sensor for the efficient and rapid detection of okadaic acid,and provided a new idea and research basis for the rapid detection of okadaic acid.
作者
王晓煜
劳敏军
胡慧利
周勇
吴益春
邓尚贵
胡艺
于瑾
WANG Xiao-yu;LAO Min-Jun;HU Hui-li(School of Food and Pharmacy of Zhejiang Ocean University,Zhoushan 316022;Zhejiang Xingye Group Co Ltd,Zhoushan 316000;Hangzhou Quality and Technology Association,Hangzhou 310019,China)
出处
《浙江海洋大学学报（自然科学版）》
CAS
2022年第2期136-142,共7页
Journal of Zhejiang Ocean University：Natural Science
基金
国家重点研发计划“蓝色粮仓科技创新”重点专项(2020YFD0900900)
浙江省基础公益研究计划项目(LGN18C200023)。
