基于纳米金/蔗糖酶构建核酸适体传感器检测盐酸多巴胺  

Gold Nanoparticles and Invertase-Based Aptasensor for Detection of Dopamine Hydrochloride

作　　者：姜利英[1,2] 刘帅 任林娇[1] 张培[1] 赵学伟[2,3] 王延峰 JIANG Liying;LIU Shuai;REN Linjiao;ZHANG Pei;ZHAO Xuewei;WANG Yanfeng(College of Electrical and Information Engineering,Zhengzhou University of Light Industry,Zhengzhou 450002,China;Henan Collaborative Innovation Center of Food Production and Safety,Zhengzhou 450002,China;School of Food and Bioengineering,Zhengzhou University of Light Industry,Zhengzhou 450002,China)

机构地区：[1]郑州轻工业学院电气信息工程学院,河南郑州450002 [2]食品生产与安全河南省协同创新中心,河南郑州450002 [3]郑州轻工业学院食品与生物工程学院,河南郑州450002

出　　处：《食品科学》2018年第16期301-305,共5页Food Science

基　　金：国家自然科学基金青年科学基金项目(61002007);河南省科技创新杰出人才项目(184200510015);河南省高等学校重点科研项目(18A510021);河南省高校科技创新团队项目(15IRTSTHN012);郑州轻工业学院博士科研基金项目(2014BSJJ041;2015BSJJ023);郑州轻工业学院研究生科技创新基金项目(2016027)

摘　　要：采用核酸适体做捕获探针,纳米金-蔗糖酶-互补DNA偶合物做检测探针,构建竞争型核酸适体传感器,并结合血糖仪实现对盐酸多巴胺的检测。未加靶目标时,检测探针与核酸适体通过碱基互补配对结合;加入靶目标后,核酸适体与靶目标强亲和力竞争结合,使检测探针脱离捕获探针并催化蔗糖转化为葡萄糖,通过血糖仪检测葡萄糖浓度间接检测盐酸多巴胺浓度。结果表明,在靶目标孵育时间和蔗糖酶催化时间分别为30 min和120 min时,盐酸多巴胺的线性检测范围为0.45~10 mmol/L,相关系数为0.997。对8.0、3.0、0.8 mmol/L盐酸多巴胺标准液进行多次测量,相对标准偏差分别为0.9%、1.4%、4.8%。所制备传感器具有操作简单、稳定性好、选择性高的特点,不仅拓宽了血糖仪对其他非糖标志物的检测,而且在医疗卫生和食品安全检测领域具有应用潜力。By using aptamer as the capture probe and gold nanoparticles-invertase-complementary DNA conjugates as the detection probe,a competitive dopamine hydrochloride(DH)aptasensor was constructed based on a glucose meter.In the absence of DH,the detection probe was captured on a 96-well plate by the aptamer through complementary base pairing.When it was present,DH interacted with its aptamer with a strong affinity forcing the detection probe away from the 96-well plate to catalyze the transformation of sucrose into glucose.Accordingly,indirect detection of dopamine hydrochloride concentration was performed by measuring glucose concentration with a glucose meter.When the incubation time and the enzymatic reaction time were 30 min and 120 min,respectively,the glucose meter signal intensity increase linearly with the increase of DH concentration in the range of 0.45–10 mmol/L with a correlation coefficient of up to 0.997,and the relative standard deviations(RSDs)were 0.9%,1.4%and 4.8%at 8.0,3.0 and 0.8 mmol/L(n=5),respectively.The prepared sensor has the advantages of simple operation,good stability and high selectivity,which not only broadens the application of glucose meter to the detection of other non-glucose,but also has potential application in the fields of health care and food safety detection.

关 键 词：核酸适体 核酸适体传感器 血糖仪 纳米金 盐酸多巴胺 食品安全 

分 类 号：TP212[自动化与计算机技术—检测技术与自动化装置]