基于改进G-四链体DNA酶的电化学适配体传感器构建及卡那霉素高灵敏检测  

Construction of electrochemical aptasensor for highly sensitive detection of kanamycin based on the improved G-quadruplex DNAzyme

作　　者：伍永梅 朱肖倩 方娇 白艳红 WU Yongmei;ZHU Xiaoqian;FANG Jiao;BAI Yanhong(College of Food and Bioengineering,Zhengzhou University of Light Industry,Zhengzhou 450001,China;Henan Key Laboratory of Cold Chain Food Quality and Safety Control,Zhengzhou 450001,China;Collaborative Innovation Center of Food Production and Safety of Henan Province,Zhengzhou 450001,China)

机构地区：[1]郑州轻工业大学食品与生物工程学院,河南郑州450001 [2]河南省冷链食品质量安全控制重点实验室,河南郑州450001 [3]食品生产与安全河南省协同创新中心,河南郑州450001

出　　处：《轻工学报》2023年第6期62-69,共8页Journal of Light Industry

基　　金：河南省重点研发与推广专项项目(222102310140);河南省高等学校重点科研项目(21A550015);郑州轻工业大学博士启动科研基金项目(0123-13501050070)。

摘　　要：将阳离子多肽(peptide)与G-四链体DNA酶(G4 DNAzyme)共价组装得到高活性的G4 DNAzyme-peptide复合物,进一步构建新型电化学适配体传感器,研究该复合物对电化学适配体传感器响应性能的影响,并将该传感器用于牛奶中卡那霉素(KANA)的检测分析。结果表明:G4 DNAzyme-peptide可以显著放大电化学信号,明显提高传感器检测灵敏度;在捕获探针序列最优浓度(2.0μmol/L)下,电流信号强度变化与目标物浓度(0.06 pmol/L~20 nmol/L)对数值呈良好的线性关系,检测限为0.02 pmol/L,优于其他KANA检测方法;该传感器对KANA具有良好的选择性,在实际牛奶样品检测中,KANA的加标回收率为97.1%~105.5%,相对标准差为3.60%~5.74%,且检测结果与ELISA法一致,说明该传感器能够实现牛奶中KANA的高灵敏检测。A highly active G4 DNAzyme-peptide complex obtained by covalent assembly of cationic peptide on G4 DNAzyme was used to construct a novel electrochemical aptasensor.The effect of complex on the performance of electrochemical aptasensor was investigated and the proposed aptasensor was used to detect kanamycin(KANA)in milk.The experimental results indicated that G4 DNAzyme-peptide could significantly amplify the electrochemical signal and greatly improve the detection sensitivity of aptasensor.Under the optimal concentration of capture probe(CP,2.0μmol/L),the change of current intensity showed a good linear relationship with the logarithm of the target concentration in the range of 0.06 pmol/L~20 nmol/L,and the detection limit was 0.02 pmol/L,which was superior to other KANA detection methods.The aptasensor showed better selectivity and lower detection limit and the recoveries obtained in milk sample were from 97.1% to 105.5%and the relative standard deviation were from 3.60% to 5.74%.The experimental results were consistent with ELISA method,indicating the proposed aptasensor could achieve high sensitivity for the detection of KANA in milk.

关 键 词：G-四链体DNA酶 电化学适配体传感器 卡那霉素 高灵敏度 

分 类 号：TS207.3[轻工技术与工程—食品科学]