基于DNAzyme和杂交链式反应用于检测铅离子的荧光生物传感器研究  

Study on a fluorescent biosensor based on DNAzyme and hybrid chain reaction for detection of lead ions

作　　者：汪小坤 易怀超 林峰仪 顾荣梦 聂泓宇 李志慧 戴建远[2] 袁红雁[1] WANG Xiao-kun;YI Huai-chao;LIN Feng-yi;GU Rong-meng;NIE Hong-yu;LI Zhi-hui;DAI Jian-yuan;YUAN Hong-yan(College of Chemistry Engineering,Sichuan University,Chengdu,610065,China;College of Chemical,Sichuan University,Chengdu,610064,China)

机构地区：[1]四川大学化学工程学院,四川成都610065 [2]四川大学化学学院,四川成都610064

出　　处：《化学研究与应用》2022年第12期2943-2949,共7页Chemical Research and Application

基　　金：四川省农业科学院分析测试中心合作项目(20H1090、0020303410001)资助。

摘　　要：通过将GR-5 DNAzyme的序列设计进发生杂交链式反应的发夹DNA中,在引发DNA的触发下,发夹DNA H1与H2反应生成具有重复双螺旋结构的纳米线T-H1-H2-H1-H2…。在H1与H2结合部分存在碱基不配对区域,此处将形成GR-5 DNAzyme结构,在铅离子的存在下,该部分GR-5 DNAzyme的底物链被裂解,HCR产物即被裂解成许多DNA双链小分子从而实现信号扩增,以此构建了简便灵敏的荧光生物传感器。该生物传感器可在30min完成对Pb的检测,线性范围为0.1μM~7μM,检测限为23.2nM。对实际水样进行检测,加标回收率在96.7%~106.6%之间,相对标准偏差均小于2%。A simple and sensitive biosensor was constructed by incorporating GR-5 DNAzyme sequence into hairpin DNA sequence for hybridization chain reaction.Under the trigger of trigger DNA,hairpin DNA H1 reacts with H2 to generate nanowires T-H1-H2-H1-H2...with repetitive double helix structure.There is a base unpaired region in H1 and H2 binding part,where the GR-5 DNAzyme structure is formed.In the presence of lead ions,the substrate chain of this part of GR-5 DNAzyme is cleaved,so that the HCR product are cleaved into many DNA double-stranded small molecules which caused signal amplification,and thus constructed a simple and sensitive fluorescent biosensor.The biosensor could detect Pbwithin 30 min with a linear range of 0.1μM to 7μM and a detection limit of 23.2 nM.This biosensor was used for Pbdetection in actual water samples and the recoveries ranged from 96.7%to 106.6%with relative standard deviations less than 2%.

关 键 词：GR-5 DNAzyme 杂交链式反应 荧光生物传感器 铅离子检测 

分 类 号：O657.3[理学—分析化学]