基于磁性辅助的杂交链反应放大检测三磷酸腺苷  

Amplification Detection of Adenosine-Triphosphate Based on Hybrid Chain Reaction and Auxiliary Magnetic Sensing Strategy

作　　者：吕菊波[1] 张亚会[1] 刘刚[1] 徐慧[1] 

机构地区：[1]鲁东大学化学与材料科学学院,烟台264025

出　　处：《化学通报》2018年第1期59-64,76,共7页Chemistry

基　　金：山东省自然科学基金项目(ZR2016BM27);山东省重点研发计划项目(2016GNC111016);烟台市重点研发计划项目(2016ZH059)资助

摘　　要：本文提出了一种基于磁性辅助的杂交链反应放大检测三磷酸腺苷(ATP)的传感策略。磁性纳米粒子表面易于修饰,而且操作方便,具有很好的分离效果,能够提高生物传感的选择性。首先,利用生物素与链霉亲和素之间的亲和力作用,将生物素标记的ATP核酸适配体连接到链霉亲和素修饰的磁性纳米粒子表面,加入与ATP核酸适配体互补的一段DNA进行杂交,通过磁性分离除去未杂交上的DNA,加入靶向ATP,ATP与其适配体特异性结合将适配体的互补链通过磁性分离出来,磁性分离出的信号DNA继续用于下一步的杂交链反应,将信号放大,最后利用氧化石墨烯(GO)对荧光的猝灭效应降低背景荧光,达到高灵敏度、高选择性检测靶向ATP。其中,ATP的最低检测浓度为0.1nmol/L。We put forward a hybrid chain reaction amplification detection of adenosine triphosphate （ATP） basdd on auxiliary magnetic sensing strategy. The surface of magnetic nanoparticles is easy to be modified, and they have the advantages of the convenient operation, the good separation effect and the higher selectivity for biological sensing. First of all, the biotin labeled ATP aptamer will connect to the surface of the magnetic nanoparticles which is modified with streptavidin based on the affinity between biotin and streptavidin. Then a single DNA which is complementary to the ATP aptamer is added to hybrid with ATP aptamer, and the unnecessary DNA will be removed by magnetic separation. When the targeted ATP is added to the solution, the ATP will bind with its aptamer specially by releasing the complementary single-stranded DNA, and the complementary single-stranded DNA will continue to be used for the next step of hybridization chain reaction. The signal would be stronger after magnetic separation. The background will be reduced by the fluorescence quenching effect of the graphene oxide, and the method has high sensitivity and high selectivity for targeting ATP. The lowest detection concentration of ATP is 0. 1 nmol/L.

关 键 词：氧化石墨烯 杂交链反应 磁性辅助 荧光信号放大 

分 类 号：O657.3[理学—分析化学] Q503[理学—化学]