Enzymatically amplified surface plasmon resonance imaging detection of DNA by exonuclease III digestion of DNA microarrays
Enzymatically amplified surface plasmon resonance imaging detection of DNA by exonuclease III digestion of DNA microarrays
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DOI：
10.1021/ac050815w
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发表时间：
2005-08-15
影响因子：
7.4
通讯作者：
Corn, RM
中科院分区：
文献类型：
--
作者：
Lee, HJ;Li, Y;Corn, RM
本文介绍了一种新的方法，利用酶核酸外切酶III结合T-终止的DNA微阵列的扩增检测单链DNA（ssDNA）的表面等离子体共振（SPR）成像。当ExoIII和靶DNA同时引入T-终止的ssDNA微阵列时，靶ssDNA的杂交吸附导致探针ssDNA链的方向依赖性ExoIII水解和完整靶ssDNA释放回溶液中。靶ssDNA与另一探针的再吸附产生重复的水解过程，其随时间推移导致SPR成像信号的显著负变化。实验证明ExoIII与双链DNA的方向依赖性表面酶反应，以及这种新的酶促扩增SPR成像过程与16-mer靶ssDNA检测限为10-100 pM。这是对先前报道的仅基于杂交吸附而没有酶促扩增的ssDNA的SPR成像检测的测量的10（2）-10（3）改进。
This paper describes a novel approach utilizing the enzyme exonuclease III in conjunction with T-terminated DNA microarrays for the amplified detection of single-stranded DNA (ssDNA) with surface plasmon resonance (SPR) imaging. When ExoIII and target DNA are simultaneously introduced to a T-terminated ssDNA microarray, hybridization adsorption of the target ssDNA leads to the direction-dependent ExoIII hydrolysis of probe ssDNA strands and the release of the intact target ssDNA back into the solution. Readsorption of the target ssDNA to another probe creates a repeated hydrolysis process that results over time in a significant negative change in SPR imaging signal. Experiments are presented that demonstrate the direction-dependent surface enzyme reaction of ExoIII with double-stranded DNA as well as this new enzymatically amplified SPR imaging process with a 16-mer target ssDNA detection limit of 10-100 pM. This is a 10(2)-10(3) improvement on previously reported measurements of SPR imaging detection of ssDNA based solely on hybridization adsorption without enzymatic amplification.