Single-base mismatch detection based on charge transduction through DNA
Single-base mismatch detection based on charge transduction through DNA
复制标题
DOI：
10.1093/nar/27.24.4830
复制
发表时间：
1999-12-15
影响因子：
14.9
通讯作者：
Hill, MG
中科院分区：
文献类型：
--
作者：
Kelley, SO;Boon, EM;Hill, MG
对于基因突变和疾病的常规筛查，需要能够检测单碱基错配的高通量DNA传感器。基于DNA膜中的电荷传输，发展了一种检测DNA中单碱基错配的新方法。在金表面制备了双螺旋DNA膜，并用于电化学检测DNA错配。从结合在DNA修饰的金表面上的氧化还原活性嵌入物获得的信号对固定的双链中存在碱基错配具有显着的敏感性。差分错配检测是在不考虑DNA序列组成和错配同一性的情况下完成的。还可以准确地检测在电极表面杂交的序列中的单碱基变化。将插层物种的氧化还原反应与溶液中的电催化过程相结合，大大提高了该分析的灵敏度。与单链寡核苷酸的杂交能量学不同，基于电荷传输的电化学传感器在报告DNA的电子结构方面可能在范围和灵敏度方面具有根本优势。
High-throughput DNA sensors capable of detecting single-base mismatches are required for the routine screening of genetic mutations and disease. A new strategy for the electrochemical detection of single-base mismatches in DNA has been developed based upon charge transport through DNA films. Double-helical DNA films on gold surfaces have been prepared and used to detect DNA mismatches electrochemically. The signals obtained from redox-active intercalators bound to DNA-modified gold surfaces display a marked sensitivity to the presence of base mismatches within the immobilized duplexes. Differential mismatch detection was accomplished irrespective of DNA sequence composition and mismatch identity. Single-base changes in sequences hybridized at the electrode surface are also detected accurately. Coupling the redox reactions of intercalated species to electrocatalytic processes in solution considerably increases the sensitivity of this assay. Reporting on the electronic structure of DNA, as opposed to the hybridization energetics of single-stranded oligonucleotides, electrochemical sensors based on charge transport may offer fundamental advantages in both scope and sensitivity.