Electrochemical DNA biosensor for the detection of short DNA species of Chronic Myelogenous Leukemia by using methylene blue

被引:117
作者
Lin, Xin-Hua
Wu, Ping
Chen, Wei
Zhang, Ya-Feng
Xia, Xing-Hua [1 ]
机构
[1] Nanjing Univ, Key Lab Analyt Chem Life Sci, Sch Chem & Chem Engn, Nanjing 210093, Peoples R China
[2] Fujian Med Univ, Dept Pharmaceut Anal, Fac Pharm, Fuzhou 350004, Peoples R China
基金
中国国家自然科学基金;
关键词
biosensors; DNA; hybridization; CML; methylene blue; sequence-specific electrochemical detection;
D O I
10.1016/j.talanta.2006.11.015
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
A novel assay for the voltammetric detection of 18-bases DNA sequences relating to Chronic Myelogenous Leukemia (CML, Type b3a2) using methylene blue (MB) as the hybridization indicator was reported. DNA was covalently attached onto a glassy carbon electrode (GCE) through amines of the DNA bases using N-hydroxysulfosuccinimide (NHS) and N-(3-dimethylamion)propyl-M-ethyl carbodiimidehydrochloride (EDC). The covalently immobilized single-stranded DNA (ssDNA) could selectively hybridize with its complementary DNA (cDNA) in solution to form double-stranded DNA (dsDNA) on the surface. A significant increase of the peak current for methylene blue upon the hybridization of immobilized ssDNA with cDNA in the solution was observed. This peak current change was used to monitor the recognition of CML DNA sequence. This electrochemical approach is sequence specific as indicated by the control experiments in which no peak current change was observed if a non-complementary DNA sequence was used. Factors, such as DNA target concentration and hybridization conditions determining the sensitivity of the electrochemical assay were investigated. Under optimal conditions, this sensor has a good calibration range between 1.25 x 10(-7) and 6.75 x 10(-7) M, with CML DNA sequence detection limit of 5.9 x 10(-8) M. (c) 2006 Elsevier B.V. All rights reserved.
引用
收藏
页码:468 / 471
页数:4
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