Direct detection and discrimination of double-stranded oligonucleotide corresponding to hepatitis C virus genotype 3a using an electrochemical DNA biosensor based on peptide nucleic acid and double-stranded DNA hybridization

被引:47
作者
Pournaghi-Azar, M. H. [1 ]
Ahour, F. [1 ]
Hejazi, M. S. [2 ,3 ,4 ]
机构
[1] Univ Tabriz, Electroanalyt Chem Lab, Fac Chem, Tabriz 5166614776, Iran
[2] Tabriz Univ Med Sci, Fac Pharm, Tabriz 51664, Iran
[3] Tabriz Univ Med Sci, Drug Appl Res Ctr, Tabriz 51664, Iran
[4] Tabriz Univ Med Sci, Pharmaceut Nanotechnol Res Ctr, Tabriz 51664, Iran
关键词
DNA biosensor; Hepatitis C; PNA probe; Triplex formation; Gold electrode; PENCIL GRAPHITE ELECTRODE; ION-CHANNEL SENSORS; HUMAN INTERLEUKINE-2 GENE; LABEL-FREE; VOLTAMMETRIC DETECTION; ARTIFICIAL RECEPTORS; POINT MUTATION; GOLD SURFACES; PNA; PROBE;
D O I
10.1007/s00216-010-3875-5
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Development of an electrochemical DNA biosensor for the direct detection and discrimination of double-stranded oligonucleotide ( dsDNA) corresponding to hepatitis C virus genotype 3a, without its denaturation, using a gold electrode is described. The electrochemical DNA sensor relies on the modification of the gold electrode with 6-mercapto-1-hexanol and a self-assembled monolayer of 14-mer peptide nucleic acid probe, related to the hepatitis C virus genotype 3a core/E1 region. The increase of differential pulse voltammetric responses of methylene blue, upon hybridization of the self-assembled probe with the target ds-DNA to form a triplex is the principle behind the detection and discrimination. Some hybridization experiments with non-complementary oligonucleotides were carried out to assess whether the developed DNA sensor responds selectively to the ds-DNA target. Diagnostic performance of the biosensor is described and the detection limit was found to be 1.8x10(-12) M in phosphate buffer solution, pH 7.0. The relative standard deviation of measurements of 100 pM of target ds-DNA performed with three independent probe-modified electrodes was 3.1%, indicating a remarkable reproducibility of the detection method.
引用
收藏
页码:3581 / 3587
页数:7
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