Rapid, Sensitive, and Label-Free Impedimetric Detection of a Single-Nucleotide Polymorphism Correlated to Kidney Disease

被引:17
作者
Bonanni, Alessandra [1 ]
Pumera, Martin [1 ]
Miyahara, Yuji [1 ]
机构
[1] NIMS, Ctr Biomat, Int Ctr Mat Nanoarchitecton MANA, Ibaraki, Japan
关键词
ELECTROCHEMICAL IMPEDANCE SPECTROSCOPY; DNA HYBRIDIZATION DETECTION; COL4A5; GENE; SENSOR; ELECTRODES; BIOSENSORS; POLYMER; DIFFERENTIATION; TRANSDUCTION; VOLTAMMETRY;
D O I
10.1021/ac100165q
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
We present a protocol for the very rapid and sensitive detection of a specific mutation of the COL4A5 gene (exon 29, A-C mismatch) which was found in people affected by Alport syndrome (AS) and their families. Disposable electrochemically printed electrodes were used to immobilize a single-stranded oligonucleotide probe that was complementary to the AS-correlated gene. The detection principle is based on changes in the impedance spectra of the redox probe ferro/ferricyanide after hybridization with synthetic target DNA. Detection was performed either for mutated or for healthy (wild-type) gene copies. The high sensitivity obtained with this protocol (LOD in the picomolar range) was additionally enhanced to the femtomolar range by performing the detection in the presence of Ca2+. In fact, the specific binding of the metal ions in the presence of an A-C nucleotide mismatch induced a further impedance change, thus improving the discrimination between the mutated and healthy gene, as the signal amplification is achieved only for the former.
引用
收藏
页码:3772 / 3779
页数:8
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