Development of a New Route for the Immobilization of Unmodified Single-Stranded DNA on Chitosan Beads and Detection of Released Guanine after Hydrolysis

被引：11

作者：

Chahri, Ikram ^{[1
]}

Karrat, Abdelhafid ^{[1
]}

Mohammadi, Hasna ^{[1
]}

Amine, Aziz ^{[1
]}

机构：

[1] Hassan II Univ Casablanca, Fac Sci & Tech, Lab Proc Engn & Environm, PA146, Mohammadia 28810, Morocco

来源：

MOLECULES | 2023年 / 28卷 / 05期

关键词：

DNA immobilization; guanine molecules; hydrolysis; differential pulse voltammetry; chitosan beads; electrochemical biosensors; ELECTROCHEMICAL BIOSENSING PLATFORM; GRAPHITE ELECTRODE; AU NANOPARTICLES; CARBON-BLACK; AMPLIFICATION; HYBRIDIZATION; SOLUBILITY; SENSOR; WATER; FILM;

D O I：

10.3390/molecules28052088

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

In this work, chitosan beads were used as a cost-effective platform for the covalent immobilization of unmodified single-stranded DNA, using glutaraldehyde as a cross-linking agent. The immobilized DNA capture probe was hybridized in the presence of miRNA-222 as a complementary sequence. The target was evaluated based on the electrochemical response of the released guanine, using hydrochloride acid as a hydrolysis agent. Differential pulse voltammetry technique and screen-printed electrodes modified with COOH-functionalized carbon black were used to monitor the released guanine response before and after hybridization. The functionalized carbon black provided an important signal amplification of guanine compared to the other studied nanomaterials. Under optimal conditions (6 M HCl at 65 degrees C for 90 min), an electrochemical-based label-free genosensor assay exhibited a linear range between 1 nM and 1 mu M of miRNA-222, with a detection limit of 0.2 nM of miRNA-222. The developed sensor was successfully used to quantify miRNA-222 in a human serum sample.

引用

页数：13

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