Abstract

Dengue fever is the most prevalent vector-borne disease in the world, with nearly 100 million people infected every year. Early diagnosis and identification of the pathogen are crucial steps for the treatment and for prevention of the disease, mainly in areas where the co-circulation of different serotypes is common, increasing the outcome of dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Due to the lack of fast and inexpensive methods available for the identification of dengue serotypes, herein we report the development of an electrochemical DNA biosensor for the detection of sequences of dengue virus serotype 3 (DENV-3). DENV-3 probe was designed using bioinformatics software and differential pulse voltammetry (DPV) was used for electrochemical analysis. The results showed that a 22-m sequence was the best DNA probe for the identification of DENV-3. The optimum concentration of the DNA probe immobilized onto the electrode surface is 500 nM and a low detection limit of the system (3.09 nM). Moreover, this system allows selective detection of DENV-3 sequences in buffer and human serum solutions. Therefore, the application of DNA biosensors for diagnostics at the molecular level may contribute to future advances in the implementation of specific, effective and rapid detection methods for the diagnosis dengue viruses.

Highlights

  • A Sensitive and Selective Label-Free Electrochemical DNANatália Oliveira 1,*, Elaine Souza 2, Danielly Ferreira 1, Deborah Zanforlin 1, Wessulla Bezerra 1, Maria Amélia Borba 1, Mariana Arruda 1, Kennya Lopes 3, Gustavo Nascimento 1, Danyelly Martins 1,4, Marli Cordeiro 4 and José Lima-Filho 1,4

  • Dengue fever is the most prevalent vector-borne disease in the world

  • This can be achieved using bioinformatics analysis based on whole genome sequencing, in a way to predict the most specific region that is able to produce a steady double-strand DNA with the pathogen [37,38]

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Summary

A Sensitive and Selective Label-Free Electrochemical DNA

Natália Oliveira 1,*, Elaine Souza 2, Danielly Ferreira 1, Deborah Zanforlin 1, Wessulla Bezerra 1, Maria Amélia Borba 1, Mariana Arruda 1, Kennya Lopes 3, Gustavo Nascimento 1, Danyelly Martins 1,4, Marli Cordeiro 4 and José Lima-Filho 1,4.

Introduction
Design of a Specific DENV-3 DNA Probe
Reagents and Materials
Apparatus
Preparation of Electrodes and Pre-Treatment of PGE
DNA Probe Immobilization onto PGE Surface
DNA Hybridization with Complementary and Non-Complementary Sequences
Detection of Complementary and Non-Complementary Sequences in Human Serum
Electrochemical Analysis
Statistical Data Analysis
Bioinformatics Analysis of DENV-3 DNA Probes
Effect of DENV-3 Probes Concentration on Immobilization on the PGE
Electrochemical Analysis of Hybridization Assays
Selectivity Study
Electrochemical Measurement of Target Hybridization in Human Serum Solutions
Conclusions
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