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A Sensitive and Selective Label-Free Electrochemical DNA Biosensor for the Detection of Specific Dengue Virus Serotype 3 Sequences.

Oliveira N, Souza E, Ferreira D, Zanforlin D, Bezerra W, Borba MA, Arruda M, Lopes K, Nascimento G, Martins D, Cordeiro M, Lima-Filho J - Sensors (Basel) (2015)

Bottom Line: 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).Moreover, this system allows selective detection of DENV-3 sequences in buffer and human serum solutions.

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Imunopatologia Keizo Asami (LIKA), Universidade Federal de Pernambuco-UFPE, Av. Prof. Moraes Rego, s/n, Campus da UFPE, 50670-901 Recife, PE, Brazil. nataliacybelle.89@gmail.com.

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.

No MeSH data available.


Related in: MedlinePlus

Current signals obtained for different DENV-3 target sequence concentrations after hybridization with probe-modified PGEs. Inset: Related calibration graph at a concentration range of 10–100 nM for the target sequence. Experimental conditions: Scanning potential range between +0.5 V and +1.2 V and scan rate of 0.05 V/s.
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sensors-15-15562-f003: Current signals obtained for different DENV-3 target sequence concentrations after hybridization with probe-modified PGEs. Inset: Related calibration graph at a concentration range of 10–100 nM for the target sequence. Experimental conditions: Scanning potential range between +0.5 V and +1.2 V and scan rate of 0.05 V/s.

Mentions: In this study, the biosensor performance was analyzed through the hybridization reaction between the DENV-3 probe and the complementary DENV-3 oligonucleotide. Hybridization was carried out with different amounts of the target sequence and this reaction was performed in an electrochemical cell containing 20 mM Tris-HCl buffer (pH 7.0). The electrochemical signals based on guanine oxidation are displayed in Figure 3. The results showed that the current peaks increase with the increasing concentration of the target sequence (10 nM to 500 nM); the highest concentration exhibited the highest current peak of the system (135 ± 2.15 nA). However, at concentrations higher than 500 nM, there is a decrease in the electrochemical signal that could be due to electrostatic hindrance of DNA molecules on the PGE surface [39,53,56].


A Sensitive and Selective Label-Free Electrochemical DNA Biosensor for the Detection of Specific Dengue Virus Serotype 3 Sequences.

Oliveira N, Souza E, Ferreira D, Zanforlin D, Bezerra W, Borba MA, Arruda M, Lopes K, Nascimento G, Martins D, Cordeiro M, Lima-Filho J - Sensors (Basel) (2015)

Current signals obtained for different DENV-3 target sequence concentrations after hybridization with probe-modified PGEs. Inset: Related calibration graph at a concentration range of 10–100 nM for the target sequence. Experimental conditions: Scanning potential range between +0.5 V and +1.2 V and scan rate of 0.05 V/s.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4541844&req=5

sensors-15-15562-f003: Current signals obtained for different DENV-3 target sequence concentrations after hybridization with probe-modified PGEs. Inset: Related calibration graph at a concentration range of 10–100 nM for the target sequence. Experimental conditions: Scanning potential range between +0.5 V and +1.2 V and scan rate of 0.05 V/s.
Mentions: In this study, the biosensor performance was analyzed through the hybridization reaction between the DENV-3 probe and the complementary DENV-3 oligonucleotide. Hybridization was carried out with different amounts of the target sequence and this reaction was performed in an electrochemical cell containing 20 mM Tris-HCl buffer (pH 7.0). The electrochemical signals based on guanine oxidation are displayed in Figure 3. The results showed that the current peaks increase with the increasing concentration of the target sequence (10 nM to 500 nM); the highest concentration exhibited the highest current peak of the system (135 ± 2.15 nA). However, at concentrations higher than 500 nM, there is a decrease in the electrochemical signal that could be due to electrostatic hindrance of DNA molecules on the PGE surface [39,53,56].

Bottom Line: 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).Moreover, this system allows selective detection of DENV-3 sequences in buffer and human serum solutions.

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Imunopatologia Keizo Asami (LIKA), Universidade Federal de Pernambuco-UFPE, Av. Prof. Moraes Rego, s/n, Campus da UFPE, 50670-901 Recife, PE, Brazil. nataliacybelle.89@gmail.com.

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.

No MeSH data available.


Related in: MedlinePlus