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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 peaks related to guanine oxidation of the probe-modified-PGE after (a) and before hybridization with DENV-3 (b); in the presence of non-complementary sequences (c) and in a solution mixed with DENV-3 and non-complementary sequences (d), all diluted in human serum. 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-f006: Current peaks related to guanine oxidation of the probe-modified-PGE after (a) and before hybridization with DENV-3 (b); in the presence of non-complementary sequences (c) and in a solution mixed with DENV-3 and non-complementary sequences (d), all diluted in human serum. Experimental conditions: Scanning potential range between +0.5 V and +1.2 V and scan rate of 0.05 V/s.

Mentions: As shown in Figure 6, the biosensor displays the same electrochemical behavior observed previously in Tris-HCl buffer solutions. However, all the current signals of the probe-modified PGE after hybridization with the complementary, non-complementary sequences and mixed DNA solution diluted in human serum presented a slight decrease (134, 410.8 and 221 nA, respectively) when compared with those diluted in acetate buffer (135, 600 and 230 nA, respectively). This could be due to the hybridization kinetics and the efficiency of the PGE surface, which could be affected by non-specific adsorption of plasma proteins, and this may interfere with the detection of the electrochemical signal [63]. However, such interference with the detection of DNA molecules was observed previously with the optical DNA biosensor developed by Gong et al. [64,65]. Thus, these results confirm the high selectivity and sensitivity of the electrochemical DNA biosensor developed herein.


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 peaks related to guanine oxidation of the probe-modified-PGE after (a) and before hybridization with DENV-3 (b); in the presence of non-complementary sequences (c) and in a solution mixed with DENV-3 and non-complementary sequences (d), all diluted in human serum. 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

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

sensors-15-15562-f006: Current peaks related to guanine oxidation of the probe-modified-PGE after (a) and before hybridization with DENV-3 (b); in the presence of non-complementary sequences (c) and in a solution mixed with DENV-3 and non-complementary sequences (d), all diluted in human serum. Experimental conditions: Scanning potential range between +0.5 V and +1.2 V and scan rate of 0.05 V/s.
Mentions: As shown in Figure 6, the biosensor displays the same electrochemical behavior observed previously in Tris-HCl buffer solutions. However, all the current signals of the probe-modified PGE after hybridization with the complementary, non-complementary sequences and mixed DNA solution diluted in human serum presented a slight decrease (134, 410.8 and 221 nA, respectively) when compared with those diluted in acetate buffer (135, 600 and 230 nA, respectively). This could be due to the hybridization kinetics and the efficiency of the PGE surface, which could be affected by non-specific adsorption of plasma proteins, and this may interfere with the detection of the electrochemical signal [63]. However, such interference with the detection of DNA molecules was observed previously with the optical DNA biosensor developed by Gong et al. [64,65]. Thus, these results confirm the high selectivity and sensitivity of the electrochemical DNA biosensor developed herein.

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