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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

Electrochemical signals of different concentrations of DENV-3 probe onto pencil graphite electrodes (PGE). Differential pulse voltammetry (DPV) was used for electrochemical analysis based on guanine oxidation. Experimental conditions: Scanning potential range between +0.5 V and +1.2 V and scan rate of 0.05 V/s. The results represent the average of triplicates carried out at each DENV-3 probe concentration.
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sensors-15-15562-f002: Electrochemical signals of different concentrations of DENV-3 probe onto pencil graphite electrodes (PGE). Differential pulse voltammetry (DPV) was used for electrochemical analysis based on guanine oxidation. Experimental conditions: Scanning potential range between +0.5 V and +1.2 V and scan rate of 0.05 V/s. The results represent the average of triplicates carried out at each DENV-3 probe concentration.

Mentions: Figure 2 shows current peaks of different DENV-3 probe concentrations on the PGE surface. As the electrochemical analysis in this study relies on label-free oxidation of guanine bases, the acquisition of higher current signals for DNA probes is well-suited for this system [29,48,49,50]. The results show that the current gradually rises with the increase of the probe concentration from 250 nM up to 500 nM, reaching the highest electrochemical signal of 777 ± 8.6 nA at 500 nM. The result obtained at 500 nM was also statistically different from that obtained at 750 nM (p = 0.000178). However, the decrease in the current peaks at higher concentrations of DNA probes after 500 nM could be due to the steric hindrance between the nitrogenous bases and the transducer. This prevents the electrons produced by the oxidation process to access the electrode surface [51,52,53,54,55]. Therefore, a concentration of 500 nM was selected as the optimal probe concentration for DNA immobilization on the PGE.


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)

Electrochemical signals of different concentrations of DENV-3 probe onto pencil graphite electrodes (PGE). Differential pulse voltammetry (DPV) was used for electrochemical analysis based on guanine oxidation. Experimental conditions: Scanning potential range between +0.5 V and +1.2 V and scan rate of 0.05 V/s. The results represent the average of triplicates carried out at each DENV-3 probe concentration.
© Copyright Policy
Related In: Results  -  Collection

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

sensors-15-15562-f002: Electrochemical signals of different concentrations of DENV-3 probe onto pencil graphite electrodes (PGE). Differential pulse voltammetry (DPV) was used for electrochemical analysis based on guanine oxidation. Experimental conditions: Scanning potential range between +0.5 V and +1.2 V and scan rate of 0.05 V/s. The results represent the average of triplicates carried out at each DENV-3 probe concentration.
Mentions: Figure 2 shows current peaks of different DENV-3 probe concentrations on the PGE surface. As the electrochemical analysis in this study relies on label-free oxidation of guanine bases, the acquisition of higher current signals for DNA probes is well-suited for this system [29,48,49,50]. The results show that the current gradually rises with the increase of the probe concentration from 250 nM up to 500 nM, reaching the highest electrochemical signal of 777 ± 8.6 nA at 500 nM. The result obtained at 500 nM was also statistically different from that obtained at 750 nM (p = 0.000178). However, the decrease in the current peaks at higher concentrations of DNA probes after 500 nM could be due to the steric hindrance between the nitrogenous bases and the transducer. This prevents the electrons produced by the oxidation process to access the electrode surface [51,52,53,54,55]. Therefore, a concentration of 500 nM was selected as the optimal probe concentration for DNA immobilization on the PGE.

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