Desenvolvimento de um aptassensor para detecção do vírus da dengue
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Data
2017-08-17
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Universidade Federal de Goiás
Resumo
Dengue is an endemic disease that causes wide concern both the health systems and to the patients. Thus, it is important that studies aiming at diagnoses faster and with possibility of field application can be developed for the medical intervention happens early, avoiding the worsening of the cases. Therefore, the aim of this work was to develop an electrochemical biosensor based on the use of aptamer as a bioelement for recognition on the surface of the boron doped diamond electrode (BDD) for the detection of each of the four serotypes of the dengue virus. Previously to the development of the biosensor, the dot-blot and apta-PCR techniques were used as validation methods for the interaction between aptamer B07 and target, the 5'UTR region present in the viral genome. The cyclic voltammetry technique was used in the analysis of redox reactions on the surface of the BDD electrode with and without modification of its surface with chitosan film. Ferrocene was used as the redox mediator and electroactive indicator of hybridization of the DNA strands formed in the sensor. The peaks of current indicated that the electrode with chitosan film modification on its surface presented greater stability. The redox compound had higher affinity for the double chains hybridized on the surface of the electrode, showing current values for DENV 1, 2, 3 and 4 of were 0.73; 0.69; 0.79 and 1.03 μA, respectively. These values were higher than the current found for the single-stranded aptamer (ssDNA), which was 0.62 μA, as well as for the current obtained from the aptamer hybridized with its complementary strand (dsDNA) whose value was 0.91 μA. Analyzes with time variations were performed showing a reduction in current values as a function of time, probably due to the reduction of the interaction of the electroactive material in the sensor. The aptasensor developed here showed good detection distinction between nucleic acid sequences, presenting potential for application in the detection of dengue virus.
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Palavras-chave
Aptâmero, Dengue, Ferroceno, Detecção eletroquímica, Voltametria cíclica, Quitosana, Aptamer, Ferrocene, Electrochemical detection, Cyclic voltammetry, Chitosan
Citação
SOUZA, Evellyn Gonçalves de. Desenvolvimento de um aptassensor para detecção do vírus da dengue. 2017. 56 f. Dissertação (Mestrado em Química) - Universidade Federal de Goiás, Catalão, 2017.