New approach to immobilization and specific-sequence detection of nucleic acids based on poly(4-hydroxyphenylacetic acid)

Immobilization and hybridization of oligonucleotides or specific-gene PCR product (DENV-1), a conserved genomic sequence of the dengue virus, onto graphite electrode modified with poly(4-hydroxyphenylacetic acid), were carried out with success using both direct electrochemical oxidation of guanine or redox electroactive indicator ethidium bromide.Studies of oligonucleotides hybridization with the complementary target showed a decrease of both guanosine and adenosine current peaks, when compared with the peak previously obtained before the hybridization. Immobilized ssDNA, DENV-1, was hybridized with various concentrations of target DNA. The interaction between DENV-1 hybridized onto the modified graphite electrodes surface and the intercalator, ethidium bromide, was observed by differential pulse voltammetry, monitoring the current change generated to the DNA intercalator accumulated onto the modified electrode after DNA hybridization. For the determination of complementary target, the proposed method exhibited a good dynamic range (12–42 nmol L^? 1) and a low detection limit (7.12 nmol L^? 1).AFM images showed that the oligonucleotides or single-stranded DNA, DENV-1, before hybridization, had roughness values lower than the double stranded obtained after hybridization.The new surface obtained in these work, as well as the possibility of utilization of the same to monitor hybridization events is a promising strategy for the development of DNA electrochemical biosensors.