Electrochemical Biosensor for Sensitive Detection of Hepatitis B in Human Plasma
2022
de Castro, Ana Cristina Honorato | Kochi, Leandro Toshio | Flauzino, José Manuel Rodrigueiro | Soares, Márcia Maria Costa Nunes | Alves, Valéria Almeida | da Silva, Luís Antônio | Madurro, João Marcos | Brito-Madurro, Ana Graci
In this work, we report the construction of a novel electrochemical device for molecular diagnosis of hepatitis B virus in the blood plasma of infected patients, using graphite electrodes functionalized with poly(4-aminophenol) and sensitized with a specific DNA probe. The recognition of genomic DNA was evaluated by electrochemical techniques (DPV and EIS) and scanning electron microscopy. The genosensor was efficient in detecting genomic DNA with a linear range from 1.176 to 4.825 μg mL⁻¹ and detection limit of 35.69 ng mL⁻¹ (4.63 IU ml⁻¹ or 25.93 copies.ml⁻¹), which is better than the 10.00 IU ml⁻¹ limit of reference method, real-time PCR, used in point of care. EIS analysis shows that the genosensor resistance increased exponentially with the concentration of the genomic DNA target. This novel platform has advantages to its applicability in real samples, such as good sensitivity, selectivity, low sample volume, and fast assay time (36 min), thus interesting for application in the diagnosis of hepatitis B virus in blood plasma. Also, the ease of synthesis of the low-cost polymer by electrosynthesis directly on the electrode surface allows the translation of the platform to portable devices.
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