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Electrochemical Biosensor for Sensitive Detection of Hepatitis B in Human Plasma

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Abstract

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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Data Availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

The authors are grateful for the financial support from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).

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Authors and Affiliations

  1. Institute of Biotechnology, Federal University of Uberlândia, Uberlândia, Brazil

    Ana Cristina Honorato de Castro, José Manuel Rodrigueiro Flauzino & Ana Graci Brito-Madurro

  2. Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil

    Leandro Toshio Kochi

  3. Adolfo Lutz Institute, Regional Laboratory in, São José Do Rio Preto, Brazil

    Márcia Maria Costa Nunes Soares

  4. Chemistry Department, Federal University of Triângulo Mineiro, Uberaba, Brazil

    Valéria Almeida Alves & Luís Antônio da Silva

  5. Institute of Chemistry, Federal University of Uberlândia, Uberlândia, Brazil

    João Marcos Madurro

Authors
  1. Ana Cristina Honorato de Castro
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  2. Leandro Toshio Kochi
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  3. José Manuel Rodrigueiro Flauzino
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  4. Márcia Maria Costa Nunes Soares
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  5. Valéria Almeida Alves
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  6. Luís Antônio da Silva
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  7. João Marcos Madurro
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  8. Ana Graci Brito-Madurro
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Contributions

Ana C. H. Castro: conceptualization, methodology, formal analysis, investigation, writing. Leandro T. Kochi: methodology, investigation, conceptualization, writing. José M. R. Flauzino: methodology, investigation, validation, formal analysis. Márcia M. C. N. Soares: human serum sample supply, methodology. Valéria A. Alves: methodology, formal analysis. Luís A. da Silva: methodology, formal analysis. Joao M. Madurro: investigation, conceptualization, supervision, writing – reviewing and editing, funding acquisition. Ana G. Brito Madurro: investigation, conceptualization, supervision, writing – reviewing and editing, funding acquisition.

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Correspondence to Ana Graci Brito-Madurro.

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de Castro, A.C.H., Kochi, L.T., Flauzino, J.M.R. et al. Electrochemical Biosensor for Sensitive Detection of Hepatitis B in Human Plasma. Appl Biochem Biotechnol 194, 2604–2619 (2022). https://doi.org/10.1007/s12010-022-03829-4

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