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“Do it yourself” protocol to fabricate dual-detection paper-based analytical device for salivary biomarker analysis

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Abstract

This paper describes the design and construction of dual microfluidic paper–based analytical devices (dual-μPADs) as a lab-on-paper platform involving a “do-it-yourself” fabrication protocol. The device comprises a colorimetric and electrochemical module to obtain a dual-mode signal readout sensing strategy. A 3D pen polymeric resin was used to prepare graphite carbon-based electrodes and hydrophobic barriers on paper substrates. The proposed carbon-based ink was employed to manufacture electrodes on paper based on a stencil-printing approach, which were further characterized by electrochemical and morphological analyses. The analytical performance of the dual-μPADs was simultaneously evaluated for lactate, pH, nitrite, and salivary amylase (sAA) analysis. To demonstrate the proof-of-concept, saliva samples collected from both healthy individuals and those with periodontitis were successfully tested to demonstrate the feasibility of the proposed devices. Samples collected from individuals previously diagnosed with periodontitis showed high levels of nitrite and sAA (> 94 μmol L−1 and > 610 U mL−1) in comparison with healthy individuals (≤ 16 μmol L−1 and 545 U mL−1). Moreover, periodontitis saliva resulted in acid solution and almost null lactate levels. Notably, this protocol supplies a simple way to manufacture dual-μPADs, a versatile platform for sensitive detecting of biomarkers in saliva playing a crucial role towards the point-of-care diagnosis of periodontal disease.

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Funding

The authors would like to thank CNP (grant 307554/2020–1 and 405620/2021–7), CAPES (grant 88887.192880/2018–00 and finance code 001), FAPEG and INCTBio (grant 465389/2014–7) for the financial support and granted scholarships. CONICET and CNPq are thanked for the scholarships granted to H.A.S.N., and L.F.C. The authors also acknowledge the High-Resolution Multi-User Microscopy Laboratory (LabMic) from Federal University of Goias for providing SEM analysis and the School of Dentistry from Federal University of Goiás for supporting the collection of saliva samples.

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Authors and Affiliations

  1. Instituto de Química, Universidade Federal de Goiás, Campus Samambaia, Goiânia, GO, 74690-900, Brazil

    Lucas R. Sousa, Habdias A. Silva-Neto, Lucas F. Castro, Karoliny A. Oliveira & Wendell K. T. Coltro

  2. Laboratorio de Biosensores y Bioanálisis (LABB), Departamento de Química Biológica e IQUIBICEN−CONICET, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires (UBA), Pabellón 2, Ciudad Universitaria, Ciudad Autónoma de Buenos Aires, Argentina

    Lucas R. Sousa, Federico Figueredo & Eduardo Cortón

  3. Instituto Nacional de Ciência e Tecnologia de Bioanalítica, Campinas, SP, 13084-971, Brazil

    Wendell K. T. Coltro

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  1. Lucas R. Sousa
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  2. Habdias A. Silva-Neto
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Correspondence to Wendell K. T. Coltro.

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Saliva samples were collected in accordance with the ethical principles involving human participants under the approval of the ethical committee from Federal University of Goias (CAAE n° 68271817.9.0000.5083).

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Published in the topical collection Young Investigators in (Bio-)Analytical Chemistry 2023 with guest editors Zhi-Yuan Gu, Beatriz Jurado-Sánchez, Thomas H. Linz, Leandro Wang Hantao, Nongnoot Wongkaew, and Peng Wu.

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Sousa, L.R., Silva-Neto, H.A., Castro, L.F. et al. “Do it yourself” protocol to fabricate dual-detection paper-based analytical device for salivary biomarker analysis. Anal Bioanal Chem 415, 4391–4400 (2023). https://doi.org/10.1007/s00216-023-04581-2

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