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Electroanalytical Paper-Based Nucleic Acid Amplification Biosensors with Integrated Thread Electrodes.

Shirin Khaliliazar1, Anna Toldrà1, Georgios Chondrogiannis1

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This study introduces a 3D microfluidic paper-based electrochemical nucleic acid amplification test (NAAT) for rapid point-of-care diagnostics. The novel device enables sensitive detection of toxic microalgae DNA in under 2 hours.

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Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Environmental Science

Background:

  • Nucleic acid amplification tests (NAATs) are highly sensitive and specific but typically require centralized labs, limiting their use in point-of-care settings.
  • Developing portable and rapid diagnostic tools is crucial for timely detection of biological threats and environmental monitoring.

Purpose of the Study:

  • To develop a 3D microfluidic paper-based electrochemical NAAT for point-of-care applications.
  • To demonstrate the device's capability for detecting the toxic microalgae *Ostreopsis* cf. *ovata*.

Main Methods:

  • Integration of gold plasma-coated threads for electroanalytical readouts within a 3D microfluidic paper device.
  • Utilizing movable filter paper stacks for sequential assay steps including incubation, rinsing, and detection.
  • Employing glass fiber substrates for reagent storage and isothermal amplification via recombinase polymerase amplification.

Main Results:

  • The developed NAAT achieved a limit of detection of 0.06 pM for synthetic DNA and detected 1 ng/μL of *O. *cf. *ovata* genomic DNA.
  • The entire assay was completed within 95 minutes with negligible cross-reactivity.
  • Demonstrated successful detection of a specific toxic microalgae species.

Conclusions:

  • The integration of thread electrodes in paper-based devices offers a promising platform for developing digital, single-use NAATs.
  • This technology has the potential for various advanced electroanalytical applications in paper- or textile-based formats.