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DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
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Direct DNA Analysis with Paper-Based Ion Concentration Polarization.

Max M Gong1, Reza Nosrati1, Maria C San Gabriel2

  • 1Department of Mechanical and Industrial Engineering, University of Toronto , 5 King's College Rd., Toronto, Ontario, Canada M5S 3G8.

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Direct DNA analysis is now possible on paper-based devices using ion concentration polarization. This technology enables rapid, sensitive detection of Hepatitis B virus DNA and accurate assessment of sperm DNA integrity, improving diagnostics accessibility.

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Area of Science:

  • Biotechnology
  • Molecular Diagnostics
  • Nanotechnology

Background:

  • Conventional DNA testing requires laboratory settings, limiting accessibility.
  • Point-of-care diagnostics are crucial for improving patient care globally.
  • Novel technologies are needed to bring advanced molecular diagnostics closer to patients.

Purpose of the Study:

  • To demonstrate direct DNA analysis using paper-based devices.
  • To enable rapid preconcentration, separation, and detection of DNA targets.
  • To assess the utility of paper-based ion concentration polarization (ICP) for disease diagnosis and fertility testing.

Main Methods:

  • Development of paper-based devices utilizing ion concentration polarization (ICP) at nanoporous membranes.
  • Simultaneous preconcentration, separation, and detection of Hepatitis B virus DNA in human serum.
  • Clinical assessment of sperm DNA fragmentation using paper-based ICP devices and comparison with the sperm chromatin structure assay.

Main Results:

  • Achieved a limit of detection of 150 copies/mL for Hepatitis B virus DNA without amplification, enabling early diagnosis.
  • Demonstrated 100% agreement with clinical decisions for sperm DNA fragmentation assessment.
  • Showcased a strong correlation (R² = 0.98) between paper-based ICP results and the sperm chromatin structure assay.

Conclusions:

  • Paper-based ICP technology facilitates direct, rapid, and sensitive DNA analysis.
  • This approach offers inexpensive and accessible advanced molecular diagnostics.
  • The technology has significant potential for improving disease diagnosis and fertility assessments in diverse settings.