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In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
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Kinetic ELISA in microfluidic channels.

Naoki Yanagisawa1, Debashis Dutta2

  • 1Department of Chemistry, University of Wyoming, Laramie, WY 82071, USA. nyanagis@uwyo.edu.

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|January 15, 2015
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Summary

This study introduces a kinetic ELISA in microfluidic channels for detecting Blue Tongue and Epizootic Hemorrhagic Disease viral antibodies. This method offers significantly lower detection limits and faster results compared to traditional assays.

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Area of Science:

  • Veterinary Virology
  • Analytical Chemistry
  • Biotechnology

Background:

  • Traditional enzyme-linked immunosorbent assays (ELISA) for viral antibody detection can be time-consuming and require larger sample volumes.
  • Microfluidic platforms offer potential for miniaturization and improved assay performance.
  • Kinetic ELISA formats can enhance sensitivity by measuring reaction rates rather than end-point signals.

Purpose of the Study:

  • To develop and validate a kinetic ELISA for quantifying Blue Tongue and Epizootic Hemorrhagic Disease viral antibodies in microfluidic channels.
  • To compare the performance of this microfluidic kinetic ELISA with conventional end-point ELISA on microwell plates.

Main Methods:

  • Utilized microfluidic channels fabricated on inexpensive glass microchips.
  • Employed a kinetic ELISA approach by monitoring enzyme reaction product generation rate under static conditions.
  • Quantified viral antibodies using an epifluorescence microscope coupled to a CCD camera.

Main Results:

  • Achieved a limit of detection (LOD) 3-5 times lower than commercial microwell plates, using 1/50th the sample volume and less than 1/3rd the incubation time.
  • Demonstrated an improvement in LOD by over an order of magnitude compared to end-point ELISA in microfluidic devices.
  • Observed substantially lower noise levels in kinetic ELISA due to reliance on signal generation rate, unlike end-point ELISA which relies on absolute fluorescence measurements.

Conclusions:

  • Kinetic ELISA in microfluidic channels provides a highly reliable and sensitive method for viral antibody quantitation.
  • This approach offers significant advantages in terms of sensitivity, speed, and sample volume compared to traditional ELISA formats.
  • Microfluidic kinetic ELISA represents a promising advancement for the rapid and accurate diagnosis of viral diseases.