Proposal of a Rapid Detection System Using Image Analysis for ELISA with an Autonomous Centrifugal Microfluidic...

Area of Science:

• Biomedical Engineering
• Analytical Chemistry
• Diagnostics

Background:

• Enzyme-linked immunosorbent assays (ELISA) are widely used but often time-consuming.
• Adapting ELISA for point-of-care testing (POCT) requires significant reduction in assay duration.
• Conventional ELISA methods involve multiple steps and often require reaction stopping reagents.

Purpose of the Study:

• To develop and validate an image analysis method for ELISA on a microfluidic device.
• To adapt ELISA for rapid point-of-care testing (POCT) by reducing assay time.
• To eliminate the need for reaction stopping reagents in ELISA.

Main Methods:

• Utilized a centrifugal microfluidic device for automated ELISA execution.
• Developed an image analysis technique to quantify color development on a 3,3',5,5'-tetramethylbenzidine (TMB) substrate.
• Performed assay and image analysis without a reaction stopping reagent.

Main Results:

• Successfully prepared a calibration curve for mouse IgG detection.
• Demonstrated that the image analysis method achieved performance comparable to conventional methods.
• Obtained a calibration curve within 2 minutes of TMB substrate addition, with a total assay time of 10 minutes.

Conclusions:

• The developed image analysis method for ELISA on microfluidic devices significantly reduces assay time to 10 minutes.
• This rapid ELISA system is suitable for point-of-care testing (POCT) applications.
• The method eliminates the need for reaction stopping reagents, simplifying the assay process.