Development of a quantitative thyroid-stimulating hormone assay system for a benchtop digital ELISA desktop analyzer

Area of Science:

• Biochemistry
• Immunoassay Technology
• Clinical Diagnostics

Background:

• Thyroid-stimulating hormone (TSH) testing is crucial for diagnosing thyroid disorders.
• Traditional enzyme-linked immunosorbent assay (ELISA) is a standard method for TSH detection.
• Next-generation digital immunoassays (d-IA) offer enhanced molecular detection sensitivity.

Purpose of the Study:

• To develop and evaluate a TSH assay system utilizing the digital immunoassay (d-IA) platform.
• To assess the performance of the d-IA TSH assay with small sample volumes.
• To compare the functional sensitivity of the d-IA TSH assay with existing methods.

Main Methods:

• Development of a TSH assay on the d-IA platform.
• Assay validation using minimal sample volumes (5 µL).
• Determination of limits of blank, detection, and quantification (functional sensitivity).
• Evaluation of assay precision and correlation between serum and plasma samples.

Main Results:

• The d-IA TSH assay successfully measured TSH with a sample volume as low as 5 µL.
• Achieved limits of blank, detection, and quantification of 0.000346, 0.001953, and 0.002280 μIU/mL, respectively.
• Demonstrated high precision (total coefficient of variation ≤ 10%) and good agreement between serum and plasma measurements.
• Functional sensitivity (0.002280 μIU/mL) was comparable to third-generation assays (e.g., ARCHITECT TSH assay at 0.0038 μIU/mL).

Conclusions:

• The developed d-IA TSH assay is a sensitive and efficient method for TSH measurement.
• The assay's ability to use small sample volumes makes it suitable for various clinical applications.
• This technology provides a valuable tool for thyroid disease diagnosis with high accuracy and sensitivity.