Paper-based chemiluminescence enzyme-linked immunosorbent assay enhanced by biotin-streptavidin system for...

Area of Science:

• Biomedical Engineering
• Analytical Chemistry
• Cardiovascular Diagnostics

Background:

• High-sensitivity C-reactive protein (hs-CRP) is a key biomarker for cardiovascular disease (CVD) risk.
• Current detection methods face challenges in point-of-care applications regarding sensitivity, speed, and convenience.
• A need exists for rapid, accurate, and user-friendly hs-CRP detection at the point of care.

Purpose of the Study:

• To develop a novel paper-based enzyme-linked immunosorbent assay (ELISA) for hs-CRP detection.
• To achieve high sensitivity and quantitative measurement suitable for point-of-care testing (POCT).
• To validate the assay's performance against conventional methods using clinical samples.

Main Methods:

• Development of a paper-based ELISA utilizing chemiluminescence for signal detection.
• Quantitative analysis correlating chemiluminescence intensity with hs-CRP concentration.
• Validation using clinical samples and comparison with a commercial high-sensitivity ELISA kit.

Main Results:

• The assay demonstrated a linear correlation (r=0.999) between hs-CRP concentration and chemiluminescence.
• Achieved a low limit of detection (LOD) of 0.49 ng/mL, comparable to conventional ELISA and superior to many POCT methods.
• Showed excellent precision (intra-assay CV <7%, inter-assay CV <10%) and strong agreement with commercial kits (r=0.975).

Conclusions:

• The developed paper-based ELISA provides a sensitive, rapid, and convenient method for hs-CRP detection.
• This assay is suitable for point-of-care applications, requiring minimal sample volume (4 μL) and short assay time (<30 min).
• It holds potential as a pre-screening tool for identifying individuals at elevated risk of cardiovascular disease.