Enzyme-linked immunosorbent assay for hepatitis B surface antigen

Area of Science:

• Hepatology
• Immunology
• Diagnostic Assays

Background:

• Hepatitis B virus (HBV) infection remains a significant global health concern.
• Accurate and rapid detection of hepatitis B surface antigen (HBSAg) is crucial for diagnosis and management.
• Existing diagnostic methods have limitations in sensitivity, speed, or accessibility.

Purpose of the Study:

• To develop and evaluate a solid-phase enzyme-linked immunosorbent assay (ELISA) for detecting HBSAg.
• To assess the sensitivity, specificity, and practicality of the developed ELISA method.
• To compare the performance of ELISA with established diagnostic techniques.

Main Methods:

• Development of a "sandwich" principle solid-phase ELISA using microtiter plates.
• Evaluation of detection levels, with a sensitivity threshold of ≤5-10 ng/ml HBSAg.
• Comparative analysis with radioimmunoassay (RIA) and reversed hemagglutination (RHA) tests.
• Clinical validation across 11 European medical centers involving over 50,000 samples.

Main Results:

• The ELISA method allows for results to be obtained within one day, readable by eye or colorimeter.
• ELISA demonstrated comparable sensitivity to RIA in dilution series and patient studies.
• In a large-scale multicenter study, ELISA detected significantly more HBSAg-positive samples than the RHA test.
• An average of 2.2% false-positive reactions were observed, managed with a confirmatory test.

Conclusions:

• The developed ELISA is a sensitive and practical method for HBSAg detection.
• ELISA offers advantages in speed and detection rates compared to RHA, with performance comparable to RIA.
• This assay provides a valuable tool for hepatitis B diagnosis in clinical settings.