An improved method for rapid identification of hook effect samples in HBsAg quantitative assay

Area of Science:

• Clinical Chemistry
• Immunodiagnostics
• Hepatitis B Virus Research

Background:

• Quantitative hepatitis B surface antigen (HBsAg) assays are crucial for diagnosing hepatitis B virus (HBV) infection.
• A significant challenge in HBsAg quantification is the 'hook effect,' where high antigen levels paradoxically yield false-negative results, compromising diagnosis.

Purpose of the Study:

• To develop and validate a rapid, improved method for detecting and resolving the hook effect in quantitative HBsAg assays.
• To enhance the reliability and accuracy of HBsAg testing on a chemiluminescence platform.

Main Methods:

• Three methods were evaluated on the LiCA® 500 instrument using hook effect and non-hook effect samples.
• Method 1: Current two-step procedure (25 min total reaction time).
• Method 3: Improved one-step procedure with reduced reagent volume (130 µL) and significantly shortened reaction time (1 min).

Main Results:

• Method 3 successfully differentiated hook effect samples in just 1 minute, achieving results comparable to the 25-minute Method 2.
• The improved method leverages competitive inhibition for clearer distinction of hook effect samples.
• Method 3 demonstrated superior performance in resolving hook effect issues compared to the standard assay.

Conclusions:

• A rapid, one-step method (Method 3) effectively identifies hook effect samples in HBsAg quantification.
• This optimized procedure offers a significant improvement for chemiluminescence-based HBsAg assays, enhancing diagnostic capabilities.
• The findings suggest Method 3 is a valuable advancement for accurate hepatitis B diagnosis.