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Updated: Apr 23, 2026
Author Spotlight: Advancements and Challenges in Hepatitis B Virus Detection
Published on: December 15, 2023
Detection of Hepatitis B Virus Large Surface Protein Using a Time-Resolved Immunofluorometric Assay
Zhigang Hu1,2, Mei Li2, Jie Liu2
1Department of Laboratory Medicine, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China.
Insights
A new time-resolved immunofluorometric assay (TR-IFMA) offers superior sensitivity and a broader detection range for hepatitis B virus L protein compared to ELISA. This method aids in HBV screening and antiviral therapy monitoring.
Area of Science:
- Biochemistry
- Immunology
- Hepatology
Background:
- Hepatitis B virus (HBV) large surface protein (L protein) detection is crucial for diagnosis and treatment monitoring.
- Existing methods like ELISA have limitations in sensitivity and detection range.
Purpose of the Study:
- To develop and validate a novel time-resolved immunofluorometric assay (TR-IFMA) for enhanced detection of HBV L protein.
- To compare the performance of the developed TR-IFMA with the classical enzyme-linked immunosorbent assay (ELISA).
Main Methods:
- Development of a TR-IFMA assay for HBV L protein.
- Evaluation of assay precision, sensitivity, specificity, recovery, and stability.
- Comparative analysis against a commercial ELISA kit.
Main Results:
- TR-IFMA demonstrated significantly higher sensitivity (0.1 ng/ml) and specificity (up to 100%) than ELISA.
- The TR-IFMA exhibited a wider linear detection range (0.63-10,367 ng/ml) compared to ELISA (10.12-1095.9 ng/ml).
- High precision (intra- and interassay <5%), excellent recovery (101.17%), and improved stability were observed for TR-IFMA.
Conclusions:
- The developed TR-IFMA is a highly sensitive and robust assay for detecting HBV L protein.
- TR-IFMA offers a broader detection range and improved performance over conventional ELISA.
- This TR-IFMA holds significant potential for HBV screening and monitoring antiviral therapy effectiveness.
Background:
To establish a novel method based on time-resolved immunofluorometric assay (TR-IFMA) with higher sensitivity and a broader detection range for detecting serum hepatitis B virus large surface protein (L protein).
Methods:
The precision, sensitivity, specificity, coefficient of recovery, and stability of the assay were evaluated and comparison with the classical enzyme-linked immunosorbent assay (ELISA) was also executed.
Results:
The precision, specificity, and sensitivity of the TR-IFMA were clearly better than ELISA. Particularly, the sensitivity was 0.1 ng/ml; moreover, the specificity was 100%, 96%, 92.5%, 96.9%, 97.8%, and 100% in the sera of healthy blood donors, systemic lupus erythematosus (SLE) patients, rheumatoid arthritis (RA) patients, hepatitis C virus (HCV) patients, cytomegalovirus (CMV) infection patients, and pregnant patients, respectively. Meanwhile, we observed that the established TR-IFMA kit has a wider acceptable linear range of 0.63-10,367 ng/ml rather than the regular commercial ELISA kit having range of only 10.12-1095.9 ng/ml. Subsequently, correlation coefficient between the TR-IFMA and ELISA was 0.8009. The intra- and interassay precision rates were less than 5% for three different concentrations. The average recovery rate for L protein was 101.17%. In sum, the established assay kit performed better in terms of stability than the commercial ELISA kit.
Conclusion:
The TR-IFMA that we developed for L protein presented a higher sensitivity and wider detecting range than regular commercial ELISA. Therefore, this TR-IFMA has promising value both in the screening of HBV and monitoring of antiviral therapy.
