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Highly sensitive hepatitis B surface antigen detection by measuring stable nitroxide radical formation with ESR
T Matsuo1, H Shinzawa, H Togashi
1The Second Department of Internal Medicine, Yamagata University School of Medicine, Japan.
Free Radical Biology & Medicine
|December 5, 1998
Summary
A new, highly sensitive electron spin resonance (ESR) method using p-acetamidophenol (p-AP) and 4-hydrazonomethyl-1-hydroxy-2,2,5,5,-tetramethyl-3-imidazoline-3-o xide (HHTIO) can detect hepatitis B surface antigen (HBsAg) with greater accuracy than ELISA. This advancement is crucial for identifying hepatitis B virus (HBV) carriers and improving blood screening.
Area of Science:
- Biochemistry
- Medical Diagnostics
- Virology
Background:
- Hepatitis B virus (HBV) infection remains a significant global health concern, particularly in endemic regions.
- Current screening methods like ELISA for hepatitis B surface antigen (HBsAg) have limitations, leading to potential transmission via blood transfusion and undetected carriers.
- There is a critical need for more sensitive diagnostic tools to accurately detect HBsAg and prevent HBV spread.
Purpose of the Study:
- To develop and validate a highly sensitive method for detecting HBsAg.
- To compare the sensitivity and reproducibility of the new method against the standard ELISA.
- To assess the utility of the new method in identifying potential HBV carriers missed by conventional screening.
Main Methods:
- Development of a novel HBsAg detection assay utilizing electron spin resonance (ESR) spectroscopy.
- The method relies on measuring peroxidase activity via the formation of stable nitroxide radicals using p-acetamidophenol (p-AP) and 4-hydrazonomethyl-1-hydroxy-2,2,5,5,-tetramethyl-3-imidazoline-3-o xide (HHTIO).
- Establishment of cut-off values using sera from healthy adults and HBsAg-positive individuals, followed by validation on samples with borderline ELISA results and non-B, non-C hepatitis cases.
Main Results:
- The p-AP/HHTIO-ESR method demonstrated significantly higher sensitivity (approximately 10 times) compared to standard ELISA for HBsAg detection.
- Excellent reproducibility was observed with the developed ESR-based assay.
- The method successfully identified HBsAg in individuals with negative ELISA results but high cut-off index values, and in some non-B, non-C hepatitis patients, with some of these cases also testing positive for HBV DNA via PCR.
Conclusions:
- The developed p-AP/HHTIO-ESR method offers a highly sensitive and practical approach for HBsAg detection.
- This assay shows promise for improved screening of HBV carriers and blood donations, potentially surpassing the capabilities of current ELISA techniques.
- The sensitivity of this ESR-based method is comparable to PCR, highlighting its potential to enhance diagnostic accuracy in HBV screening programs.