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Immunochemiluminometric assay for hepatitis B surface antigen

H R Schroeder, C M Hines, D D Osborn

    Clinical Chemistry
    |August 1, 1981
    PubMed
    Summary
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    A new chemiluminescence immunoassay for hepatitis B surface antigen offers a sensitive and specific alternative to radioassays. This non-radioisotopic method uses stable reagents and automated readout for reliable hepatitis B diagnosis.

    Area of Science:

    • Clinical Chemistry
    • Immunology
    • Hepatitis Diagnostics

    Background:

    • Hepatitis B virus (HBV) infection is a significant global health concern.
    • Accurate detection of hepatitis B surface antigen (HBsAg) is crucial for diagnosis and management.
    • Existing diagnostic methods, such as radioimmunoassay (RIA), present challenges with reagent stability and handling.

    Purpose of the Study:

    • To describe a novel "sandwich" immunoassay for detecting hepatitis B surface antigen (HBsAg).
    • To evaluate the performance of this assay using chemiluminescence detection.
    • To offer a stable, non-radioisotopic alternative for HBsAg quantification.

    Main Methods:

    • Development of a "sandwich" immunoassay utilizing an antibody-coated microtitration plate.

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  • Incubation of test specimens (100 µL) with antibodies to capture HBsAg.
  • Detection of bound antigen using a second antibody labeled with an isoluminol derivative, followed by chemiluminescence measurement.
  • Automated readout of chemiluminescence using an in-house luminometer with microperoxidase and peroxide.

    • Main Results:

    • The chemiluminescence immunoassay demonstrated comparable specificity and sensitivity to a radioimmunoassay procedure.
    • Assay variability was low, with within- and between-assay coefficients of variation (CV) ranging from 7% to 13%.
    • Reagents exhibited excellent stability, remaining effective at 4°C for several months.

    Conclusions:

    • The developed chemiluminescence immunoassay is a viable, sensitive, and specific method for HBsAg detection.
    • This non-radioisotopic assay overcomes the stability and convenience issues associated with radioactive labels.
    • The assay provides a reliable and stable diagnostic tool for hepatitis B infection.