Enhanced chemiluminescence ELISA for the detection of antibody to hepatitis B virus surface antigen.
A chemiluminescent enzyme linked immunosorbent assay (ELISA) for the detection of antibody to hepatitis B virus surface antigen (anti-HBs) in human serum has been developed. Polystyrene microtitre plates were coated with recombinant, yeast-derived hepatitis B surface antigen (rec-HBsAg). Patient serum samples and appropriate controls were added to the rec-HBsAg-coated wells and incubated to bind anti-HBs. The wells were then washed and a fluorescein isothiocyanate (FITC) conjugate of a human plasma-derived hepatitis B surface antigen (HBsAg) was added. Following incubation and further washing, the bound FITC-labelled HBsAg was detected after addition of a horseradish peroxidase (HRP) conjugate of a monoclonal anti-FITC antibody and assaying for the enzyme. The activity of the HRP was measured using luminol and hydrogen peroxide as substrates and iodophenol as a chemiluminescence enhancer. The luminescence was recorded using a camera luminometer. Preliminary tests have shown the assay to be suitable for the detection of antibody in sera from both vaccinees and also from individuals with a past hepatitis B virus infection. The use of the FITC-anti-FITC system together with the measurement of a chemiluminescence signal makes possible the completion of this assay in a few hours. The assay has been shown to be both specific and sensitive and provides a permanent photographic record.
Ireland
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Division of Microbiological Reagents and Quality Control, Central Public Health Laboratory, Colindale, London, UK.
Samuel
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D003124:Colorimetry; D004797:Enzyme-Linked Immunosorbent Assay; D006509:Hepatitis B / Q000175:diagnosis; D006510:Hepatitis B Antibodies / Q000032:analysis*; D006514:Hepatitis B Surface Antigens; D006801:Humans; D008163:Luminescent Measurements; D011863:Radioimmunoassay; D014761:Viral Hepatitis Vaccines / Q000627:therapeutic use
D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D014761:Viral Hepatitis Vaccines