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Rapid detection of hepatitis B surface antigen by double antibody radioimmunoassay
Author(s) -
Burrell C. J.,
Leadbetter G.,
Black S. H.,
Hunter W. M.
Publication year - 1978
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.1890030106
Subject(s) - radioimmunoassay , hbsag , chromatography , immunoradiometric assay , standard curve , antibody , detection limit , chemistry , virology , hepatitis b virus , immunology , biology , biochemistry , virus
HBsAg and anti‐HBs can be detected by double‐antibody radioimmunoassay in a reliable routine overnight assay of sensitivity approaching that of solidphase immunoradiometric assay. The test requires careful standardisation and regular preparation of I 125 ‐HBsAg of appropriate quality, but otherwise uses only small quantities of commercially available reagents. In this paper, the reaction kinetics of the first incubation (test sample + limiting amount of rabbit anti‐HBs), second incubation (reaction mixture + I 125 ‐HBsAg) and precipitation reaction (reaction mixture + antirabbit globulin) were examined, to determine the minimum time required for the assay compatible with adequate sensitivity and user convenience; in addition, in order to reduce the time of radioactive counting, the maximum amount of I 125 ‐HBsAg per test compatible with a satisfactory assay was examined. It was concluded that, by terminating each reaction before equilibrium was reached, results could be obtained in 2–3 hours with a detection limit similar to the standard overnight procedure. To allow routine examination of large numbers of samples, the possibilities of complete automation of this liquid‐phase type of assay were discussed.