Rapid detection of hepatitis B surface antigen by double antibody radioimmunoassay

✍ Scribed by C. J. Burrell; G. Leadbetter; S. H. Black; W. M. Hunter

• Publisher: John Wiley and Sons
• Year: 1978
• Tongue: English
• Weight: 416 KB
• Volume: 3
• Category: Article
• ISSN: 0146-6615
• DOI: 10.1002/jmv.1890030106

✦ Synopsis

Abstract

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.