Chemiluminescence enzyme immunoassay for thyroxin with use of glucose oxidase and a bis(2,4,6-trichlorophenyl)oxalate-fluorescent dye system.

Abstract

In this chemiluminescence enzyme immunoassay for thyroxin, glucose oxidase (EC 1.1.3.4) is the enzyme label and the bound and free fractions are separated by the double-antibody solid-phase technique. In the assay of enzyme activity, hydrogen peroxide generated from glucose substrate is measured by its chemiluminescence reaction with bis(2,4,6-trichlorophenyl)oxalate and 8-anilino-1-naphthalene-sulfonic acid. The measurable range of thyroxin is 2.5 to 200 micrograms/L. The coefficients of variation for thyroxin concentrations of 43.2 and 12.8 micrograms/L were 7.0% and 8.6% (intra-assay), and 6.4% and 12.3% (interassay), respectively. The proposed method is applicable to large-scale preliminary screening of neonates for congenital hypothyroidism, with samples consisting of dried blood spotted on filter paper.