一种使用抗生物素蛋白-生物素分离系统和甲状腺素-过氧化物酶偶联物的总甲状腺素酶免疫测定法。

An enzymeimmunoassay for total thyroxine using avidin-biotin separation system and thyroxine-peroxidase conjugate.

作者信息

Akman S, Karaca L, Kutluay T

机构信息

Department of Biochemistry, School of Medicine, Gulhane Military Medical Academy, Ankara, Turkey.

出版信息

J Immunoassay. 1995 Aug;16(3):325-41. doi: 10.1080/15321819508013566.

DOI:10.1080/15321819508013566

PMID:7593653

Abstract

An avidin-biotin enzymeimmunoassay for total thyroxine in serum is described. Avidin was adsorbed to biotinylated bovine serum albumin coated tubes prepared with glutaraldehyde as coupling agent. In the enzymeimmunoassay, affinity purified biotinylated anti-thyroxine IgG, sample or standards, and thyroxine-horseradish peroxidase were simultaneously added to the avidin coated tubes. The bound enzymatic activity was then measured with o-phenylenediamine and H2O2. Results showed that the assay has good precision (within-assay CV% less than 10% and between assay 11.7% in hypo- and 6.9% in hyperthyroid range), good assay range (0-800 nmol/L), good sensitivity (4 nmol/L), and can be performed in 2.5 hours. The results obtained correlate well (r = 0.93) with those of an RIA.

摘要

本文描述了一种用于血清总甲状腺素的抗生物素蛋白-生物素酶免疫测定法。将抗生物素蛋白吸附到以戊二醛作为偶联剂制备的生物素化牛血清白蛋白包被管上。在酶免疫测定中，将亲和纯化的生物素化抗甲状腺素IgG、样品或标准品以及甲状腺素-辣根过氧化物酶同时加入到抗生物素蛋白包被管中。然后用邻苯二胺和过氧化氢测量结合的酶活性。结果表明，该测定法具有良好的精密度（批内变异系数在甲状腺功能减退范围内小于10%，在甲状腺功能亢进范围内批间为11.7%，批内为6.9%）、良好的测定范围（0-800 nmol/L）、良好的灵敏度（4 nmol/L），并且可以在2.5小时内完成。所得结果与放射免疫分析结果具有良好的相关性（r = 0.93）。