一种利用沟虾生物发光的人绒毛膜促性腺激素固相发光免疫测定法。

A solid-phase luminescent immunoassay for human chorionic gonadotropin using Pholas dactylus bioluminescence.

作者信息

Longeon A, Henry J P, Henry R

出版信息

J Immunol Methods. 1986 May 1;89(1):103-9. doi: 10.1016/0022-1759(86)90037-2.

Abstract

A solid-phase luminescent immunoassay (LIA), based on the light emission produced as a result of the oxidation of Pholas dactylus luciferin by horseradish peroxidase (HRP) in the presence of molecular oxygen, was developed for human chorionic gonadotropin (hCG). The light emitted in the presence of diethyldithiocarbamate permitted the detection of 0.1 fmol HRP. HRP retained most of its light-emitting capacity after coupling with purified anti-hCG antibody by glutaraldehyde. The LIA involved immobilization of the antigen in plastic tubes coated with purified anti-hCG antibody and detection of the immunocomplex by light emission in the presence of Pholas luciferin. Light emission was linear for antigen concentration within the range 0.5-100 ng/ml. LIA correlates reasonably well with RIA and has a comparable sensitivity (0.5 ng hCG/ml).

摘要

基于辣根过氧化物酶（HRP）在分子氧存在下氧化紫贻贝荧光素产生的发光现象，开发了一种用于检测人绒毛膜促性腺激素（hCG）的固相发光免疫分析（LIA）方法。在二乙基二硫代氨基甲酸盐存在下发出的光可检测到0.1 fmol的HRP。通过戊二醛将HRP与纯化的抗hCG抗体偶联后，HRP保留了大部分发光能力。该LIA方法包括将抗原固定在包被有纯化抗hCG抗体的塑料管中，并在紫贻贝荧光素存在下通过发光检测免疫复合物。抗原浓度在0.5-100 ng/ml范围内时，发光呈线性。LIA与放射免疫分析（RIA）的相关性较好，灵敏度相当（0.5 ng hCG/ml）。