酶免疫测定法与时间分辨荧光免疫测定法的敏感性和特异性比较。

A comparison of the sensitivity and specificity of enzyme immunoassays and time-resolved fluoroimmunoassay.

作者信息

Roberts I M, Jones S L, Premier R R, Cox J C

机构信息

Commonwealth Serum Laboratories Limited, Parkville, Victoria, Australia.

出版信息

J Immunol Methods. 1991 Sep 20;143(1):49-56. doi: 10.1016/0022-1759(91)90271-g.

DOI:10.1016/0022-1759(91)90271-g

PMID:1919036

Abstract

Time-resolved fluoroimmunoassay (TR-FIA) and various enzyme immunoassays (EIA) were compared in order to determine the detection system which showed the greatest degree of sensitivity without sacrificing specificity. The system chosen for the evaluation of these assays was the detection of antibodies to human immunodeficiency virus (HIV). For EIA, horseradish peroxidase (HRP) and alkaline phosphatase (AP) were investigated, each with a number of different substrates. HRP with its fluorogenic substrate, 3-(p-hydroxyphenyl)propionic acid (HPPA) was 1.6 times (p less than 0.01) more sensitive than with 3,3',5,5'-tetramethylbenzidine (TMB) and four times (p less than 0.001) more sensitive than with 2,2'-azino-di(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS). AP with its fluorogenic substrate, 4-methylumbelliferyl phosphate (4MeUP), was 6-7 times (p less than 0.001) more sensitive than with phenolphthalein monophosphate (PMP) and 8-13 times (p less than 0.001) more sensitive than with p-nitrophenyl phosphate (pNPP). TR-FIA with Eu3(+)-labelled anti-human IgG was equivalent in sensitivity to HRP with TMB and AP with 4MeUP.

摘要

为了确定在不牺牲特异性的情况下显示出最高灵敏度的检测系统，对时间分辨荧光免疫分析（TR-FIA）和各种酶免疫分析（EIA）进行了比较。选择用于评估这些分析的系统是检测针对人类免疫缺陷病毒（HIV）的抗体。对于EIA，研究了辣根过氧化物酶（HRP）和碱性磷酸酶（AP），每种都有许多不同的底物。HRP与其荧光底物3-(对羟基苯基)丙酸（HPPA）相比，灵敏度比使用3,3',5,5'-四甲基联苯胺（TMB）时高1.6倍（p<0.01），比使用2,2'-叠氮基-二(3-乙基苯并噻唑啉-6-磺酸)（ABTS）时高4倍（p<0.001）。AP与其荧光底物4-甲基伞形酮磷酸酯（4MeUP）相比，灵敏度比使用酚酞单磷酸酯（PMP）时高6-7倍（p<0.001），比使用对硝基苯基磷酸酯（pNPP）时高8-13倍（p<0.001）。用Eu3(+)标记的抗人IgG的TR-FIA在灵敏度上与使用TMB的HRP和使用4MeUP的AP相当。