使用细菌荧光素酶对血清中总胆汁酸进行生物发光测定。

Bioluminescent assay for total bile acids in serum with use of bacterial luciferase.

作者信息

Styrélius I, Thore A, Björkhem I

出版信息

Clin Chem. 1983 Jun;29(6):1123-7.

Abstract

This simple, rapid, sensitive kinetic bioluminescent method for the assay of bile acids in serum involves use of 3 alpha-hydroxysteroid dehydrogenase (EC 1.1.1.50) in combination with a new, commercially available NADH Monitoring Reagent (LKB-Wallac) containing a low activity of NADH:FMN-oxidoreductase and a high activity of bacterial luciferase. Interfering dehydrogenases in serum are inactivated in the test tube with trichloroacetic acid before the assay. The standard curve is linear for concentrations of bile acids up to about 300 mumol/L. With a sample volume of 20 microL, the detection limit is about 0.2 mumol/L. The within-run precision (CV) is about 10%, both at high and low concentrations of bile acids in serum. Correlation is good (r = 0.996) between results by this method and an enzymatic method based on spectrophotometry. However, the latter method is considerably less sensitive, and it is less precise at low concentrations of bile acids.

摘要

这种用于测定血清中胆汁酸的简单、快速、灵敏的动力学生物发光方法，涉及使用3α-羟基类固醇脱氢酶（EC 1.1.1.50），并结合一种新的、市售的NADH监测试剂（LKB-沃拉），该试剂含有低活性的NADH：FMN氧化还原酶和高活性的细菌荧光素酶。在测定前，血清中的干扰脱氢酶在试管中用三氯乙酸灭活。胆汁酸浓度高达约300μmol/L时，标准曲线呈线性。进样量为20μL时，检测限约为0.2μmol/L。血清中高、低浓度胆汁酸时，批内精密度（CV）约为10%。该方法与基于分光光度法的酶法结果之间相关性良好（r = 0.996）。然而，后一种方法灵敏度明显较低，且在低浓度胆汁酸时精密度较低。