Umeda M, Tomita T, Shibata H, Seki M, Yasuda T
Laboratory of Biological Products, University of Tokyo, Japan.
J Clin Microbiol. 1988 May;26(5):804-7. doi: 10.1128/jcm.26.5.804-807.1988.
We developed a liposome lysis assay for determining anti-streptolysin O antibodies (ASO) in human sera involving the use of carboxyfluorescein-entrapped multilamellar liposomes. This assay system was based on the inhibition of streptolysin O-induced liposome lysis by ASO. Briefly, after incubation of a given amount of streptolysin O with ASO for 30 min at 37 degrees C, carboxyfluorescein-entrapped liposomes composed of egg yolk phosphatidylcholine and cholesterol in a molar ratio of 1:1 were added to the mixture to determine the residual streptolysin O activity. Liposome lysis, detected as carboxyfluorescein release from the liposomes, was inversely proportional to the ASO titer. The results of within-run and between-run precision studies indicated that the liposome lysis assay is accurate and gives reproducible data. Bilirubin, hemoglobin, and triglycerides did not interfere with the liposome lysis assay. The ASO titers of 100 patient sera, evaluated by our new method and the Rantz-Randall method, showed good correlation.
我们开发了一种脂质体裂解测定法，用于测定人血清中的抗链球菌溶血素O抗体（ASO），该方法涉及使用包裹羧基荧光素的多层脂质体。该测定系统基于ASO对链球菌溶血素O诱导的脂质体裂解的抑制作用。简而言之，将一定量的链球菌溶血素O与ASO在37℃下孵育30分钟后，向混合物中加入由蛋黄磷脂酰胆碱和胆固醇按摩尔比1:1组成的包裹羧基荧光素的脂质体，以测定残留的链球菌溶血素O活性。脂质体裂解以脂质体中羧基荧光素的释放来检测，与ASO滴度成反比。批内和批间精密度研究结果表明，脂质体裂解测定法准确且数据可重复。胆红素、血红蛋白和甘油三酯不干扰脂质体裂解测定法。用我们的新方法和兰茨-兰德尔方法评估的100份患者血清的ASO滴度显示出良好的相关性。
