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一种用于检测特异性IgM抗体的双颗粒比浊乳胶免疫测定法。

A two-particle turbidometric latex immunoassay for the detection of specific IgM antibodies.

作者信息

Lim P L, Ko K H, Choy W F

机构信息

Department of Microbiology, University of Hong Kong.

出版信息

J Immunol Methods. 1989 Feb 24;117(2):267-73. doi: 10.1016/0022-1759(89)90149-x.

Abstract

A simple two stage assay using latex particles as a reaction indicator has been developed for the detection of IgM antibodies to Trichinella spiralis. In the first stage, magnetic polystyrene beads (Dynabeads) coated with T. spiralis antigen were incubated for 30 min with the test serum. After washing, in the second stage, the assay was developed for 1 h using anti-mu-coated latex particles. After sedimentation of the Dynabeads the turbidity of the resultant latex suspension was measured spectrophotometrically at a wavelength of 400 nm. A decrease in turbidity of more than 20% from that of the control, unreacted, suspension was considered positive. Using an IgM phosphorylcholine-binding monoclonal antibody which was reactive with T. spiralis, the sensitivity of the assay was determined to be 110 ng/ml of antibody. This was 20-fold less than the sensitivity achieved in an indirect enzyme-linked immunosorbent assay (ELISA). When the assay was applied to sera obtained from CBA/N or BALB/c mice, which were either normal or immunized against T. spiralis, the expected results were obtained with titers up to 1/640 observed, and confirmed (r = 0.93, P less than 0.001) in the ELISA.

摘要

已开发出一种简单的两阶段检测方法,使用乳胶颗粒作为反应指示剂来检测抗旋毛虫的IgM抗体。在第一阶段,将包被有旋毛虫抗原的磁性聚苯乙烯珠(Dynabeads)与待测血清孵育30分钟。洗涤后,在第二阶段,使用抗μ包被的乳胶颗粒进行1小时的检测。Dynabeads沉淀后,在400nm波长下用分光光度法测量所得乳胶悬浮液的浊度。与未反应的对照悬浮液相比,浊度降低超过20%被视为阳性。使用与旋毛虫反应的IgM磷酸胆碱结合单克隆抗体,该检测方法的灵敏度测定为110ng/ml抗体。这比间接酶联免疫吸附测定(ELISA)所达到的灵敏度低20倍。当该检测方法应用于从CBA/N或BALB/c小鼠获得的血清时,这些小鼠要么正常,要么已针对旋毛虫进行免疫,得到了预期的结果,观察到的滴度高达1/640,并在ELISA中得到证实(r = 0.93,P小于0.001)。

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