Lim P L, Ko K H
Department of Microbiology, University of Hong Kong.
J Immunol Methods. 1990 Dec 31;135(1-2):9-14. doi: 10.1016/0022-1759(90)90250-y.
A simple two stage assay was developed for the detection of IgM antibodies to either one of two microorganisms chosen arbitrarily for this study. Salmonella enteritidis and Trichinella spiralis. In the first stage, magnetic polystyrene beads (Dynabeads) coated with anti-mu (mouse) antibodies were incubated with the test material for 45 min to capture the IgM antibodies. In the second stage, indicator latex particles were incubated with the Dynabeads for 30 min and the results read following settlement of the Dynabeads under the influence of a magnet. Two types of indicator particles were used: blue-coloured (sensitized with Trichinella antigen) and red-coloured (sensitized with Salmonella antigen). These were mixed in suitable proportions to form a purple suspension. Reaction of either type of latex particles due to binding to the adsorbed IgM antibodies resulted in the settlement of that particle and hence a change of colour in the suspension to either red (if Trichinella-specific antibodies alone were present) or blue (if Salmonella-specific antibodies alone were present). When applied to the sera (used at 1/10 dilution) of both normal and immunized mice, the assay was positive for all but one (18) immune sera, and negative for all but one (9) normal sera.
我们开发了一种简单的两阶段检测方法,用于检测针对本研究中任意选择的两种微生物之一的 IgM 抗体。这两种微生物分别是肠炎沙门氏菌和旋毛虫。在第一阶段,将包被有抗 μ(小鼠)抗体的磁性聚苯乙烯珠(Dynabeads)与测试材料孵育 45 分钟,以捕获 IgM 抗体。在第二阶段,将指示乳胶颗粒与 Dynabeads 孵育 30 分钟,然后在磁铁的作用下 Dynabeads 沉降后读取结果。使用了两种类型的指示颗粒:蓝色(用旋毛虫抗原致敏)和红色(用沙门氏菌抗原致敏)。将它们按适当比例混合形成紫色悬浮液。由于与吸附的 IgM 抗体结合,任何一种乳胶颗粒发生反应都会导致该颗粒沉降,从而使悬浮液颜色变为红色(如果仅存在旋毛虫特异性抗体)或蓝色(如果仅存在沙门氏菌特异性抗体)。当将该检测方法应用于正常小鼠和免疫小鼠的血清(以 1/10 稀释使用)时,除一份(18 份)免疫血清外,所有检测均呈阳性,除一份(9 份)正常血清外,所有检测均呈阴性。