Smith K O, Ludwig M J, Stigall B W, Boswell N
Department of Microbiology, University of Texas Health Science Center, San Antonio 78284-7758.
J Immunol Methods. 1991 Feb 15;136(2):239-46. doi: 10.1016/0022-1759(91)90010-d.
An enzyme-linked immunosorbent assay (ELISA) technique is described which utilizes a commercially available glass microscope slide coated with hydrophobic teflon in such a pattern as to give 30 small circular wells, each of which has a glass bottom. Each well serves as a solid phase, analogous to a microtiter well for adsorption of purified human immunodeficiency virus (HIV) antigens. Since only 5-10 microliter volumes of reagents are used and rinsing processing is simple, the cost per test is much less than most other ELISA technologies. HIV antigen is stable for over 1 year at 37 degrees C when dried on the glass slides. The sensitivity and specificity of the micro slide immunoenzymatic assay (Micro-SIA) was studied by testing randomly selected, known HIV-seropositive and seronegative plasma. Results compare well with microtiter and Western blot assays. A simple vertical-beam colorimeter is described (useful in the Micro-SIA) which can be easily assembled by the user from commonly available components.
本文描述了一种酶联免疫吸附测定(ELISA)技术,该技术使用一种涂有疏水性聚四氟乙烯的市售玻璃显微镜载玻片,其图案可形成30个小圆形孔,每个孔都有玻璃底部。每个孔作为固相,类似于微量滴定孔,用于吸附纯化的人类免疫缺陷病毒(HIV)抗原。由于每次检测仅使用5-10微升试剂,且冲洗过程简单,因此每次检测的成本远低于大多数其他ELISA技术。HIV抗原在37摄氏度下干燥在玻璃载玻片上时可稳定保存1年以上。通过检测随机选择的已知HIV血清阳性和血清阴性血浆,研究了微量载玻片免疫酶测定(Micro-SIA)的灵敏度和特异性。结果与微量滴定法和蛋白质印迹法相比效果良好。本文还描述了一种简单的垂直光束比色计(可用于Micro-SIA),用户可以很容易地用常见组件组装而成。