Libby J M, Wada H G
Molecular Devices Corporation, Menlo Park, California 94025.
J Clin Microbiol. 1989 Jul;27(7):1456-9. doi: 10.1128/jcm.27.7.1456-1459.1989.
A light-addressable potentiometric (silicon) sensor was used in an immunofiltration procedure for the detection of pathogenic bacteria. Yersinia pestis was detected by filtering the cells onto nitrocellulose membranes and then filtering anti-Y. pestis mouse monoclonal antibody and anti-mouse immunoglobulin G-horseradish peroxidase conjugate. For Neisseria meningitidis detection, mouse monoclonal antibody to the major outer membrane protein of this bacterium was coupled directly to horseradish peroxidase. N. meningitidis cell suspensions were filtered onto polycarbonate membranes, and the enzyme conjugate was allowed to react with the filtered bacteria. The presence of both enzyme conjugates was determined potentiometrically with the silicon sensor. The sensitivity of this technique relative to that of an enzyme-linked immunosorbent assay for N. meningitidis was determined. Fewer than 1,000 bacterial cells could be detected with the silicon sensor in a 20-min assay, whereas a 2.5-h enzyme-linked immunosorbent assay with the same antigen and antibody preparations was significantly less sensitive.
一种光寻址电位(硅)传感器被用于免疫过滤程序中以检测病原菌。通过将细胞过滤到硝酸纤维素膜上,然后过滤抗鼠疫耶尔森菌小鼠单克隆抗体和抗小鼠免疫球蛋白G-辣根过氧化物酶缀合物来检测鼠疫耶尔森菌。对于脑膜炎奈瑟菌的检测,针对该细菌主要外膜蛋白的小鼠单克隆抗体直接与辣根过氧化物酶偶联。将脑膜炎奈瑟菌细胞悬液过滤到聚碳酸酯膜上,并使酶缀合物与过滤后的细菌反应。用硅传感器以电位法测定两种酶缀合物的存在情况。测定了该技术相对于脑膜炎奈瑟菌酶联免疫吸附测定法的灵敏度。在20分钟的检测中,硅传感器能够检测到少于1000个细菌细胞,而使用相同抗原和抗体制剂进行的2.5小时酶联免疫吸附测定法的灵敏度则明显较低。