Sciortino C V
Department of Microbiology, Veterans Administration Medical Center, Louisville, KY 40206.
Hybridoma. 1993 Jun;12(3):333-42. doi: 10.1089/hyb.1993.12.333.
We prepared a murine monoclonal antibody that recognized the Helicobacter pylori urease. Monoclonal antibody, U2, reacted with purified native urease in enzyme linked immunosorbent assay but it did not react in Western immunoblot analysis. Urease was immunoprecipitated with U2 bound to Staphylococcus aureus protein A, followed by elution and visualization by sodium dodecyl-sulfate polyacrylamide gel electrophoresis. The species and strain specificity of U2 were determined by biodot reaction with H. pylori, H. mustelae, Campylobacter spp., and other gram-negative bacteria. Monoclonal antibody, U2, was used for an indirect immunofluorescent stain of formalin fixed gastric biopsies from patients with H. pylori gastritis. The immunofluorescent stain showed spiral and coccoid forms of H. pylori within the gastric mucosa. U2 was specific for H. pylori and did not react with other tested bacteria. These findings suggest that antibody U2 might be of diagnostic value for specific immunofluorescence detection of H. pylori.
我们制备了一种识别幽门螺杆菌脲酶的鼠单克隆抗体。单克隆抗体U2在酶联免疫吸附测定中与纯化的天然脲酶发生反应,但在蛋白质免疫印迹分析中不发生反应。脲酶与结合在金黄色葡萄球菌蛋白A上的U2进行免疫沉淀,然后洗脱并通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳进行可视化分析。通过与幽门螺杆菌、鼬螺杆菌、弯曲菌属及其他革兰氏阴性菌的斑点杂交反应来确定U2的种属和菌株特异性。单克隆抗体U2用于对幽门螺杆菌胃炎患者的福尔马林固定胃活检组织进行间接免疫荧光染色。免疫荧光染色显示胃黏膜内幽门螺杆菌的螺旋形和球形形态。U2对幽门螺杆菌具有特异性,与其他测试细菌不发生反应。这些发现表明抗体U2可能对幽门螺杆菌的特异性免疫荧光检测具有诊断价值。