Achacha M, Mittal K R
Département de Pathologie et Microbiologie, Université de Montréal, Québec, Canada.
Vet Microbiol. 1996 Jan;48(1-2):73-85. doi: 10.1016/0378-1135(95)00129-8.
Four murine monoclonal antibodies (mAbs) designated as C9E8, A10, G12, and G8 which recognized both Serpulina hyodysenteriae and S. innocens were produced and characterized. The mAbs reacted with whole cell antigens in ELISA, indirect immunofluorescence and immunoblot assays. The mAbs did not show any cross reactivity in rapid dot ELISA or immunoblot assay with Leptospira icterohemorrhagiae, Campylobacter jejuni and Escherichia coli. Treatment of whole cell suspension with proteinase K and sodium periodate indicated that the reacting epitopes of the mAbs were protein in nature. The genus-specific antigens were identified as heat-stable proteins with molecular weight in the range of 26 to 45 kDa. Immunofluorescence and immunogold labelling studies showed that the antibody-binding epitopes were exposed on the outer-surface of the spirochaetal cell wall. The mAbs inhibited growth of reference strains of both S. hyodysenteriae and S. innocens in vitro but failed to cause agglutination. The detection of spirochaetal forms directly in fecal smears or paraffin-embeded tissue sections from experimentally infected pigs indicated that such mAbs were potentially useful for the diagnosis of swine spirochaetosis. This is the first report of mAbs identifying and characterizing common antigens of porcine Serpulina.
制备并鉴定了四种鼠单克隆抗体(mAb),分别命名为C9E8、A10、G12和G8,它们能识别猪痢疾蛇形螺旋体和无害蛇形螺旋体。这些单克隆抗体在酶联免疫吸附测定(ELISA)、间接免疫荧光和免疫印迹分析中与全细胞抗原发生反应。在快速斑点ELISA或免疫印迹分析中,这些单克隆抗体与出血性钩端螺旋体、空肠弯曲菌和大肠杆菌均未显示任何交叉反应。用蛋白酶K和高碘酸钠处理全细胞悬液表明,单克隆抗体的反应表位本质上是蛋白质。属特异性抗原被鉴定为分子量在26至45 kDa范围内的热稳定蛋白。免疫荧光和免疫金标记研究表明,抗体结合表位暴露在螺旋体细胞壁的外表面。这些单克隆抗体在体外抑制猪痢疾蛇形螺旋体和无害蛇形螺旋体参考菌株的生长,但未能引起凝集反应。直接在实验感染猪的粪便涂片或石蜡包埋组织切片中检测螺旋体形态表明,此类单克隆抗体对猪螺旋体病的诊断具有潜在用途。这是关于单克隆抗体鉴定和表征猪蛇形螺旋体共同抗原的首次报道。