Nielsen K, Smith P, Yu W L, Elmgren C, Nicoletti P, Perez B, Bermudez R, Renteria T
Canadian Food Inspection Agency, Ottawa Laboratory Fallowfield, 3851 Fallowfield Road, Ottawa, Ontario, Canada K2H 8P9.
Vet Microbiol. 2007 Sep 20;124(1-2):173-7. doi: 10.1016/j.vetmic.2007.03.023. Epub 2007 Mar 30.
A second generation competitive enzyme immunoassay (CELISA) for detection of bovine antibody to Brucella abortus was developed. This assay was different from previously developed CELISAs in that the detection reagent used was a recombinant combination of the receptor portions of protein A and protein G, labelled with horseradish peroxidase. This eliminates the need for polyclonal anti-mouse-enzyme conjugate reagents for detection thus allowing for true standardization. The assay utilized a monoclonal antibody specific for a common epitope of the O-polysaccharide (OPS) of smooth lipopolysaccharide (SLPS) derived from B. abortus S1119.3 but which did not react with protein A/G. This monoclonal antibody was used to compete with antibody in the bovine test serum. Binding of bovine antibody to the smooth lipopolysaccharide antigen was then measured directly with the protein A/G enzyme conjugate. In this case, development of colour in the reaction was indicative of the presence of bovine antibody. The performance characteristics, sensitivity, specificity and exclusion of B. abortus S19 vaccinated animals, of the assay were very similar to those of the classical CELISA.
开发了一种用于检测牛抗布鲁氏菌流产抗体的第二代竞争性酶免疫测定法（CELISA）。该测定法与先前开发的CELISA不同之处在于，所使用的检测试剂是用辣根过氧化物酶标记的蛋白A和蛋白G受体部分的重组组合。这消除了检测中使用多克隆抗小鼠酶缀合试剂的需求，从而实现了真正的标准化。该测定法使用了一种单克隆抗体，该抗体对源自布鲁氏菌流产S1119.3的光滑脂多糖（SLPS）的O-多糖（OPS）的共同表位具有特异性，但不与蛋白A/G反应。该单克隆抗体用于与牛测试血清中的抗体竞争。然后用蛋白A/G酶缀合物直接测量牛抗体与光滑脂多糖抗原的结合。在这种情况下，反应中颜色的显现表明存在牛抗体。该测定法的性能特征、敏感性、特异性以及对接种布鲁氏菌流产S19疫苗动物的排除情况与经典CELISA非常相似。
