Rasberry U, Rosenbusch R F
Veterinary Medical Research Institute, College of Veterinary Medicine, Iowa State University, Ames 50011, USA.
Hybridoma. 1995 Oct;14(5):481-5. doi: 10.1089/hyb.1995.14.481.
Mycoplasma bovis is a causative agent of bovine mastitis, arthritis, and pneumonia. Six monoclonal antibodies (MAbs) against M. bovis were prepared and used to characterize specific antigens of the mycoplasma. Reactivity of the MAbs to six M. bovis strains was tested by IFA, ELISA, and immunoblotting. The specificity of these MAbs was tested by the same methods against 8 other species of bovine mycoplasmas and 1 mycoplasma species of sheep and goats (Mycoplasma agalactiae) that is highly cross-reactive with M. bovis. Three of the MAbs were used on Western blots of trypsin-treated whole organisms to determine if the antigens were exposed on the surface of M. bovis By isotyping, MAbs were identified as kappa chain IgG1 (3 MAbs), and IgM (3MAbs). The MAbs reacted with all six M. bovis strains in IFA, ELISA, and Western blots. Four of the antigens recognized were highly specific for M. bovis in ELISA, and 3 were cross-reactive with M. agalactiae or other bovine mycoplasmas in Western blots. One MAb reacted with multiple bands with all M. bovis strains, indicating recognition of a size-variant antigen. The size-variant antigen and one of the M. bovis-specific antigens were recognized as surface proteins. A large M. bovis-specific antigen was a conserved cytosolic protein. The M. bovis antigens discovered may be used for specific detection of the organism or measurement of antibody responses, particularly if used in tests with nondenatured alkali-treated antigen, such as ELISA.
牛支原体是牛乳腺炎、关节炎和肺炎的病原体。制备了六种抗牛支原体的单克隆抗体（MAb），并用于鉴定该支原体的特异性抗原。通过间接荧光抗体试验（IFA）、酶联免疫吸附测定（ELISA）和免疫印迹法检测了单克隆抗体对六种牛支原体菌株的反应性。通过相同方法针对其他8种牛支原体和1种与牛支原体高度交叉反应的绵羊和山羊支原体（无乳支原体）检测了这些单克隆抗体的特异性。三种单克隆抗体用于经胰蛋白酶处理的全菌体的蛋白质免疫印迹分析，以确定抗原是否暴露于牛支原体表面。通过血清分型，单克隆抗体被鉴定为κ链IgG1（3种单克隆抗体）和IgM（3种单克隆抗体）。这些单克隆抗体在IFA、ELISA和蛋白质免疫印迹中与所有六种牛支原体菌株发生反应。在ELISA中识别出的四种抗原对牛支原体具有高度特异性，在蛋白质免疫印迹中，有三种与无乳支原体或其他牛支原体发生交叉反应。一种单克隆抗体与所有牛支原体菌株的多条条带发生反应，表明识别出一种大小可变的抗原。大小可变抗原和一种牛支原体特异性抗原被识别为表面蛋白。一种大的牛支原体特异性抗原是一种保守的胞质蛋白。发现的牛支原体抗原可用于该病原体的特异性检测或抗体反应的测定，特别是如果用于非变性碱处理抗原的检测，如ELISA。
