Brooker J D, Stokes B
Department of Animal Sciences, Waite Agricultural Research Institute, Glen Osmond, South Australia.
Appl Environ Microbiol. 1990 Jul;56(7):2193-9. doi: 10.1128/aem.56.7.2193-2199.1990.
Monoclonal antibodies were raised against whole cells of two different strains of Selenomonas ruminantium and tested for specificity and sensitivity in immunofluorescence and enzyme-linked immunosorbent assay procedures. Species-specific and strain-specific antibodies were identified, and reactive antigens were demonstrated in solubilized cell wall extracts of S. ruminantium. A monoclonal antibody-based solid-phase immunoassay was established to quantify S. ruminantium in cultures or samples from the rumen, and this had a sensitivity of 0.01 to 0.02% from 10(7) cells. For at least one strain, the extent of antibody reaction varied depending upon the stage of bacterial growth. Antigen characterization by immunoblotting shows that monoclonal antibodies raised against two different strains of S. ruminantium reacted with the same antigen on each strain. For one strain, an additional antigen reacted with both monoclonal antibodies. In the appropriate assay, these monoclonal antibodies may have advantages over gene probes, both in speed and sensitivity, for bacterial quantification studies.
制备了针对两种不同菌株的反刍月形单胞菌全细胞的单克隆抗体,并在免疫荧光和酶联免疫吸附测定程序中测试了其特异性和敏感性。鉴定出了种特异性和菌株特异性抗体,并在反刍月形单胞菌的可溶性细胞壁提取物中证实了反应性抗原。建立了基于单克隆抗体的固相免疫测定法,以定量瘤胃培养物或样品中的反刍月形单胞菌,该方法对10(7)个细胞的敏感性为0.01%至0.02%。对于至少一个菌株,抗体反应程度因细菌生长阶段而异。通过免疫印迹进行的抗原表征表明,针对两种不同反刍月形单胞菌菌株产生的单克隆抗体与每种菌株上的相同抗原发生反应。对于一个菌株,一种额外的抗原与两种单克隆抗体都发生反应。在适当的测定中,这些单克隆抗体在细菌定量研究的速度和敏感性方面可能比基因探针具有优势。