Healey M C, Hwang H H, Kleinschuster S J, Johnston A V, Symons K S
Department of Animal, Dairy, and Veterinary Sciences, College of Agriculture, Utah State University, Logan 84322.
Am J Vet Res. 1988 Nov;49(11):1824-31.
Three monoclonal antibodies (LG17, LG30, LG33) were used in the indirect fluorescent antibody test, the ELISA, and the immunoelectrotransfer blot technique to identify group-specific and strain-specific epitopes on the outer membranes of Actinobacillus seminis, A actinomycetemcomitans, and 17 field isolates of Actinobacillus spp. The field isolates had been obtained by bacteriologic culture of specimens from ram lambs with epididymitis. Only antibody LG33 consistently had specificity for an outer membrane epitope shared by most of the bacterial isolates tested. Staining of polyacrylamide gels with periodic acid-Schiff reagent, Sudan black B, and Coomassie brilliant blue R250 indicated that target antigens for antibodies LG17 and LG33 contained carbohydrate and lipoprotein components, respectively. The chemical composition of the LG30 target antigen was not determined because of its instability after exposure to sodium dodecyl sulfate. Discontinuous-gradient polyacrylamide gel electrophoresis in sodium dodecyl sulfate and spectrophotometric scans of the gels were used to analyze n-octyl-beta-D-glucopyranoside protein extracts from A seminis, A actinomycetemcomitans, and 13 representative field isolates of Actinobacillus spp. Bacterial isolates could be grouped according to their protein profiles. The first group consisted of A seminis, A actinomycetemcomitans, and 7 field isolates of Actinobacillus spp, all of which shared common protein bands with molecular masses of approximately 94 kilodaltons (kD), 64 kD, 60 kD, 52 kD, 44 kD, and 26 kD. The second group was composed of 6 field isolates, each with unique protein profiles; isolates had relatively few protein bands in common. These data suggested that members of the genus Actinobacillus cultured from ram lambs with epididymitis probably include a number of various strains.
使用三种单克隆抗体（LG17、LG30、LG33）进行间接荧光抗体试验、酶联免疫吸附测定（ELISA）和免疫电转移印迹技术，以鉴定精液放线杆菌、伴放线放线杆菌以及17株放线杆菌属野外分离株外膜上的群特异性和菌株特异性表位。这些野外分离株是通过对患附睾炎的公绵羊羔的标本进行细菌培养获得的。只有抗体LG33始终对大多数测试细菌分离株共有的一种外膜表位具有特异性。用高碘酸-希夫试剂、苏丹黑B和考马斯亮蓝R250对聚丙烯酰胺凝胶进行染色表明，抗体LG17和LG33的靶抗原分别含有碳水化合物和脂蛋白成分。由于LG30靶抗原在暴露于十二烷基硫酸钠后不稳定，因此未确定其化学成分。使用十二烷基硫酸钠中的不连续梯度聚丙烯酰胺凝胶电泳和凝胶的分光光度扫描来分析精液放线杆菌、伴放线放线杆菌以及13株代表性放线杆菌属野外分离株的正辛基-β-D-吡喃葡萄糖苷蛋白提取物。细菌分离株可根据其蛋白质谱进行分组。第一组包括精液放线杆菌、伴放线放线杆菌以及7株放线杆菌属野外分离株，所有这些分离株都有分子量约为94千道尔顿（kD）、64 kD、60 kD、52 kD、44 kD和26 kD的共同蛋白条带。第二组由6株野外分离株组成，每株都有独特的蛋白质谱；分离株之间共同的蛋白条带相对较少。这些数据表明，从患附睾炎的公绵羊羔中培养出的放线杆菌属成员可能包括许多不同的菌株。
