Thorns C J, Sojka M G, Mclaren I M, Dibb-Fuller M
Ministry of Agriculture, Fisheries and Food, Central Veterinary Laboratory, Weybridge, Surrey.
Res Vet Sci. 1992 Nov;53(3):300-8. doi: 10.1016/0034-5288(92)90130-t.
A panel of 13 monoclonal antibodies from different hybridomas was produced against a novel salmonella fimbrial antigen expressed predominantly by Salmonella enteritidis strains. The specificity of the monoclonal antibodies to this antigen (SEF14) was confirmed by enzyme-linked immunosorbent assay (ELISA) using purified SEF14, immune electron microscopy and, with 11 monoclonal antibodies, the identification of a repeating protein subunit (14,300kDa) on the antigen. Blocking-ELISA with the monoclonal antibodies identified epitopes in at least three, non-overlapping clusters which appeared evenly distributed on SEF14 in immune electron microscopy. The use of the monoclonal antibodies in direct-binding ELISA on a range of salmonella serotypes suggested that the epitopes on SEF14 are highly conserved and were expressed by all the S enteritidis strains examined; some strains of S dublin and the only strain of S moscow available were the only other serotypes that expressed SEF14. A latex agglutination reagent based on a monoclonal antibody was developed and used to test for SEF14 on 280 strains (representing 120 serotypes in 24 serogroups of salmonellae) that had been grown on Sensitest agar for 18 hours at 37 degrees C. All S enteritidis strains (64) and most S dublin strains (28 of 33) produced SEF14 as did the two strains representing S blegdam and S moscow. SEF14 was not detected in any other strains of serotypes from serogroup D or from any other serogroup examined.
针对一种主要由肠炎沙门氏菌菌株表达的新型沙门氏菌菌毛抗原,制备了一组来自不同杂交瘤的13种单克隆抗体。通过使用纯化的SEF14进行酶联免疫吸附测定(ELISA)、免疫电子显微镜以及利用11种单克隆抗体鉴定抗原上的重复蛋白亚基(14,300kDa),证实了这些单克隆抗体对该抗原(SEF14)的特异性。用单克隆抗体进行的阻断ELISA鉴定出至少三个非重叠簇中的表位,这些表位在免疫电子显微镜下似乎均匀分布在SEF14上。在一系列沙门氏菌血清型的直接结合ELISA中使用这些单克隆抗体表明,SEF14上的表位高度保守,并且在所检测的所有肠炎沙门氏菌菌株中均有表达;都柏林沙门氏菌的一些菌株和仅有的一株莫斯科沙门氏菌是仅有的其他表达SEF14的血清型。开发了一种基于单克隆抗体的乳胶凝集试剂,并用于检测在Sensitest琼脂上于37℃培养18小时的280株菌株(代表沙门氏菌24个血清群中的120个血清型)上的SEF14。所有肠炎沙门氏菌菌株(64株)和大多数都柏林沙门氏菌菌株(33株中的28株)以及代表布莱格丹沙门氏菌和莫斯科沙门氏菌的两株菌株都产生SEF14。在血清群D的任何其他血清型菌株或所检测的任何其他血清群中均未检测到SEF14。