Caputa A C, Murtaugh M P, Bey R F, Loken K I
Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Minnesota, St. Paul 55108.
J Clin Microbiol. 1991 Nov;29(11):2418-23. doi: 10.1128/jcm.29.11.2418-2423.1991.
EcoRI-digested DNA from Borrelia burgdorferi was ligated into the dephosphorylated vector pWR590 and transformed into Escherichia coli DH5 alpha. When the gene library was screened, 20 clones reacted with pooled dog sera with high titers (immunofluorescent antibody titer, greater than or equal to 1,280) to this spirochete. One clone expressed a 110-kDa antigen that reacted strongly with the high-titered pooled sera from dogs with Lyme borreliosis and serum from goats immunized with B. burgdorferi. The 110-kDa protein was serum from goats immunized with B. burgdorferi. The 110-kDa protein was expressed with and without isopropyl-beta-D-thiogalactosidase, indicating the protein is not a fusion protein with beta-galactosidase. Monospecific antisera to the 110-kDa antigen recognized a 75-kDa Borrelia protein. Of the sera that reacted with B. burgdorferi by immunoblotting; 57, 100, and 83% of human, dog, and horse serum samples, respectively, reacted with the 110-kDa protein. Sera from individuals that tested negative with a B. burgdorferi lysate with immunoblotting showed no reaction with the 110-kDa protein. The 110-kDa antigen appears to be useful for the diagnosis of Lyme borreliosis.
用EcoRI消化的伯氏疏螺旋体DNA被连接到去磷酸化载体pWR590中,并转化到大肠杆菌DH5α中。当筛选基因文库时,20个克隆与针对该螺旋体的高滴度(免疫荧光抗体滴度大于或等于1280)混合犬血清发生反应。一个克隆表达了一种110 kDa的抗原,该抗原与莱姆病犬的高滴度混合血清以及用伯氏疏螺旋体免疫的山羊血清发生强烈反应。110 kDa蛋白在用和不用异丙基-β-D-硫代半乳糖苷的情况下均有表达,这表明该蛋白不是与β-半乳糖苷酶的融合蛋白。针对110 kDa抗原的单特异性抗血清识别出一种75 kDa的伯氏疏螺旋体蛋白。通过免疫印迹与伯氏疏螺旋体发生反应的血清中,分别有57%、100%和83%的人、犬和马血清样本与110 kDa蛋白发生反应。在用伯氏疏螺旋体裂解物进行免疫印迹检测呈阴性的个体血清与110 kDa蛋白无反应。110 kDa抗原似乎对莱姆病的诊断有用。