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通过酶联免疫吸附测定(ELISA)和免疫印迹分析检测犬血清对伯氏疏螺旋体全细胞或重组抗原的反应性。

Reactivity of dog sera to whole-cell or recombinant antigens of Borrelia burgdorferi by ELISA and immunoblot analysis.

作者信息

Magnarelli Louis A, Levy Steven A, Ijdo Jacob W, Wu Caiyun, Padula Steven J, Fikrig Erol

机构信息

Department of Entomology, Connecticut Agricultural Experiment Station, New Haven, CT 06504, *Durham Veterinary Hospital, 178 Parmelee Hill Road, Durham, CT, †Section of Rheumatology, Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06520 and ‡Division of Rheumatic Diseases, Department of Medicine, University of Connecticut Health Center, Farmington, CT 06030, USA.

出版信息

J Med Microbiol. 2001 Oct;50(10):889-895. doi: 10.1099/0022-1317-50-10-889.

Abstract

Enzyme-linked immunosorbent assays (ELISAs) with separate preparations of 10 purified recombinant antigens of Borrelia burgdorferi sensu stricto were used to test sera from 36 dogs not vaccinated with whole cells of this agent and from five dogs vaccinated with whole-cell B. burgdorferi bacteria. All dogs lived in tick-infested areas of Connecticut and south-eastern New York state, USA. The non-vaccinated dogs had limb or joint disorder, lameness and fever during the period 1984-1991 and had antibodies to B. burgdorferi, as determined by a polyvalent ELISA with whole-cell antigen. In re-analyses of sera for total immunoglobulins in ELISAs with recombinant antigens, reactions were most frequently recorded when outer-surface protein (Osp) F, protein (p)35, p37, p39 and p-41G (a flagellin component) were tested separately. Western immunoblots of a subset of 16 sera, positive by ELISA with whole-cell antigen and representing a range of antibody titres (640-40960), verified immune responses to these or other lysed whole-cell antigens. Sera from vaccinated dogs contained antibodies to OspA, OspB, p22, p37 and p41-G. Therefore, serological reactions to OspF, p35 and p39 were the most important indicators of natural exposure to B. burgdorferi. Serum reactivities to these recombinant antigens in ELISAs can be used to help identify possible natural infections of canine borreliosis in dogs not vaccinated with whole-cell B. burgdorferi and to provide information on the geographic distribution of this bacterium.

摘要

使用分别制备的10种伯氏疏螺旋体狭义亚种纯化重组抗原来进行酶联免疫吸附测定(ELISA),以检测36只未接种该病原体全细胞疫苗的犬和5只接种了全细胞伯氏疏螺旋体细菌的犬的血清。所有犬均生活在美国康涅狄格州和纽约州东南部蜱虫滋生的地区。在1984年至1991年期间,未接种疫苗的犬出现肢体或关节紊乱、跛行和发热症状,并且通过使用全细胞抗原的多价ELISA检测发现它们具有抗伯氏疏螺旋体的抗体。在用重组抗原进行的ELISA中对血清总免疫球蛋白进行重新分析时,当分别检测外表面蛋白(Osp)F、蛋白(p)35、p37、p39和p-41G(一种鞭毛蛋白成分)时,反应最为常见。对16份血清的一个子集进行的蛋白质免疫印迹分析(这些血清通过全细胞抗原ELISA呈阳性,代表一系列抗体滴度(640 - 40960)),验证了对这些或其他裂解全细胞抗原的免疫反应。接种疫苗的犬的血清含有针对OspA、OspB、p22、p37和p41 - G的抗体。因此,对OspF、p35和p39 的血清学反应是自然暴露于伯氏疏螺旋体的最重要指标。在ELISA中血清对这些重组抗原的反应性可用于帮助识别未接种全细胞伯氏疏螺旋体疫苗的犬可能存在的犬疏螺旋体病自然感染,并提供有关该细菌地理分布的信息。

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