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猪痢疾短螺旋体脂寡糖特异性单克隆抗体的制备与鉴定

Production and characterization of monoclonal antibodies specific for lipooligosaccharide of Serpulina hyodysenteriae.

作者信息

Westerman R B, Phillips R M, Joens L A

机构信息

Department of Veterinary Diagnostic Investigation, College of Veterinary Medicine, Kansas State University, Manhattan 66506, USA.

出版信息

J Clin Microbiol. 1995 Aug;33(8):2145-9. doi: 10.1128/jcm.33.8.2145-2149.1995.

Abstract

Serpulina (Treponema) hyodysenteriae is the causative agent of swine dysentery, a contagious mucohemorrhagic disease of the colon. Diagnosis of swine dysentery is extremely difficult because of the presence of cross-reactive antibodies to the proteins of S. hyodysenteriae and Serpulina innocens, a nonpathogenic inhabitant of the porcine large intestine. Therefore, monoclonal antibodies (MAbs) against the serotype-specific lipooligosaccharide (LOS) antigens of S. hyodysenteriae were produced to rapidly differentiate S. hyodysenteriae from S. innocens. Whole-cell preparations of S. hyodysenteriae serotypes 1 through 7 were used as antigens. MAbs were characterized by an indirect enzyme-linked immunosorbent assay with whole-cell or LOS antigen and by Western blot (immunoblot) analysis with whole-cell lysates as antigen. A total of 12 LOS-specific MAbs which could identify and differentiate the seven original serotypes of S. hyodysenteriae were produced. The MAb serospecificities are as follows: MAb 9G8, serotype 1; MAb 31D9, serotype 2; MAb 7D3, serotypes 2 and 7; MAb 24B7, serotype 3; MAb 13C2, serotype 4; MAb 18E9, serotype 4; MAb 2B7, serotype 6; MAb 1D2, serotypes 2, 5, and 7; MAb 9C5, serotypes 2, 5, and 7; MAb 11C9, serotype 7; MAb 11E10, serotype 7; and MAb 6G11, serotype 7.

摘要

猪痢疾蛇形螺旋体(密螺旋体属)是猪痢疾的病原体,猪痢疾是一种结肠的传染性黏液出血性疾病。由于存在针对猪痢疾蛇形螺旋体和无害蛇形螺旋体(猪大肠中的一种非致病性栖居菌)蛋白质的交叉反应抗体,猪痢疾的诊断极其困难。因此,制备了针对猪痢疾蛇形螺旋体血清型特异性脂寡糖(LOS)抗原的单克隆抗体,以快速区分猪痢疾蛇形螺旋体和无害蛇形螺旋体。使用猪痢疾蛇形螺旋体1至7血清型的全细胞制剂作为抗原。通过用全细胞或LOS抗原进行间接酶联免疫吸附测定以及用全细胞裂解物作为抗原进行蛋白质印迹(免疫印迹)分析来鉴定单克隆抗体。共产生了12种LOS特异性单克隆抗体,它们能够识别和区分猪痢疾蛇形螺旋体的7种原始血清型。单克隆抗体的血清特异性如下:单克隆抗体9G8,血清型1;单克隆抗体31D9,血清型2;单克隆抗体7D3,血清型2和7;单克隆抗体24B7,血清型3;单克隆抗体13C2,血清型4;单克隆抗体18E9,血清型4;单克隆抗体2B7,血清型6;单克隆抗体1D2,血清型

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