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抗B族链球菌碳水化合物单克隆抗体:在乳胶凝集试验和免疫沉淀试验中的应用

Monoclonal antibody to streptococcal group B carbohydrate: applications in latex agglutination and immunoprecipitin assays.

作者信息

Ruch F E, Smith L

出版信息

J Clin Microbiol. 1982 Jul;16(1):145-52. doi: 10.1128/jcm.16.1.145-152.1982.

Abstract

Two monoclonal mouse antibodies with specificities for group B streptococcal capsular antigens were evaluated in assays for the identification of group B streptococci (GBS). One of these antibodies (A9) was shown to precipitate group B carbohydrate antigen in reactions with both purified group B antigen and antigen present in autoclave or enzyme extracts of GBS. A9 antibody was also specific for group B antigen in gel diffusion reactions with extracts of Lancefield group A, B, C, D, F, and G streptococci and was a highly sensitive reagent in detecting soluble group B antigen by counterimmunoelectrophoresis. Antigen extracted from all five serotypes of GBS was shown to be precipitated by A9 antibody. A second monoclonal antibody (C8) was reactive with intact GBS but did not precipitate soluble antigen in bacterial extracts. In contrast with what has been shown for polyclonal rabbit anti-group B antiserum, neither antibody was significantly inhibited in binding or precipitation assays by high concentrations of free rhamnose or other monosaccharides of carbohydrates found in group B antigen. Rhamnose, the most abundant carbohydrate of the group B antigen, does not appear therefore to be an immunodominant determinant in the binding of A9 or C8 antibody. The epitopes of both monoclonal antibodies are exposed on the surface of live as well as heat-fixed GBS cells. A9 antibody-coated latex particles were compared with a commercially available polyclonal latex agglutination reagent and shown to be equally sensitive and specific in the detection of soluble group B antigen in urine and cerebrospinal fluid from patients with GBS infections. Because of its uniformity and defined antigen specificity, which includes reactivity with all five serotypes of GBS, A9 antibody offers the potential of an improved immunodiagnostic reagent for the identification of GBS.

摘要

对两种针对B群链球菌荚膜抗原具有特异性的单克隆小鼠抗体进行了评估,用于B群链球菌(GBS)的鉴定试验。其中一种抗体(A9)在与纯化的B群抗原以及GBS高压灭菌或酶提取物中存在的抗原反应时,能沉淀B群碳水化合物抗原。在与兰氏A、B、C、D、F和G群链球菌提取物的凝胶扩散反应中,A9抗体对B群抗原也具有特异性,并且在通过对流免疫电泳检测可溶性B群抗原时是一种高度敏感的试剂。从GBS的所有五种血清型中提取的抗原均显示可被A9抗体沉淀。第二种单克隆抗体(C8)与完整的GBS有反应,但不能沉淀细菌提取物中的可溶性抗原。与多克隆兔抗B群抗血清的情况不同,在结合或沉淀试验中,高浓度的游离鼠李糖或B群抗原中发现的其他碳水化合物单糖均不会显著抑制这两种抗体。因此,鼠李糖作为B群抗原中含量最丰富的碳水化合物,似乎不是A9或C8抗体结合中的免疫显性决定簇。两种单克隆抗体的表位在活的以及热固定的GBS细胞表面均有暴露。将A9抗体包被的乳胶颗粒与市售的多克隆乳胶凝集试剂进行比较,结果显示在检测GBS感染患者尿液和脑脊液中的可溶性B群抗原时,二者具有同等的敏感性和特异性。由于A9抗体具有均一性和明确的抗原特异性,包括与GBS的所有五种血清型都有反应,因此它有望成为一种改进的免疫诊断试剂,用于GBS的鉴定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78b7/272311/4fe22c3482b8/jcm00144-0173-a.jpg

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