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基于单克隆抗体的夹心酶联免疫吸附测定法检测百日咳博德特氏菌丝状血凝素

Monoclonal antibody-based sandwich enzyme-linked immunosorbent assay for detection of Bordetella pertussis filamentous hemagglutinin.

作者信息

Gustafsson B, Askelöf P

机构信息

Department of Vaccine Production, National Bacteriological Laboratory, Stockholm, Sweden.

出版信息

J Clin Microbiol. 1988 Oct;26(10):2077-82. doi: 10.1128/jcm.26.10.2077-2082.1988.

Abstract

Hybrid cell lines producing monoclonal antibodies against Bordetella pertussis filamentous hemagglutinin (FHA) were established. The specificity of the antibodies was ascertained by enzyme-linked immunosorbent assay (ELISA), sandwich ELISA, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by electroblotting. The monoclonal antibody-based sandwich ELISA was developed for detection of B. pertussis FHA. The assay had a detection limit of B. pertussis FHA in concentrations ranging from 7 to 15 ng/ml. The assay was also able to detect whole B. pertussis, Bordetella parapertussis, and Bordetella bronchiseptica bacteria. No cross-reactions were observed with strains of Branhamella catarrhalis, Neisseria meningitidis, Haemophilus influenzae, Klebsiella pneumoniae, Legionella pneumophila, Streptococcus miteor, or Streptococcus pneumoniae. The monoclonal antibodies might be useful for the detection of soluble antigens and whole bacteria in clinical samples and for studies of the immunochemical structure of B. pertussis FHA.

摘要

建立了产生抗百日咳博德特氏菌丝状血凝素(FHA)单克隆抗体的杂交细胞系。通过酶联免疫吸附测定(ELISA)、夹心ELISA以及十二烷基硫酸钠-聚丙烯酰胺凝胶电泳后进行电印迹来确定抗体的特异性。开发了基于单克隆抗体的夹心ELISA用于检测百日咳博德特氏菌FHA。该检测方法对百日咳博德特氏菌FHA的检测限为7至15 ng/ml。该检测方法还能够检测完整的百日咳博德特氏菌、副百日咳博德特氏菌和支气管败血博德特氏菌。未观察到与卡他布兰汉菌、脑膜炎奈瑟菌、流感嗜血杆菌、肺炎克雷伯菌、嗜肺军团菌、微小链球菌或肺炎链球菌菌株发生交叉反应。这些单克隆抗体可能有助于检测临床样本中的可溶性抗原和完整细菌,以及用于百日咳博德特氏菌FHA免疫化学结构的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/876c/266819/408a7dd2e2f3/jcm00082-0183-a.jpg

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