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Simple and Rapid Identification of the Mutans Group of Streptococci by Enzyme Immunoassay using Monoclonal Antibodies

使用单克隆抗体通过酶免疫分析简单快速地鉴定链球菌变形菌群

基本信息

批准号：
04671266
负责人：
HIROTA Katsuhiko
金额：
$ 1.15万
依托单位：
The University of Tokushima
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1992
资助国家：
日本
起止时间：
1992 至 1993
项目状态：
已结题

项目摘要

Five monoclonal antibodies (MAbs) with defferent specificity were tested in enzyme immunoassay (EIA) for their reactivity with reference strains of oral bacteria. The MAbs, a-21, b3-22, f-89, s3-9, h-448 reacted specifically with S.cricetus (serotype a), S.rattus (serotype b), S.mutans (serotypes c, e and f), S.mutans (serotype e), and S.sobrinus (serotypes d and g), restectively.The results indicate a diversity of biochemical properties in clinical isolates and suggest that the MAbs in enzyme immunoassay be useful for rapid and simple identification of the mutans group of streptococci, as compared with the conventional method using biochemical reactions.Based on the results above mentioned the enzyme immunoassay system with monoclonal antibody f-89 was considered to become, after some improvement, very useful as a caries activity test.A monoclonal antibody (MAb f-77) was prepared by fusing spleen cells of immune mice and mouse myeloma cells (Sp2/0-Ag14). The monoclonal antibody reaced in enzyme immunoassay with whole cells of Streptococcus mutans strains (serotypes c, e and f) and whole cells of a reference strain (STCC 19615) and several isolates of Streptococcus pyogenes from patients with acute glomerulonephritis. It was demonstrated in competitive enzyme immunoassay using various haptens that reactions between the MAb f-77 and S.pyogenes cells were completely inhibited by N-acetylglucosamine. It was also shown by immunoelectron microscopy that immunogold particles were observed on the cell surfaces of these streptococcal species.13EA05 : These results indicate that S.mutans and S.pyogenes share an antigen structure with an epitope of N-acetylglucosamine. It is possible that S.mutans may produce the same immune response in humans as dose S.pyogenes and play an important role in some disease processes.

用酶免疫法（EIA）检测了5株不同特异性的单克隆抗体（MAb）与口腔细菌参考菌株的反应性。单克隆抗体a-21、b3-22、f-89、s3-9、h-448与小沟链霉菌发生特异性反应（血清型a），鼠链球菌（血清型B），变形链球菌（血清型c、e和f），变异链球菌（血清型e）和远缘链球菌（血清型D和G），结果表明，临床分离株的生化特性多样，提示酶免疫单克隆抗体可用于快速检测临床分离株的生化特性。与传统的生化反应法相比，本方法简便、快速，可用于链球菌变形菌群的鉴定。根据上述结果，认为单克隆抗体F-89酶免疫分析系统经改进后，作为龋齿活性测试非常有用。通过将免疫小鼠的脾细胞与小鼠骨髓瘤细胞（Sp2/0-Ag 14）融合来制备（MAb f-77）。用该单克隆抗体与变形链球菌C、E、F血清型全细胞、标准株（STCC 19615）全细胞及急性肾小球肾炎患者化脓性链球菌分离株进行酶免疫反应。在使用各种半抗原的竞争性酶免疫测定中证明，MAb f-77和化脓性链球菌细胞之间的反应完全被N-乙酰葡糖胺抑制。免疫电子显微镜还显示，在这些链球菌种的细胞表面上观察到免疫金颗粒。13 EA 05：这些结果表明，变形链球菌和化脓链球菌具有相同的抗原结构，具有N-乙酰葡糖胺表位。变形链球菌可能在人体内产生与化脓性链球菌相同的免疫应答，并在某些疾病过程中发挥重要作用。