权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Development of diagnostic system in periodontal disease using synthetic peptides containing epitopes of surface protein of periodontopathic bacteria

使用含有牙周病细菌表面蛋白表位的合成肽开发牙周病诊断系统

基本信息

批准号：
07557114
负责人：
OGAWA Tomohiko
金额：
$ 3.71万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07557114/
关键词：
Periodontal disease Porphyromonas gingivalis Fimbriae Epitope of B cell Serum Diagnosis Antibody Paper point Porphyromonas gingivalis

项目摘要

Porphyromonas gingivalis is an anaerobic gram-negative rod which has been isolated frequently from the periodontal pockets of patients with periodontal disease. The organism has been implicated as a major pathogen in the development of adult periodontitis. The ELISA testing method with a cellulose acetate paper point for detecting P.gingivalis (P.g.) and its antibody responses in subgingival plaque samples were investigated. Using multipin peptide synthesis technology, several sequential overlapping peptides were synthesized. Six and seven immunodominant regions within 41K-fimbriae and 72-CSK protein molecule, which definitely reacted with the serum of patients with periodontal diseases, were detected, respectively. Rabbit polyclonal IgG (Poly Pgf-I) and mouse monoclonal antibodies (mAbs Pgf-I) against 41K-fimbriae were obtained. Poly Pgf-I and mAbs Pgf-I reacted with cell suspensions consisting of the range of 20,000 to 2,000,000 P.g.cells by the ELISA method. The developed paper point could be classified into four degrees -, +, ++ and +++ according to the number of P.g.cells. Poly Pgf-I and mAbs Pgf-I reacted with P.g.cells, while neither of these antibodies reacted with whole cells of other oral or nonoral black-pigmented bacterial species. Analysis by the ELISA method resulted in definite immunoreactivity in subgingival plaque samples of patients with adult periodontitis, while no detectable reactivity was found in those of normal healthy subjects. Among the patients, high titers of both 41K-fimbria-and 72K-CSP-antibodies or against each protein antigens separately were observed in the serum specimens tested. The 41K-fimbriae-specific IgG subclass responses were elevated in the advanced stage of adult periodontitis, and IgG4 was dominant. The simple and rapid ELISA method using the paper point may be useful for the identification of P.g. and detection of P.g.-specific antibodies.

牙龈卟啉单胞菌是一种厌氧革兰氏阴性杆菌，经常从牙周病患者的牙周袋中分离出来。该生物体被认为是成人牙周炎发展的主要病原体。研究了用醋酸纤维素纸尖检测牙龈卟啉单胞菌（P.g.）的ELISA测试方法及其在龈下菌斑样本中的抗体反应。使用多针肽合成技术，合成了几个连续重叠的肽。 41K-菌毛和72-CSK蛋白分子内分别检测到6个和7个与牙周病患者血清发生明显反应的免疫显性区域。获得了针对 41K 菌毛的兔多克隆 IgG (Poly Pgf-I) 和小鼠单克隆抗体 (mAbs Pgf-I)。通过 ELISA 方法，聚 Pgf-I 和单克隆抗体 Pgf-I 与由 20,000 至 2,000,000 个 P.g. 细胞组成的细胞悬液发生反应。所开发的纸点根据P.g.细胞的数量可分为-、+、++、+++四个等级。聚 Pgf-I 和单克隆抗体 Pgf-I 与 P.g. 细胞发生反应，而这些抗体均不与其他口腔或非口腔黑色素细菌物种的完整细胞发生反应。 ELISA方法分析表明，成人牙周炎患者的龈下菌斑样本中有明确的免疫反应性，而在正常健康受试者的龈下菌斑样本中未发现可检测到的反应性。在患者中，在测试的血清样本中观察到高滴度的41K-菌毛和72K-CSP-抗体或分别针对每种蛋白抗原的抗体。 41K菌毛特异性IgG亚类反应在成人牙周炎晚期阶段升高，且以IgG4为主。使用纸点的简单快速的 ELISA 方法可能有助于 P.g. 的鉴定。以及 P.g. 特异性抗体的检测。