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Development of simple and rapid method using monoclonal antibodies for identification of periodontal bacteria

开发简单快速的单克隆抗体鉴定牙周细菌的方法

基本信息

批准号：
63480459
负责人：
KOGA Toshihiko
金额：
$ 3.14万
依托单位：
National Institute of Health
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1989
项目状态：
已结题

项目摘要

The chemical structures of serotype-specific polysaccharide antigens from Actinobacillus actinomycetemcomitans were determined. Crude antigens were extracted from whole cells of A actinomycetemcomitans by autoclaving. The extracts were purified by ion-exchange chromatography and gel filtration. The serotype a, b and c antigens were composed of repeating units, -3)-6-deoxy-alpha-D-Tal_p-(1-2)-6-deoxy-alpha-D-Tal_p-(1-, -3)-alpha-D-Fuc_p-(1-2)-alpha-L-Rha_p-(1- and -3)-6-deoxy-alpha-L-Tal_p-(1-2)-6-deoxy-alpha-L-Tal_p-(1-, respectively.Twenty-one hybridomas producing monoclonal antibodies (MAbs) to whole cells of A. actinomycetemcomitans strain Y4 were elaborated. Among these MAbs, eight MAbs (MAb Sl-S8) reacted with the serotype b antigen, three MAbs (MAb Ll-L3) reacted with lipopolysaccharide from strain Y4, and four MAbs (MAb Pi-P4) reacted with cell-surface protein antigens of strain Y4.Latex particles sensitized with these MAbs were allowed to react with whole cells and cell extracts of periodontal bacteria. Latex particles sensitized with MAb L2 coagglutinated with whole cells and culture supernatants of all three serotypes of A. actinomycetemcomitans. On the other hand, latex particles sensitized with MAb S5 produced positive agglutination with whole cells and culture supernatants of serotype b A. actinomycetemcomitans and whole cells of Bacteroides gingivalis, but not with heated (100ﾟC, 3 min) cells of B. gingivalis.The simple and rapid latex agglutination assay using MAbs may be useful for the identification and serotyping of A. actinomycetemcomitans strains.

测定了伴放线放线杆菌（Actinobacillusactinomycetemcomitans）中放线菌型特异性多糖抗原的化学结构。用高压灭菌法从伴放线菌（Actinomycetemcomitans）全细胞中提取抗原。提取物通过离子交换层析和凝胶过滤进行纯化。血清型a、B和c抗原由重复单元组成，-3）-6-脱氧-α-D-Tal_p-（1-2）-6-脱氧-α-D-Tal_p-（1-，-3）-α-D-Fuc_p-（1-2）-α-L-Rha_p-（1-和-3）-6-脱氧-α-L-Tal_p-（1-2）-6-脱氧-α-L-Tal_p-21株产生抗A.对伴放线菌Y 4菌株进行了阐述。其中8株单抗（MAB S1-S8）与血清型B抗原反应，3株单抗（MAB L1-L3）与Y 4菌株脂多糖反应，4株单抗（MAB Pi-P4）与Y 4菌株细胞表面蛋白抗原反应。用MAb L2致敏的乳胶粒与所有三种血清型的A.伴放线菌另一方面，用MA B S5致敏的胶乳颗粒与血清型B A的全细胞和培养上清液产生阳性凝集。放线菌共生菌和牙龈拟杆菌的全细胞，但不与加热（100 ℃，3 min）的B细胞。用单克隆抗体建立的简便、快速的乳胶凝集试验可用于牙龈卟啉单胞菌的鉴定和血清分型。放线菌共生菌株。