1989 Fiscal Year Final Research Report Summary
Development of simple and rapid method using monoclonal antibodies for identification of periodontal bacteria
| Project/Area Number |
63480459
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
小児・社会系歯学
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| Research Institution | National Institute of Health |
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Principal Investigator |
KOGA Toshihiko N.I.H., Dept. Dent. Res., Director, 歯科衛生部, 部長 (10037541)
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| Co-Investigator(Kenkyū-buntansha) |
NISHIHARA Tatsuji N.I.H., Dept. Dent. Res., Research Worker, 歯科衛生部, 研究員 (80192251)
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| Project Period (FY) |
1988 – 1989
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| Keywords | Monoclonal Antibody / Periodontal Bacteria / Latex Agglutination Assay / Serotype |
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| Research Abstract |
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.
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