1989 Fiscal Year Final Research Report Summary
DEVELOPMENT OF LATEX AGGLUTINATION TEST FOR SPECIFIC DETECTION OF BACTEROIDES GINGIVALIS
| Project/Area Number |
63870072
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Conservative dentistry
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| Research Institution | OSAKA UNIVERSITY |
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Principal Investigator |
HAMADA Shigeyuki Osaka University Faculty of Dentistry, Professor, 歯学部, 教授 (60028777)
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| Co-Investigator(Kenkyū-buntansha) |
KIMURA Shigenobu Osaka University Faculty of Dentistry, Lecturer, 歯学部附属病院, 講師 (10177917)
OGAWA Tomohiko Osaka University Faculty of Dentistry, Research Associate, 歯学部, 助手 (80160761)
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| Project Period (FY) |
1988 – 1989
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| Keywords | Periodontal disease / Bacteroides gingivalis / Fimbriae / Latex agglutination / IgG / Antibody |
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| Research Abstract |
The purpose of this study was to develop a latex agglutination test to detect the fimbriae of Bacteroides gingivalis cells. This test can be applied to demonstrate fimbrial antigen of B. gingivalis in dental plaque and gingival crevicular fluid for possible diagnosis of periodontal disease.
The fimbriae were mechanically detached from freshly grown cells of B. gingivalis 381 and purified by a DEAE Sepharose Fast Flow column chromatography. Rabbit polyclonal antibody (pAb) and murine monoclonal antibodies (mAbs) to the fimbriae were prepared and were used for sensitization of latex particles. The isotype of mAb was found to be IgM or IgGl, respectively. Sensitization of latex particles was done by incubating them in serially diluted antibody solutions. The agglutination test was carried out on a slide glass and the results were graded (-, +, ++, +++) according to the promptness and extent of the aggregate formation. The latex particles sensitized with pAb agglutinated all B. gingivalis strains tested, while mAb-sensitized particles agglutinated only 8 out of 11 strains tested. Neither pAb- nor mAb-sensitized latex particles reacted with cells of other Bacteroides species including B. melaninogenicus, B. intermedius, B. endodontalis, B. assacharolyticus and B. buccae. Latex particles sensitized with pAb or mAb were agglutinated upon addition of the fimbrial protein, but not LPS or capsular substance of B. gingivalis 381. Furthermore, it was found that both pAb- and mAb-sensitized latex preparations were equally sensitive in detecting the fimbrial antigen (minimum agglutinating dose of fimbriae, 4 ng per test). Our findings indicate that these antibodies may be useful for the preparation of an immunodiagnostic kit for periodontal disease.
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