| Project/Area Number |
61571118
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Laboratory medicine
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| Research Institution | Showa University |
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Principal Investigator |
NAKAMURA Yoshiko Showa University Fujigaoka Hospital, 医学部, 助教授 (70119238)
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| Co-Investigator(Kenkyū-buntansha) |
AOKI Yoshio Showa University Fujigaoka Hospital, 医学部, 教授 (50053749)
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| Project Period (FY) |
1986 – 1988
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1988: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1987: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1986: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | Campylobacter jejuni / Monoclonal antibodies / Detection of antigens / Reversed passive latex agglutination / Western blotting / Detection of antibodies / カンピロバクター(Campylobacter jejuni) / モノクローナル抗体 / ウエスタン・ブロット法 / 迅速診断 |
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| Research Abstract |
Campylobacter jejuni, the most frequent pathogens of acute gastroenteritis, usually requires two days for the isolation and identification. We made monoclonal antibodies(MoAb) against C. iejuni so as to search the frequencies of C. jejuni isolation and to establish the method for rapid diagnosis.
1) The frequencies of pathogens isolated from feces were 36% in C. jejuni/ coli, 27% in Klebsiella oxytoca, 14% in enteropathogenic E. coli and 9% in Salmonella spp.
2) Campylobacter coli shared 8% in all C. jejuni/coli.
3) Eleven anti C. jejuni monoclonal antibodies were obtained by means of heat extracted C. jejuni C_1 strain. The epitopes reactive with these MoAb were polypeptides having 45kDa(6 strains) and 65kDa(5 strains) molecular masses on SDS-PAGE. The classes of MoAb were identified as IgM(2 strains) and IgG(9 strains).
4) MoAb(H-194) sensitized latex agglutination(reversed passive latex agglutination, RPLA) was verified to be Campylobacter specific when examined by other pathogens of gastroenteritis.
5) The sensitivity of the RPLA was 4x10^4 CFU/ml, as calculated from the plate-RPLA and colony formation unit of C. jejuni C strain.
6) All clinically isolated 121 strains of C. jejuni and 11 strains of C. coli showed positive RPLA.
7) There was a 96%(249 of 260) total agreement between RPLA and the standard culture method. Among 200 feces showed positive culture, 198 were identified as positive RPLA. Nine feces among 60 negative culture showed positive results.
In summary, the RPLA appears to be useful for rapid detection(within 10 min) of Campylobacter. The method is sensitive, specific, simple to perform and requires no costly instrument.
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