1987 Fiscal Year Final Research Report Summary
Research on the Deveropment of a practical protein A ELISA for microbiological monitoring and infectious disease diagnosis in laboratory animals
| Project/Area Number |
60880012
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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 |
Laboratory animal science
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| Research Institution | Central Institute for Experimental Animals |
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Principal Investigator |
KAGIYAMA Naoko Central Institute for Experimental Animals, 動物医学研究室, 室長 (50124269)
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| Co-Investigator(Kenkyū-buntansha) |
櫻井 美典 わかもと製薬(株), 生化学研究部, 研究員
SUZUKI Hirokazu Wakamoto Parmaceutical Co. Ltd., 副主任研究員
小林 憲明 北里衛生専門学院, 助手
TERADA Eiji Kitasato University, 衛生学部, 講師 (10113440)
TAKAKURA Akira Central Institute for Experimental Animals, 動物医学研究室, 研究員 (60167484)
KOBAYASHI Noriaki Kitasato University
SAKURAI Yoshinori Wakamoto Parmaceutical Co. Ltd.
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| Project Period (FY) |
1985 – 1987
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| Keywords | Rat / Infectious disease diagnosis / ELISA / Protein A; Sendai Virus / Mouse hepatitis virus / Sendai vitus / mouse hepatitis virus / Mycoplasma pulmonis |
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| Research Abstract |
1) Procedures for preparation of antiogens and conditions of reactions in the ELISA were investigated. Protein concentrations of 5ug/ml for Sendai virus and Mycoplasma pulmonis and 10<micrn>g/ml for each Nu-67 and S strain of MHV, solubilized with carbonate-bicarbonate buffer, pH 9.6, were selected for antigen preparation.
2) A quality assurance system for the antigens was established by SDS-PAGE and the immuno- blotting method.
3) The antigens and reagents prepared for the ELISA Kits Kept their sensitivity and specificity for at least 6 months at 4゜C.
4) The boundary OD492nm value between negative and positive was determined to be 0.3 (2-wavelength microplate photometer: Corona).
5) Three field investigations on the efficacy of the ELISA Kits were performed with the cooperation of a total of 227 organizations.
6) A total of 7,528 serum samples was collectled from the cooperating organizations, and a comparison was performed between the judgements on the same sera by photometer (the research project team) and by naked eye (cooparating organization) observations.
7) It was clarified that the inconsistency between the two judgements was due to the difficulty of thenaked eye to recognize boundary positive reactions.
8) This problem was solved by using the improved antigen preparations as mentioned in 1., with increased chromogenic reactivities.
9) Nonspecific coloring reactions rarely occurred in the sera collected from non-infected animals.
10) Nonspecific weak coloring was observed in the sera of autoimmune mice of such strains as MRL/1pr and NZB/NZW F1, and sentinel mice were considered to be necessary in such cases.
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