1985 Fiscal Year Final Research Report Summary
Development of an assay kit for heat-stable enterotoxin (ST) produced by enterotoxigenic Escherichia coli.
| Project/Area Number |
58870033
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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 |
細菌学
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| Research Institution | The University of Tokyo (1985)
Osaka University (1983-1984) |
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Principal Investigator |
TAKEDA Y. 東京大学, 医科研, 教授 (30029772)
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| Project Period (FY) |
1983 – 1985
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| Keywords | Enterotoxigenic Escherichia coli / Heat-stable enterotoxin / STh / STp / Synthetic ST / Monoclonal antibody / competitive ELISA |
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| Research Abstract |
Enterotoxigenic Escherichia coli produces two distinct diarrheagenic enterotoxins. One is heat-labile enterotoxins (LT) which is inactivated by heating at 60゜C for 10 minutes and the other is heat-stable enterotoxin (ST) which is not inactivated by heating at 100゜C for 10 minutes. To detect LT, several simple assay methods have been developed and used in routine assays. For ST, however, suckling mouse assay is an only available practical method.
To develop a simple assay to detect both STh and STp was a major objective of this study and followeing results were obtained.
1. Primary amino acid sequences of both STh and STp were determined.
2. Both STh and STp were chemically synthesized and it was found that the cemically synthesized .ST's were biologically and immunologically active.
3. Several short analogues of STh and STp were chemically synthesized and their biological and immunological activities were examined. It was found that an essential structure for ST toxicity resided on a peptide consisted of 13 amino acids.
4. A monoclonal antibody against STh that has a neutralizing activity of STh and STp activity was prepared.
5. A competitive ELISA system by using a synthesized ST as a solid phase and the monoclonal antibody prepared was developed. Sensitivity of the ELISA developed was about 1 ng/ml.
6. A kit to assay ST-producing enterotoxigenic Escherichia coli was developed and it was demonstrated that the developed kit was useful for a routine assay.
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