| Project/Area Number |
09460146
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied veterinary science
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| Research Institution | Coolage of Agriculture, Osaka Prefecture University |
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Principal Investigator |
UEMURA Takashi Coll.of Agriculture, Osaka Prefecture University, Professor, 農学部, 教授 (40081591)
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| Co-Investigator(Kenkyū-buntansha) |
KANEKO Kenichi College of Agriculture, Tokyo University of Agriculture and Technology, Associat, 農学部, 助教授 (30109508)
ITOH Kikuji Graduate School of Agriculture and Biological Sciences, Tokyo University, Associ, 大学院, 助教授 (50100045)
SUGII Shunji Coll.of Agriculture, Osaka Prefecture University, Professor, 農学部, 教授 (70162865)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥8,700,000 (Direct Cost: ¥8,700,000)
Fiscal Year 1998: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1997: ¥6,000,000 (Direct Cost: ¥6,000,000)
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| Keywords | Escherichia coli / Clostridium perfringens / enterotoxin / O157 / ELISA / flow cytometry / ウェルシュ菌 / 病原性大腸菌 / カンピロバクター / ウェルシュ菌エンテロトキシン / 0157 |
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| Research Abstract |
(1) To develop a rapid and specific method to detect and/or identify enterohemorrhagic Escberichia coli O 157 : H7, two mouse monoclonal antibodies (MCA) were prepared. Specificities of these two MAbs (1D9 and 3E8) were determined by flow cytometry method (FCM). 3E8 and 1D9 were found to react with E.coli O157 : H7, Citrobacter freundii and Salmonella group N (O : 30), but not with E bermannii With a mixture containing strains of E coil O157 : H7 and E coli O6 : Hl, two different peaks appeared in FCM with MCA, whereas a single peak appeared with polyclonal rabbit antiserum. From these findings, FCM with MCA is suggested to be a rapid, specific, and useful method to detect and identify strain(s) of E celi O157 : H7 in food ingredients (Ref.#1).
(2) FCM with fluorescent-labeled anti-CPE antibody was applied to develop a rapid, specific, and convenient method to detect enterotoxin (CPE) exposed on the surface of spores of Clostridium perfringens. The results obtained indicate that FCM can specifically detect CPE exposed on C perfringens spores for a short time. Thus, FCM is found to be a rapid, specific, and convenient assay method for detection of CPE exposed on C perfringens spores, suggesting that it will be hopefully useful to diagnose food poisoning caused by C perfringens (Ref.#2).
(3) Sandwich enzyme-linked immunosorbent assay (ELISA) was developed for the quantitative estimation of C perfringens enterotoxin (CPE) using* monoclonal* and* polyclonal* antibodies* as* capturing* and* detecting* antibodies* respectively.* The dose- dependent relationship between absorbance at 405 nm and concentration of purified CPE was obtained over the range of 0.5 ng/ml-8.0 ? g/ml. The sandwich ELISA is found to detect crude CPE in culture and CPE in 10% fecal extracts. This method is a convenient, rapid and sensitive for specific detection of GPE(Ref#3).
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