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Development of rapid detection method of viable Escherichia coli and coli-form

Research Project

Project/Area Number 07660168
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 食品科学・栄養科学
Research InstitutionKYUSHU UNIVERSITY

Principal Investigator

MIYAMOTO Takahisa  Kyushu Univ.Food Sci.& Technol. Associate Prof., 農学部, 助教授 (70190816)

Co-Investigator(Kenkyū-buntansha) HONJOH Ken-ichi  Kyushu Univ.Food Sci.& Technol. Assistant Prof., 農学部, 助手 (00264101)
HATANO Shoji  Kyushu Univ.Food Sci.& Technol. Full Prof., 農学部, 教授 (30038260)
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1996: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1995: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsEscherichia coli / coli-form / monoclonal antibody / Rapid detection
Research Abstract

Three anti-E.coli monoclonal antibodies were prepared. Among them, MAb-C was most specific to E.coli. The MAb did not react with other bacteria such as Enterobacter aerogenes, Klebsiella pneumoniae, Proteus vulgaris, Serratia marcescens, and Salmonella typhimurium. As a preliminary experiment to develop rapid detection of viable cells, ATP assay was performed on E.coli harvested on a membrane filter. By measuring light emission, about 10^4 CFU of E.coli was detected. The same membrane was treated with peroxidase-labeled MAb-C and light emission was measured after reaction with luminol solution (MAb assay). About 10^3 CFU of E.coli were detected with the Mab assay. When the membrane was incubated in Nutrient broth for 6 hr at 35゚C , E.coli was detected at 1 CFU by both methods. Initial cell nember and total light emission counts or total light emission points on the filter was strongly correlated. Correlation coefficients were calculated to be 0.95 and 0.89, respectively, for total light emission counts and total light emission points. In the case of MAb assay, correlation was not so strong between initial cell number and total light emission points (r=0,76). Further work is under way to obtain more specific MAb to E.coli.

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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