|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)
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.