Rapid detection of enterotoxigenic <i>Escherichia coli</i> O6 in water by using monoclonal antibody and a photon-counting television camera

作     者：Trevanich, S Miyamoto, T Harada, Y Honjoh, K Hatano, S 

作者机构：Kyushu Univ Fac Agr Higashi Ku Fukuoka 8128581 Japan 

出 版 物：《JOURNAL OF FOOD PROTECTION》 (食品保护杂志)

年 卷 期：2000年第63卷第4期

页      面：534-538页

核心收录：

学科分类：0832[工学-食品科学与工程（可授工学、农学学位）] 08[工学] 0836[工学-生物工程] 

主　　题：抗体 单克隆 抗体特异性 细菌毒素类/分离和提纯 肠毒素类/分离和提纯 酶联免疫吸附测定 大肠杆菌 过滤 免疫酶技术 鲁米诺 光子 电视 水微生物学 

摘      要：Monoclonal antibodies (MAbs) raised against Escherichia coli O6:H16 were screened against: 15 strains of E. coli and 19 non-E. coli bacteria. A MAb-luminescence assay using MAb-5.8, which shows no cross-reactions with non-E, coli bacteria, and a photon-counting television camera were developed for rapid enumeration of E. coli O16:H16 in water. The membrane filter that retained bacteria was boiled for 5 min in a buffer and incubated with biotinylated MAb5.8. Alter incubation with streptavidin-peroxidase conjugate, it was reacted with luminol-based reaction mixture. Luminous image and light intensity of the filter was recorded with a Biocell Counter. Levels of E. coli O6 higher than 7 x 10(3) CFU were detected by the MAb-luminescence assay when E. coli O6 was spotted onto the membrane filter. The sample that contained E, coli O6:H16 was filtered through a membrane filter, and the filter that retained bacteria was incubated on a filter paper soaked with nutrient: broth supplemented with 0.5% NaCl at 37 % for 6 h. The number of light emission points on the filter correlated well with initial E. coli O6:H16 counts within the range of 1 to 3 x 10(2) CFU. The correlation coefficient was 0.89.