Novel indirect enzyme-linked immunosorbent assay (ELISA) method to detect Total E. coli in water environment.
In order to establish ELISA (enzyme-linked immunosorbent assay) method to detect Total E. coli in water environment, E. coli multi-characters antigens in water environment were prepared according to the characters of kinds of E. coli serotypes, including antigen of whole cell, antigen of disrupted whole cell, somatic antigen, flagellar antigen and fimbrial antigen. Total E. coli polyclonal antibodies were obtained from the New Zealand rabbits immunized with these five antigens, respectively. Antibodies generated in this research are with high titers and good purity, can conjugate with antigens, specifically, stably and strongly. Indirect ELISA shows the titers of antibody of whole cell and antibody of disrupted whole cell are both over 1x10(5). The cross-reactivity of the antibody is from 12 to 30% which indicate the specificity of the antibody against Total E. coli. Based on these antibodies, we established indirect ELISA method to detect Total E. coli in water environment. The matrix effects were studied and the results show that there is no significant influence by all the factors. The ELISA result shows that the detection limitation could be 10(4) CFU (colony forming units) L(-1). The indirect ELISA method developed in this study is well suited for Total E. coli analysis in real water samples as a rapid screen method.
Wang
Na
N
ESPC State Key Joint Laboratory, Department of Environmental Science and Engineering, Tsinghua University, Beijing 100084, China.
He
Miao
M
Shi
Han-Chang
HC
D000907:Antibodies, Bacterial / Q000032:analysis; D000942:Antigens, Bacterial / Q000032:analysis; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004926:Escherichia coli / Q000302:isolation & purification*; D014871:Water Microbiology
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial