The Preparation Of Monoclonal Antibody For 6 Stains Of Pathogenic E. Coli

E. coli disease is caused by a bacterial pathogenic zoonotic disease. It often sojourn, growth, reproduction in the segment of the small intestine of humans and animals, can damage the small intestine characteristic that intestinal villous atrophy, intestinal necrosis, the formation of ulcers. With the development of intensive farming, the disease is an important cause of bacterial diseases of livestock and poultry breeding industry to aquaculture poses a serious threat and significant economic losses. Due to the abuse of antibiotics, E. Coli can easy to produce drug resistance and the resistance patterns become more and more widely.These drugs produce resistant strains have emerged, and there is a growing trend that the drug effect is not ideal after the onset.At the same time it has brought great difficulties for the preparation of E. coli vaccine because of the complex structure of the E. coli antigens and pathogenic E. coli serotypes of many.Six kinds of pathogenic E. coli Colibacillosis which caused what kind of serotype can be rapidly diagnosed according the preparation of antibody.At the same time it has a preventive and therapeutic purp oses for the disease caused by E. coli with the combination of antibodies and coupling drugs,which will reduce E.coli resistance, but also provides guidance for clinical practice.1.Pathogenic E.coli serotypes ScreeningThe serotype identification of 30 strains of suspected pathogenic Escherichia coli w as carried out in this experiment.there were identified and found twelf pathogenic serotypes of E.coli they are respectively O1、O88、O136、 O169、 O144、0157、 O141、 O、O149、O9、O64、O101。2. Preparation of monoclonal antibodies Six of pathogenic E. ColiThe separated six pathogenic E. Coli that incativated by formaldehyde were administered to mice.Using indirect ELISA to detect the serum titer of mice, cell fusion when the titer reached more than 104.the serum positive rate of immunized mice have reached 100%.Mice that have the highest serum titer were boosted in the three days before cell fusion, take 3 days after the booster immunization spleen cells fused with mouse myeloma cells. The cells of fusion rate reached 96.67% and the positive rate of cells have reached 31.5%. Monoclonal growth and strong positive hybridoma cell activity were screened by indirect ELISA method and the use of limited dilution subcloning at least three times to ensure that the positive rate of 100%. The study was undertaken to obtain hybridoma monoclonal antibodies against five kinds of pathogenic E. Coli (they are O1、O88、O136、O169、O144) after three subclones.the antibody titers have reached 105 by indirect ELISA.