Monoclonal Antibody Competitive Elisa For Rapid Detection Of The Establishment Of Salmonella Enteritidis Method

Establishment of A Monoclonal Antibody ased CLISA to Detect S. enteritidis Abstract:In this present study, a monoclonal antibody(McAb) 347-26 specific to 09,was used to establish competitive Enzyme-linked immunosorbed assays(C-ELISA) of detection for Salmonella enteritidis. A hybridomas cell strain 3-47-26 was screened and preparated the high valence monoclonal antibody. The linking method of McAb with HRP was improved and the specificity of HRP-3-47-26 was accredited. During the the experiment, Samonella enteritidis Lipopolysaccharidel(LPS) was preparated and was determined the ingredient pertain, 0.1 7mg/ml;Polysaccharide, 136.6ug/ml. The concentration of coating of antigen LPS and HRP-3-47-26 was determined HRP-3-47-26, 1:1 00;LPS,400ng/ml. Stable coating and minimal false positives were achieved by conjugating LPSto poly-L-lysine, which solved the desorption problem. S.enteritidis LPS in samples competed with S.enteritidis LPS coated on microtitre plate, and competition first reduced binding of McAb to the S.enteritidis LPS on the plate hence to reduce the chromogenic signal, which the aim of detection for sample. The C-ELISA was very specific and no cross reaction, and it was observed in detection sera induced by related Sammonella A-F, Compared with routine detective method for the detection of S.enteritidis, and the sensitivity and specificity of the C-ELISA were 98.7% and 99.3% respectively. A C-ELISA developed was attempted to provide a raped, sensitive and specific test for detecting S.enteritidis. As above study, The program of surveillance and standardized in order to make pave for ELISA kit for the detection of S.enteritidis from different specimens in this study.