Preparation And Identification Of The Monoclonal Antibody Against Clostridium Chauvoei

Also known as Blackleg, emphysematous carbuncle is acute, a febrile infectious diseases of ruminant animals which caused by Clostridium chauvoei. The diseases appear sporadic or endemic mainly and widely distribute in nature. On certain conditions, it can cause serious diseases of the body and bring huge financial losses to animal husbandry and aquaculture. And there is no complete detection technology against the diseases. The key of protection against the fungus is check out the pathogen and diagnose disease exactly and quickly. For this reason, this experiment uses normative Clostridium chauvoei C54-1 broken as antigen to immune 6-8 week-old female Balb/c mice, and utilize traditional hybridoma technology, the hybridoma cell lines of Clostridium chauvoei monoclonal antibody secreted stably are obtained and named for 3B7 after screened and cloned by indirect ELISA and limited dilution method. Use indirect ELISA and Western blotting specific identification for the hybridoma cell, make the monoclonal antibody subclass kit identify its MAbsubtypes, and use conventional approach for chromosome analysis and titer evaluation of ascites antibody. The results of specificity test show that the MAb have response with Clostridium chauvoei only, and have no response with Clostridium septique,Cl.perfri-ngens(type A,B,C,D,E),E. coli 0:157, Salmonella 045-2 (D group) and Brucella M5-90 etc, it has stronger specificity. After identification of monoclonal antibody subclass kit, the heavy chain of the MAb was IgA, and k chain belonged to light chain; The antibody titer was 1×105. this experiment supplies basis and new technology method for rapid clinical diagnosis and treatment of Clostridium chauvoei, and laid the foundation for development of colloidal gold dipsticks used for detection.