| Aeromonas hydropphila is widely distributed in nature, which is commonly found in fresh water, sewage, sludge and human excrement and can infect such bacteria pathogens as wild fish, shrimp, frog and turtles by means of damaged skin and the infected wound and even enter the food chain to infect human beings. Aeromonas hydrophila is characterized with wide prevalence, fast incidence and high death rate, causing great economic loss to fishery and threatening the development of fishery and other aquaculture industries. Currently , the researches on Aeromonas hydrophila both home and abroad are limited to the study on pathogenic microorganism, and the diagnosis of this disease mainly depend on pathological observation and bacteriological observation, and the accuracy and detection speed of both methods are limited. With regard to prevention and treatment, antibiotics or the inactivated vaccine is still the major product, whose safety and efficacy are far from being satisfactory. In view of this, this research develops anti-idiotype monoclonal antibody (AId mAbs) against Aeromonas hydrophila to provide new technological methods for the prevention of Aeromonas hydrophila and provide reference for the development of other fish vaccine. The specific experiment methods and results were as follows:By immunizing mouse with monoclonal antibody 1G10 against Aeromonas hydrophila made in our lab, cell fusion, screening and cloning.of hybridomas, four anti-idiotype strains of hybridoma were got and named 1F9,1H10,3F5 and 4G3 respectively. The titers of these supernatant and ascites were 1×10-1～1×10-2and 1×10-4～1×10-5 respectively. The secreted IgG of the 1F9,1H10,4G3 hybridoma cells are IgG2b;The secreted IgG of the 3F5 hybridoma cell is IgG3. The results showed that the reaction of four anti-idiotype mAb and Zelanian rabbit antiserum against Aeromonas hydrophila could be inhibited by Aeromonas hydrophila at different levels. The results of ELISA competitive experiment shows that four anti-idiotypic antibody and multi-clonal anti-body can be inhibited by Aeromonas hydrophila and the inhibition rate increases with the increase of the concentration of Aeromonas hydrophila bacteria. It indicates that anti-idiotypic monoclonal antibody competes with Aeromonas hydrophila。The results show that 1F9,1H10 have higher inhibition rate than the other two, suggesting that there are analog to Aeromonas hydrophila bacteria and have the best antigenicity. The identification result of biological functions shows that the BALB/c mouse serum for preparing 4 Aeromonas hydrophila strains anti-idiotypic monoclonal antibody ascites can have some reactions with Aeromonas hydrophila and the titer was 1:100 to 1:1000. It shows that the serum of mice contain anti-Aeromonas hydrophila antibody, suggesting that 4 Aeromonas hydrophila anti-idiotypic monoclonal antibodies have the role of simulating Aeromonas hydrophila and can be used as vaccine for the prevention of fish Aeromonas hydrophila.In conclusion, by using hybridoma technique, anti-idiotypic monoclonal antibody against Aeromonas hydrophila are developed to provide certain condition fo the prevention of Aeromonas hydrophila. After retrieval, it is found that so far there has no been literature and patent report on the successful research and development of anti-idiotypic antibody vaccines against Aeromonas hydrophila.
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