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Hydrophila Aeromonas (Aeromonas hydrophila, Ah) is widely distributed in nature, commonly found in freshwater, sewage, sludge, soil and human feces, common bacterial pathogens of farmed and wild fish, shrimp, frogs, turtles and other creatures through damaged skin and wound infections the above biological, and further infect humans through the food chain. The bacteria caused fish hydrophila Aeromonas disease is widely popular fast onset, high mortality, often resulting in huge economic losses for aquaculture fisheries, serious harm to the development of fisheries and aquaculture. At home and abroad for hydrophila Aeromonas fish diseases, limited to pathogenic microbiology, diagnosis depends on pathological observation and bacteriological identification of the disease, the two methods have certain limitations in accuracy and detection speed ; prevention and treatment with antibiotics or deactivation, inactivated vaccine safety and efficacy are not ideal. In view of this, the study design preparation addicted to water Aeromonas anti-idiotypic monoclonal antibody, a new technology for disease detection and treatment of Aeromonas hydrophila, and the vaccine provides reference for other fish. Specific experimental method and the results are as follows: Aeromonas hydrophila in this room homemade immune and active anti-monoclonal antibody 1G10, through animal immunization, cell fusion, screening and cloning, 4 secreting anti-idiotype antibody The hybridoma cell line, were named as 1F9, 1H10, 3F5 and 4G3. The culture supernatant of the hybridoma cell line and BALB / c mouse ascites anti-idiotype antibody titer range of 1 × 10 -1 ~ 1 × 10 -2 1 × 10 -4 ~~ 1 × 10 -5 . Four hybridoma cell lines secreted IgG subclasses: 1F9, 1H10, 4G3 IgG2b; 3F5 for IgG3. ELISA competitive inhibition experiments results show: four anti-idiotype antibody and rabbit anti addicted to water Aeromonas polyclonal antibody binding can be hydrophila Aeromonas inhibition, and inhibition percentage as the concentration of Aeromonas hydrophila the increase of anti-idiotype monoclonal antibody 4 hydrophila Aeromonas hydrophila Aeromonas between competing relationship have analog addicted to water Aeromonas antigen features. Percentage inhibition results show 1F9, 1H10 by the degree of inhibition than the remaining two by the high degree of inhibition, suggesting that its analog hydrophila Aeromonas antigen activity. The biological function of identification results showed that the preparation of 4 hydrophila Aeromonas anti-idiotype monoclonal antibody ascites BALB / c mouse serum can Aeromonas hydrophila bacteria reaction, a titer of 1:100 to 1: 1000, indicating that the serum containing anti addicted to water Aeromonas antibody, anti-idiotypic monoclonal antibody analog hydrophila Aeromonas epitopes prompted four hydrophila Aeromonas available for the prevention fish hydrophila Aeromonas bacteria disease vaccine. In summary, this study is the use of the hybridoma technique were prepared polyclonal anti-idiotypic monoclonal antibody resistant Aeromonas hydrophila, Aeromonas hydrophila Aeromonas disease prevention. Upon retrieval, and so far no addicted to water Aeromonas anti-idiotype antibody vaccine successfully prepared literature and patent reports.
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