Study On Extraction With Supersonic And Antigenicity Of The Outer-membrane Protein Of Bordetella Avium.

Bordetalla avium disease was a high-contacted infectious disease.In our country,it was discovered by Rui-liang Zhu in 1990.It could arouse that chicken-embryo death;low incubation;chicking acuting death and chicken ophthalmia so on.The infection rate of this disease was 10%～50%,it could bring up illness-chicken grew up lowly, the low redound upon of feedingstuff,and great economy loss.Recent years, the immunity of outer-membrane-protein of bacteria in cell-wall was paid attention to by science workers.the test testified that OMP with well immunity could stimulate not only humoral immunity and cell immunity. We researched the immunity of whole bacterin with Bordetalla avium,the ingredients without immunity were so much that they could influence the immunity,so we began to research extracted craftwork and immunity of outer membrane protein of Bordetella avium, it could help the forward research of subunit vaccine of Bordetella avium.Test one: Development of Indirect ELISA for Detection of Antibodies to Bordetella aviumUsing the Bordetella avium OMP as antigen we developed an indirect enzyme-linked immunosorbent assey (ELISA) for detecting Bordetella avium antibody. The test conditions were optimized by reagent titration utilizing known positive and negative sera. A positive /negative (P/N) value larger than 2.1 was determined as comprehensive positive standard for the ELISA. The performance of the ELISA was assessed for reproducibility and specificity, sensitivity. It was simple, convenient.Test two: Study on antigenicity of the outer-membrane-protein of Bordetella aviumTo explore extraction and identification of outer membrane protein of Bordetella avium. We could determine contents of Bordetella avium OMP with Bradford, do SDS-PAGE electrophoresis of Bordetella avium OMP and prepare OMP immunity to immunity one-day-chickens with three different doses. The result showed that: the contents of Bordetella avium OMP was 300μg/mL; the best immunity- dose was 0.5ml/each; chickens that antibody titer was hyper-28 could fight back the lethal dose of Bordetella avium by testing between immunity-antibody and protection after invasion. We can infer that the lasting-time of antibody was enough to protect one- month-chickens.It can be inferred that the outer membrane protein of Bordetella avium had antigenicity and was worthy of preparation for its polyclone antiserum and diagnosis for infection of Bordetella avium and research of subunit vaccine of Bordetella avium.