Brucella LPS Extraction And Purification And Polyclonal Antibody Preparation

Brucellosis is caused by Brucella. It is also known as Mediterranean remittent fever andheat waves. As brucellosis is a zoonotic disease, and there is no good cure, brucellosiseconomic benefits of aquaculture animals caused some impact on human health and cause acertain degree of threat. Prevention and control of brucellosis epidemic is imminent.Countries are now actively studying effective means of prevention and treatment ofbrucellosis. So the various functional components of brucellosis research have becomeincreasingly important.Studies have shown that the immunogenic outer membrane protein andlipopolysaccharide(LPS) in the surface of Brucella, not only play a good immune effect, butalso greatly improve the safety of immunization. LPS can also be used in the detection ofbrucellosis. Therefore, this experiment extracted the LPS of Brucella abortus and LPS used asa surface antigen. The specific polyclonal antibody is prepared to detect and researchbrucellosis.1. In this study, use the optimized phenol-water-PCP extracted LPS swine brucellosisvaccine strain S2. Then the LPS was extracted and purified by the method ofenzymatic and over speed centrifugal.2. For the extracted and purified LPS content was measured, the protein content wasdetermined, LAL activity was detected.3. Using the extracted LPS and inactivated Brucella bacteria as antigen, then theBALB/c mice were immunized according to the procedure of immunization.Detection the antibody titers of serum in mice by indirect ELISA Methods.In this study, extraction and purification of LPS with swine brucellosis vaccine strain S2,then the BALB/c mice were immunized with LPS according to the procedure ofimmunization. Then we assay the mice serum with indirect ELISA, obtained the mouseserum which the titer of antibody reached1∶6400. This study had laid a certain foundationfor the clinical testing and diagnostic.