The Direct Rapid Immunohistochemistry Test (DRIT) For The Detection Of Rabies Virus Antigen

Rabies is an amphixenosis caused by rabies virus.People and all warm-blooded animals are susceptible to rabies.Because once the disease is incurable disease,and because the widespread existence of animal infection,control is very difficult,so the diagnosis of human and animal rabies infection is very important.The Direct Rapid Immunohistochemistry Test(DRIT)is a newly developed method for the detection of rabies virus antigen using enzyme-labeled anti-rabies virus monoclonal antibody in US CDC rabies room.According to the national standard operating procedures for the diagnosis of rabies virus animals,DRIT is a standard for validated d FA methods that have not yet been certified.US CDC rabies room evaluation experiments show that DIRT and WHO recommended "gold standard" DFA test results in line with the rate of 100%.This reagent is our company and the Chinese Center for Disease Control and Prevention(CDC)of viral disease prevention and control of the results of cooperative research.Anti-rabies virus monoclonal antibody cell lines,rabies virus positive brain tissue slide and negative brain tissue slide are provided by CDC.The unit of monoclonal antibody cells were cultured,monoclonal antibody purification and labeling,the antibodies were detected by immunohistochemical method in rabies virus antigen,the experimental method was optimized,select the best experimental program,supporting experimental materials,consumables and Packaging,all materials assembled into kits.Operation process is as follows:Thawing The Rabies virus antibodies cell line(No: 4A12)and culture it.Insert cells into the Balb/c mice abdominal cavity to get ascites.The recycling of ascites was using indirect ELISA method to detect the antibody titer has the high titer detected.Salting out purified antibody and then labeled avidin-biotin system of monoclonal antibody.Using indirect ELISA method for detecting labeled antibody titer has the high titer detected.Using the cell plate vaccination that has rabies virus to test,and determines the feasibility of this experimental method in detecting rabies virus disease.Screening best work concentration of the antibody biotin markers and SA-HRP.The best work concentration of the antibody biotin marker is 1:100.And the best work concentration of SA-HRP is 1:1000.And then detect rabies virus positive brain tissue prints and negative brain tissue prints,the method can identify the positive and negative.Finally,the all of the reagents required for this test,materials and consumables assembled into kits after partial shipments.In this experiment the Direct Rapid Immunohistochemistry Test(DRIT)established this monoclonal antibody can specificity combined with rabies virus and negri bodies,and with optical microscope can be observed within the brain tissue of negri bodies.So,the reagent can be applied to Direct Rapid Immunohistochemistry Test(DRIT)to detect rabies virus antigen,and greatly simplified with experiment,saves the experiment cost,improve the specificity and repeatability of the experiment.At present,domestic also did not use the method of detection of rabies virus finished product.We can study of the reagents for further,and make the Direct Rapid Immunohistochemistry Test(DRIT)for the detection of rabies virus antigen is widely used in the laboratory.