IBRV GB Monoclonal Antibody Preparation And Blocking ELISA Detection Methods Established

Infectious bovine rhinotracheitis virus(IBRV), can violations of multiple tissues and organs. It displays the different clinical type, such as respiratory type, genital type, meningitis type,etc.When the virus invades the animal, they can Lurk in a certain area. When the resistance is low it can discharge poison outside. It is caused great threat of cattle allover the world.The disease was first discovered at the US state of Colorado in 1955, then there are some reports about the disease in many countries. The disease was introduced into China in the 20th century 80 years. At present, the domestic haven't standard test method, the diagnosis of IBR mainly rely on foreign expensive kit, so need to develop effective prevention and accurate diagnosis IBR new product Infectious.IBRV genome can coding of 11 glycoprotein, including gB protein is virus of membrane surface show cystic main protein, and also one of the main immune original protein in the virus to cause disease, stimulates the production process and the process of antibodies are a greater role, because gB protein can produce protective a.granoff trigger many researchers in the diagnosis of its made a lot of research methods.We can raise IBRV and purify by using the sucrose gradient centrifugation. Application kit to deteect protein content is 5.7083 mg/ml,5.7083mg/ml, TCID50 is 10-533/0.1ml. Using PCR method to test purify, The results prove that we acquire high purity of IBRV. Use the purified virus immune BALB/C mice three times, and IBRVgB strengthen the immune one time. Testing positive serum titer before each immunity. Eventually, they all reached more than 1:12800.This explains the mice immune effect is good, can be used for fusion. Use 50% of PEG4000 as fusion agent. Three experimental results more than 70% in six times fusion experiments.Fusion rate is more higher.Using whole virus and IBRVgB to screens, unselective we won a monoclonal antibody named 2C4, and Purify it by Affinity chromatograph. Protein content is 3.647 mg/ml. Ascites titer is 1:12 800.It can proof that the monoclonal antibody has high specificity by SDS-PAGE electrophoresis, Western-bolt analysis specific test.This can establish a blocking ELISA test method. Working conditions:antigen3.603μg/ml, monoclonal antibody 9.116μg/ml. Do cross test about soms other kinds of Positive serum of cow's common diseases and infected positive serum artificially infected. The results are same tol indirect ELISA. We can draw the conclusion:this method has good specificity and sensitivity, suitable for the diagnosis and IBR epidemiology investigation for the disease, monitor provides a kind of effective method.