Rapid Diagnosis Of Infectious Bovine Rhinotracheitis

testing. This test is complicate in operation, and it needs long time and high costs,so it is not fit for grass-roots veterinarians.For rapid diagnosing of IBR,we would develop a polymerase chain reaction(PCR)assay to detect Infectious Bovine Rhinotracheitis Virus (IBRV) and develop a indirect enzyme-linked immunosorbent assay (ELISA) kit to detect antibodies against IBRV in serum.Based on the nucleotide sequence of the IBRV, specific primers were designed. The react conditions were optimized. IBRV genome fragments(362bp) can be amplified by the PCR,and it has no positive reaction with the system to amplify Pseudorabies virus(PRV), Herpesvirus of turkeys(HVT)and Bovine viral diarrhea virus(BVDV). Sensitivity assay indicate the system can detect the 711TCID50.The indirect ELISA for detection antibodies of IBRV was established. Antigen as coated was strain Bartha Nu/67 virus collected after inoculating into primary goat kidney cell. This assay optimal working dilution was ascertained, including antigen, testing sera, mouse anti-bovine monoclonal antibody, rabbit anti-mouse immunoglobulin conjugated to horseradish peroxidase.The standard of determining as positive sample was established. Using this method detected 138 sera samples, results indicate the specificity of the test is 93.5%,the sensitivity is 93.5% ,the agreement is 93.5% ,compared with the virus neutralization testing.The agreement is 97.6%, compared with the IBR ELISA kit from IDEXX.Using this method to detected 230 bovine sera samples,168 samples was positive,the agreement is 87.8%, compared with the IBR ELISA kit from IDEXX。Using this method to detected 35 yak sera samples infected by IBRV from Tibet, 21 samples was positive, the agreement is 85.7%, compared with the virus neutralization testing.