Establishment And Primary Application Of PCR Method To Diagnose IBR

The study succeeds in designing and synthesizing a pair of primers according to thepublished sequence of the Infectious Bovine Rhinotracheitis Virus (IBRV). The primers areP,-5'-GAGGAAGAGGAGGAGTTTGACG-3',P2-5'-AGCCGCAAATAACACGGTGTGC-3'.It has shown that the PCR systemic has premium conditions , that is , thedenaturing temperature is 94 ℃ , the anneal temperature is 56 ℃ ,the circles are 35times .respectively. The premium results of the PCR system are that the primerconcentration is 25 pmol ,dNTP dosage is 2ul , Taq polymerase dosage is 1 unit , MgCbconcentration is 3.0mM.The PCR can amplify 450bp specific IBRV fragments. It accords with the predictedamplification segament. Howerver , it has no positive result with the system to amplifyBovine viral diarrhea virus,Bovine Rotavirus, Bovine coronavirus. So ,it can demonstratethe PCR amplification system has good specificity. In addition , it also possesses very highsensitivity because the system can detect the 10~6dilute template.The primary application with the PCR system in clinic has shown that the positive rate is 30c/o, which is basically identical with other virus isolation methods. According to theabove results, the study has developed an IBRV PCR detection method, which possesseslatent application prospect.