The Establishment And Preliminary Application Of PCR For Bovine Respiratory Disease Complex

(Bovine respiratory syncytial virus,BRSV),(Infections bovine rhinotracheitis virus,IBRV),(Bovine parainfluenza virus3,BPIV-3) are me main primary disease of respiratory tract pathogens.They can cause bovine respiratory disease complex BRDC,and they always cause mixed infection,BRDC is a very common disease,And it has a high rate of infection and a high rate of mortality.BRDC is one of the major problems at present,It caused serious economic losses to the cattle industry,the study on BRD has been a long-term global priorities. The cattle farmers association of the United States and other institutions think it that bovine respiratory disease complex is the most important disease in the world.Bovine respiratory disease complex is known as one of the three diseases in Europe.The infection rate and the incidence rate of bovine respiratory disease complex is rising in our country.Bovine respiratory disease complex caused serious economic losses of cattle industry.According to the BRSV N gene sequence, IBRV gB gene sequence and BPIV-3N gene sequence in GenBank,Three pairs of primers were designed with Primer5.0.Based on the single PCR,The reaction system was optimized,A multiplex PCR method was established for BRSV, IBRV and BPIV-3.At the same time this method amplified BRSV596bp,IBRV398bp and BPIV-3704bp size,But it tested negative for BAdV-7,Anaplasma marginale and Vero cells.The sensitivity of the PCR assay was BRSV1TCID50·100μL-1,IBRV10-2TCID50·100μL-1and BPIV-310TCID50·100μL-1The multiplex PCR method,duplex PCR method,BRSV single method PCR method,IBRV single method PCR method,,BPIV-3single method PCR method,isolation and identification methods for virus.They were used to test7clinical samples.The test is successful,The multiplex PCR method,duplex PCR method,BRSV single PCR method,IBRV single PCR method,,BPIV-3single PCR method,their results were identical,The isolation and identification methods for virus had a lower detection rate of positive results.lt suggests that the detection of PCR method for clinical application is valuable.They could be used to detect the three kinds of virus in the same time and differential.