Establishment and application of multiplex PCR assays for BVDV,IBRV and AKAV
Bovine viral diarrhea virus(BVDV),infectious bovine rhinotracheitis virus(IBRV)and Akabane virus(AKAV)are the main pathogens that cause reproductive disorders in cattle.In this study,a method was developed for the detection of IBRV and AKAV which are the main patho-gens causing reproductive disorders in cattle.Based on the sequences of the 5'-UTR of BVDV,the gB gene of IBRV,and the S gene of AKAV,three specific primer pairs were designed and use to develop the multiplex PCR assay for BVDV,IBRV and AKAV.The results showed that the devel-oped multiplex PCR method did not amplify other viruses of bovine origin,and the minimum de-tection limits for BVDV,IBRV and AKAV were 103,103 and 104 copies/μL,respectively.184 clini-cal serum samples were examined for BVDV,IBRV,and AKAV.The positive rates of BVDV,IBRV and AKAV were 18.48％,14.13％and 1.63％,respectively.The positive rates of mixed infec-tions of BVDV and IBRV were 3.8％and 1.63％for BVDV and AKAV.These results were con-sistent with the entry-exit quarantine industry standard in China with a rate of more than 92％,and the rate of mixed infection of BVDV+IBRV and BVDV+AKAV was 100％.The establish-ment of the multiplex PCR assay for BVDV,IBRV and AKAV provides a rapid and simple molec-ular biological method for the detection and epidemiological investigation on BVD,IBR and AKA in cattle.
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