Isolation and Identification of Bovine viral diarrhea virus Strain GSLY

doi:10.7668/hbnxb.20190722

Abstract

Abstract: To understand the prevalence and molecular characteristic of Bovine viral diarrhea virus (BVDV)in Gansu Province,stool samples of cattle with diarrhea symptoms in a cattle farm in Gansu Province were collected and detected for BVDV by RT-PCR.The stool samples identified as BVDV positive were treated and then inoculated into MDBK cells for virus isolation and culture.Subsequently,the isolated virus was identified by RT-PCR,immunofluorescence detection and viral gene sequencing,and genetic evolution analysis was then carried out.The result revealed that a BVDV strain was successfully isolated and named BVDV-GSLY.BVDV-GSLY was identified as a cytopathogenic(CP)biotype since BVDV-GSLY could produce cytopathic effect(CPE)on MDBK cells.The 5'-UTR and the N^pro of BVDV-GSLY were amplified by RT-PCR and the PCR products were the expected sizes,respectively.Immunofluorescence assay showed that BVDV-GSLY could react with BVDV antibody to generate specific fluorescence.The virus titer was 10^7.8TCID₅₀/0.1 mL when BVDV-GSLY passaged 12 times in MDBK cells.The genetic evolution analysis of the 5'-UTR and N^pro sequences indicated that BVDV-GSLY was closely related to SD1 strain,and the homology with SD1 strain was 95.5% in 5'-UTR and 91.5% in N^pro,respectively.The phylogenetic tree also showed that BVDV-GSLY was in the same branch with SD1 strain,indicating that BVDV-GSLY belonged to BVDB-1a subgenotype.A BVDV subgenotype 1a strain was successfully isolated,and the research enriched the molecular epidemiology of BVDV and provided the theoretical foundation for prevention and control of BVDV in Gansu Province and the development of BVDV vaccine.

Key words: Bovine viral diarrhea virus(BVDV), BVDV-GSYL strain, Isolation, Identification, Phylogenetic analysis

CLC Number:

ZHANG Kang, ZHANG Kai, WANG Lei, CUI Dongan, ZHANG Jingyan, YAN Zunxiang, MA Xueqing, XUE Huan, LI Jianxi. Isolation and Identification of Bovine viral diarrhea virus Strain GSLY[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(S1): 378-384. doi: 10.7668/hbnxb.20190722.

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