一株源于商品胎牛血清的BVDV-2型的分离鉴定及基因组序列分析

doi:10.11843/j.issn.0366-6964.2020.02.013

Abstract

Abstract: Bovine viral diarrhea virus (BVDV) is an important pathogen causing bovine viral diarrhea/mucosal disease, and it is also the common source of contaminant in bovine serum and related biological products. In this study, a cytopathogenic (CP) type BVDV (GS2018 strain) was isolated from commercial FBS and identified by transmission electron microscopy (TEM), molecular identification, immunofluorescence assay, complete genome sequencing and genetic evolution analysis respectively. The results showed that MDBK cells presented obvious cytopathic effect (CPE) when inoculated with the BVDV GS2018 strain, and the virus titer was up to 10^6.2·0.1 mL^-1. The purified virus was detected by transmission electron microscope (TEM), round-shaped virions were observed with the diameter ranging from 50 to 60 nm, and specific positive fluorescence signal was found clearly by immunofluorescence assay. Meanwhile, the complete genome of GS2018 was sequenced as 12 235 nucleotides (nt), and BLAST result of 5'UTR region revealed that the strain had high homology with the reference sequences of BVDV-2. The phylogenetic evolutionary analysis of 5'UTR, N^pro and E2 genes showed that the GS2018 strain was classified into BVDV-2a subtype obviously, sharing the highest homology (up to 98%) with BVDV-2a representative strain USMARC-60764. Moreover, an interesting phenomenon was found that no nucleic acid fragment insertion was present in the NS2/3 region with further analysis of the genome, unlike most of CP type BVDV-2. The diversity of BVDV2 genome suggested that the mechanism of BVDV induced cytopathic effect is complicated and further research of genomic variation will contribute to answering this question.

Key words: BVDV-2, cytopathic effect, complete genome, phylogenetic evolution analysis

CLC Number:

S852.659.6

LI Yanping, WANG Liqun, LI Huixia, LIANG Panhong, CAI Xuepeng, MAO Li, LUO Xuenong. Identification and Complete Genome Sequencing Analysis of a Cytopathic BVDV-2 from Commercial Fetal Bovine Serum[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(2): 320-328.

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Identification and Complete Genome Sequencing Analysis of a Cytopathic BVDV-2 from Commercial Fetal Bovine Serum

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