山东地区1株牛病毒性腹泻病毒的分离鉴定及遗传进化分析

doi:10.16431/j.cnki.1671-7236.2024.10.029

Abstract

Abstract: 【Objective】 This study was aimed to understand and master the biological characteristics and genetic variation of Bovine viral diarrhea virus(BVDV) epidemic strains in Shandong province，and provide theoretical basis and technical support for the prevention and control of BVDV.【Method】 The tissue supernatant was prepared from suspected BVDV-infected samples of a cattle farm.RT-PCR method was used for pathogen identification，MDBK cells were inoculated for virus isolation and culture，and then subjected to indirect immunofluorescence assay(IFA) and electron microscopy for identification，and the whole genome of the isolated strain was segmented and amplified，cloned and sequenced.The alignment and analysis of genetic evolution of the whole genome，N^pro and E2 genes were done by Lasergene and other softwares.【Result】 The RT-PCR results of BVDV-specific primers showed BVDV-specific bands of approximately 504 bp，which was consistent with the expected band size.The IFA results showed specific green fluorescence,and virus particles about 50 nm in diameter were visible by electron microscopy，indicating that a non-cytopathogenic BVDV strain named SDNF9 was isolated.The total sequence of the strain was 12 232 nt.Comparing to BVDV reference strains，the sequence similarity was 79.1% to 93.8%，with the highest homology of 93.8% observed with the 22AH-1 strain from China and the Bega-like strain from Australia.Multiple amino acid variations were found in the N^pro and E2 proteins.Phylogenetic analysis showed that SDNF9 was in a different branch of genetic evolution from the BVDV2 and BVDV3 strains，while it was in the same clade as BVDV1 and belonged to the BVDV 1c genotype.【Conclusion】 In this study，an epidemic strain of BVDV 1c was successfully isolated and genomic and genetic evolution analysis was carried out，it was closely related to Chinese isolated 22AH-1，which provided data and material support for the prevention，control and vaccine development in the region.

Key words: Bovine viral diarrhea virus (BVDV); isolation; identification; sequence analysis; phylogenetic analysis

CLC Number:

S852.65⁺3

BIJunming, DONG Yaqin, CUIJin, SHA Zhou, GU Congcong, ZHENG Hui, WEI Rong, SUN Fuliang, ZHANG Feng, ZHANG Zhihong. Isolation，Identification and Genetic Evolution Analysis of a Strain of Bovine Viral Diarrhea Virus in Shandong Province[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(10): 4485-4499.

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Isolation，Identification and Genetic Evolution Analysis of a Strain of Bovine Viral Diarrhea Virus in Shandong Province

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