1株牛结节性皮肤病病毒的分离鉴定与生物学特性研究

doi:10.16431/j.cnki.1671-7236.2025.06.031

Abstract

Abstract: 【Objective】 The objective of this study was to investigate the etiological characteristics of bovine Lumpy skin disease virus (LSDV) in Shandong.【Method】 Skin nodules of suspected lumpy skin disease (LSD) cattle were collected from a cattle farm in Shandong province,and primary lamb testicular cells (PLT) were used for virus isolation.The LSDV G-protein-coupled chemokine receptor (GPCR) gene was amplified by PCR and sequenced,and the virus specificity was identified by indirect immunofluorescence assay (IFA) using anti-LSDV specific serum.The genome of the isolated strain was sequenced and its genetic variation was analyzed.The isolated strain was passed continuously in different cells,the virus titers were measured,and the growth curve was drawn.The proliferation ability in different cells was further determined.The neutralization index of the isolated strain against the antibodies of goat pox vaccine was determined by antibody neutralization test.【Result】 The nodular tissue samples of diseased cattle were cultured by PLT cells,and after blind transmission for 3 generations,the cytopathic effect (CPE) characterized by cell roundness,aggregation and shedding occurred.PCR amplification obtained 1 010 bp specific band of GPCR gene of the isolated strain，and the similarity of this gene sequence with LSDV strains at home and abroad was 99.3%-100%.The isolated strain could bind to the specific serum against LSDV,and the specific green fluorescent spot appears after immunofluorescence staining.The results of genome sequencing showed that the isolated strain was genetically close to the 7 domestic strains in 2019-2022 and Thailand strain in 2022.The virus titer was stable at 10^-6.08 to 10^-6.17 TCID₅₀/mL when the isolates were passed through 20-30 generations.The growth curve showed that the titer of the isolated strain peaked at 96 h (10^-6.16 TCID₅₀/mL) and decreased to 10^-5.60 TCID₅₀/mL at 144 h.The isolated strain could prolifate continuously on MBDK and BHK-21 cell lines for 3 generations,and the virus titers could reach 10^-4.85 and 10^-4.63 TCID₅₀/mL,respectively.The neutralization index of goat pox vaccine antibody against this strain (276) was significantly lower than that of goat pox vaccine strain (1 833) (P＜ 0.05).【Conclusion】 A strain of LSDV was successfully isolated from the skin nodular tissue of infected cattle.This strain was genetically close to other strains in China except Tibet,and could be stably cultured in MBDK and BHK-21 cells.Moreover,the neutralization ability of goat pox vaccine antibody against this strain was poor.

Key words: Lumpy skin disease virus (LSDV); isolation and identification; biological characteristics; vaccine

CLC Number:

S858.23

ZHAO Yilong, TANG Na, JING Changhua, XU Qingqing, YIN Xiusheng, WANG Haiming, SUN Jing, LIN Panpan, DONG Lin, LIU Jishan, CAO Rongfeng. Isolation,Identification and Biological Characteristics of a Strain of Lumpy Skin Disease Virus[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(6): 2790-2799.

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Isolation,Identification and Biological Characteristics of a Strain of Lumpy Skin Disease Virus

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