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目的 为了掌握近期我国牛群间牛疱疹病毒4型(Bovine herpesvirus 4,BoHV-4)的分子演化特征与生物学特性，并评估对非天然宿主中妊娠个体的感染致病性。方法 采集某奶牛场流产奶牛的阴道拭子及血液样品，采用PCR方法检测BoHV-4和其他牛常见易感病毒；将BoHV-4阳性样品接种MDBK细胞进行病毒分离鉴定，通过测定TCID50、绘制多步生长曲线、分析主要基因(gB、gH和TK)遗传演化分析，明确分离毒株的基本分子生物学特征；将病毒液(106.5TCID50/0.1 mL)以滴鼻+肌注的方式接种妊娠中期的母兔，滴鼻和肌注各0.5 mL/只，观察其感染与致病能力；对流产奶牛进行6个月随访，监测其BoHV-4抗体水平及病原携带情况，探究该病毒对母牛繁殖能力的持续影响。结果 仅流产奶牛的阴道拭子检测出了BoHV-4基因，成功分离获得1株BoHV-4流行株，命名为BoHV-4 GSHZ/06/2025株；该毒株第7代病毒接种细胞后约72 h达到复制高峰。遗传演化分析表明，该毒株基于TK基因可归属Genotype 1，基于gB和gH基因可归属Genotype 2，且重组分析提示其gH基因可能为Genotype 1和Genotype 2病毒重组演化产生。动物感染试验显示，滴鼻+肌注的联合接种方式未能使该分离毒株感染妊娠母兔。随访结果显示，20头流产奶牛6个月后阴道拭子和血清中BoHV-4核酸均为阴性，而血清BoHV-4抗体阳性率达100%。结论 本研究成功分离的BoHV-4流行毒株不同基因存在明显的遗传演化差异，其发生的重组事件极有可能改变病毒感染致病力；该毒株在妊娠母兔模型中未建立有效感染，提示BoHV-4更可能作为机会性或协同致病因子参与牛生殖系统疾病的发生。本研究从流产的奶牛上成功分离到1株新的BoHV-4流行株，提示我们需重新审视BoHV-4感染对母牛繁殖性能的影响，其潜在的危害性值得重点关注，本研究结果也为BoHV-4的流行防控及其相关研究提供了基础数据。
Abstract：Objective To characterize the molecular evolutionary features and biological properties of bovine herpesvirus type 4(BoHV-4) circulating in cattle herds in China, and to evaluate its infectivity and pathogenic potential in a non-natural pregnant host. Methods Vaginal swabs and blood samples were collected from aborted dairy cows on a commercial farm. BoHV-4 and other common bovine pathogens were screened by PCR. BoHV-4 – positive samples were inoculated onto Madin – Darby bovine kidney(MDBK) cells for virus isolation. Viral characteristics were assessed by determining the 50% tissue culture infectious dose(TCID50), constructing multi-step growth curves, and performing phylogenetic analyses based on the glycoprotein B(gB), glycoprotein H(gH), and thymidine kinase(TK) genes. To assess cross-species infectivity, a virus suspension(106.5 TCID50/0.1 mL) was administered to mid-gestation pregnant rabbits via combined intranasal and intramuscular inoculation(0.5 mL per route per animal). In addition, aborted cows were followed for six months to monitor BoHV-4 antibody levels and viral persistence, in order to evaluate the potential long-term effects on reproductive performance. Results BoHV-4 DNA was detected exclusively in vaginal swabs from aborted cows. One BoHV-4 field strain, designated BoHV-4 GSHZ/06/2025 strain, was successfully isolated. The seventh passage reached peak replication at approximately 72 h post-infection in MDBK cells. Phylogenetic analysis indicated that the isolate clustered within Genotype 1 based on the TK gene, but within Genotype 2 based on the gB and gH genes, suggesting a mosaic genomic structure. Recombination analysis further indicated that the gH gene may have arisen from recombination between Genotype 1 and Genotype 2 lineages. Experimental infection showed that combined intranasal and intramuscular inoculation did not establish productive infection in pregnant rabbits. Follow-up results demonstrated that all vaginal swabs and blood samples from 20 aborted cows were negative for BoHV-4 nucleic acid six months later, whereas the seropositivity rate for BoHV-4 antibodies reached 100%. Conclusion The BoHV-4 strain isolated in this study exhibits distinct genetic heterogeneity across genomic regions, likely resulting from recombination events that may influence viral biological properties. The failure to establish infection in a pregnant rabbit model suggests that BoHV-4 may act as an opportunistic or co-infecting pathogen in bovine reproductive disorders. These findings highlight the need to re-evaluate the role of BoHV-4 in cattle reproductive health and provide a foundation for future epidemiological surveillance and control strategies.
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.260039
中图分类号:S852.653
引用信息:
[1]赵妍,张财祯,王辉,等.一株牛疱疹病毒4型流行株的分离鉴定及其分子演化特征分析[J].病毒学报,2026,42(03):923-937.DOI:10.13242/j.cnki.bingduxuebao.260039.
基金信息:
甘肃省科技计划项目（项目号：25CXNA034）,题目：甘南耗牛重要病毒性传染病的流行病学调查及防控关键技术示范应用;甘肃省科技计划项目（项目号：25CXNA045）,题目：甘肃省牛呼吸道和消化道主要病毒性传染病的分子流行病学调查及防控技术培训; 中国农业科学院科技创新工程（项目号：CAAS-AS-TIP-2021-LVRI）,题目：宿主抗病毒感染与免疫生物学研究~~
2026-05-11
2026-05-11
2026-05-11