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作 者:邓雅丽 谢玲玲 付杰 周敏 张艳芳[1] 胡思婧 邓菲[1] DENG Yali;XIE Lingling;FU Jie;ZHOU Min;ZHANG Yanfang;HU Sijing;DENG Fei(Wuhan Institute of Virology,Chinese Academy of Sciences,Wuhan 430207,China)
机构地区:[1]中国科学院武汉病毒研究所,湖北武汉430207
出 处:《生物化工》2024年第3期94-97,共4页Biological Chemical Engineering
基 金:国家科技资源共享服务平台项目(E1YZ020501);湖北省农业种质资源保护与利用项目(HBZY2023A001-16)。
摘 要:目的:通过对5例疑似感染鼠痘病毒的实验小鼠脑组织进行病毒分离培养、分子生物学鉴定和形态学研究,进一步深入探索鼠痘病毒的生长特性与形态学特征。方法:从感染的鼠脑组织中分离鼠痘病毒,针对鼠痘病毒基因设计特异性引物,建立一种实时荧光定量PCR检测方法,对病毒进行检测鉴定;利用非洲绿猴胚胎肾细胞(Marc145细胞),通过电镜切片观察和生长曲线测定,考察病毒的形态特征和感染特征。结果:成功分离得到5株鼠痘病毒,病毒感染Marc145细胞后发生相应病变效应,在感染后0~72 h病毒滴度逐渐升高并在72 h达到最高值,随后滴度逐渐降低,但在感染96 h仍然保持较高病毒滴度;5株鼠痘病毒毒株在整体的生长特性与形态学方面具有相似性。结论:研究结果为深入探究鼠痘病毒的生命周期、感染机制、病毒结构和宿主免疫反应等病毒学基础提供了基础。Objective:To further explore the growth and morphological characteristics of ectromelia virus by isolating and culture,identifying molecular biology and morphological aspects of the brain tissues of five experimental mice suspected of being infected with ectromelia virus.Methods:The ectromelia virus is isolated from the infected mouse brain tissue,and specific primers are designed for the ectromelia virus gene,and a real-time fluorescence quantitative PCR detection method is established to detect and identify the virus.African green monkey embryonic kidney cells(Marc145 cells)are used to observe the morphological characteristics and infection characteristics of the virus by electron microscopy section observation and growth curve determination.Results:Five strains of ectromelia virus are successfully isolated,and the corresponding lesion effect occurres after virus infection with Marc145 cells,and the viral titer gradually increases at 0 h to 72 h after infection and reaches a peak at 72 h,and then the titer gradually decreases,but still maintains a high viral titer at 96 h after infection.The five strains of ectromelia virus are similar in terms of overall growth characteristics and morphology.Conclusion:The results of this study provide a basis for in-depth exploration of the virologic basis of ectromelia virus,such as life cycle,infection mechanism,viral structure and host immune response.
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