狂犬病病毒LN34基因假病毒核酸国家标准物质的研制  

Development of the National Nucleic Acid Reference Material of Rabies Virus LN34 Gene

作　　者：鞠厚斌[1] 李鑫[1] 葛菲菲[1] 杨德全[1] 沈海潇 王建[1] 赵洪进[1] Ju Houbin;Li Xin;Ge Feifei;Yang Dequan;Shen Haixiao;Wang Jian;Zhao Hongjin(Shanghai Center for Animal Disease Prevention and Control,Shanghai 201103,China)

机构地区：[1]上海市动物疫病预防控制中心,上海201103

出　　处：《中国动物检疫》2023年第4期123-129,共7页China Animal Health Inspection

基　　金：上海市科技兴农项目(2020-02-08-00-03-F01460);上海市“科技创新行动计划”技术标准项目(20DZ2200200)。

摘　　要：LN34基因作为狂犬病病毒核酸诊断的首选基因已被广泛应用,但检测活动中涉及生物安全要求以及缺少相应的核酸标准物质,导致很难对现有核酸诊断方法进行有效评价。本研究以慢病毒为表达载体,制备了含有狂犬病病毒LN34基因的假病毒核酸标准物质,并对该标准物质开展了均匀性、稳定性评估以及数字PCR法定值、不确定度分析和临床试用。结果显示:制备的标准物质定值为(1.33±0.23)×10^(4)拷贝/μL,样品均匀;-20℃条件下可稳定保存6个月,4℃可稳定保存9 d,25℃可稳定保存1 d。本标准物质的研制为相关实验室开展量值溯源、能力验证、质量控制、方法评价、试剂选择等工作提供了技术支撑。LN34 gene has been widely used as a priority to diagnose rabies virus,but it was difficult to effectively evaluate current methods for nucleic acids because of biosafety requirements and lack of corresponding nucleic acid reference materials during detection.In the paper,taking the lentivirus as an expression vector,a kind of nucleic acid reference material containing rabies virus LN34 gene was prepared,followed by evaluation on its homogeneity and stability,value determination by digital PCR assay,uncertainty analysis and clinical trial.It was found that,for the prepared material,its value was determined as(1.33±0.23)×10^(4) copies/μL,and samples collected therefrom were uniform;it could be stored stably for 6 months at-20℃,9 days at 4℃,and 1 day at 25℃.Relevant laboratories were technically supported by the development of the material to conduct traceability of quantity value,proficiency testing,quality control,method evaluation,reagent selection and other tasks.

关 键 词：狂犬病 核酸扩增 LN34基因 标准物质 

分 类 号：S852.6[农业科学—基础兽医学]