水貂阿留申病毒核酸标准物质的研制
Preparation of nucleic acid reference materials of Aleutian mink disease virus
王春霞 1商金源 1吴顺 1闫曼平 1冯二凯 1罗国良 1易立 1叶京飞 1赵宗正 2高华 3程悦宁 1王振军1
作者信息
- 1. 中国农业科学院特产研究所,吉林长春 130122
- 2. 军事科学院军事医学研究院军事兽医研究所,吉林长春 130122
- 3. 通辽市科尔沁区动物疫病预防控制中心,内蒙古通辽 028000
- 折叠
摘要
为研制均匀、稳定的水貂阿留申病毒(AMDV)核酸标准物质,选择水貂阿留申病毒AMDV-G株接种CRFK细胞进行病毒繁殖,经灭活后,制备水貂阿留申病毒核酸检测标准物质.利用微滴式数字PCR方法对制备的核酸标准物质进行均匀性检验、稳定性评估及联合8家实验室进行合作定值.均匀性检验结果:F=1.56＜Fa(F(0.05,14,30)=2.04),表明本标准物质组内和组间无统计学差异.稳定性评估结果表明:本标准物质在-20 ℃环境中可稳定保存6个月、在4 ℃环境中可稳定保存9 d、在-20 ℃环境下可稳定冻融4次.经8家有实验室检测资质认证的实验室合作定值及不确定度评定,最后得到本标准物质的定值为1.05× 104 copies/μL,扩展不确定度为0.14× 104 copies/μL.本标准物质定值准确、均匀性好、量值稳定,可用于水貂阿留申病毒核酸检测等相关实验,有利于水貂阿留申病的诊断和防控.
Abstract
In order to prepare a uniform and stable nucleic acid reference material for Aleutian mink disease virus(AMDV),AMDV-G strain was selected and inoculated into CRFK cells for virus reproduc-tion,after inactivation,the nucleic acid reference material was successfully prepared.We used the droplet digital PCR method to conduct homogeneity and stability,and collaborative calibration with 8 laboratories on the prepared nucleic acid reference materials.The homogeneity test showed that F=1.56＜Fa(F(0.05,14,30)=2.04),indicated that there was no statistical difference within or between groups of this nucleic acid reference materials.The stability evaluation results indicated that,this nucleic acid reference materials can be stable for 6 months at-20 ℃,9 days at 4 ℃,and 4 freeze-thaw cycles at-20℃.Collaborated with 8 laboratories with laboratory testing qualifications for fixed value and uncertainty assessment,the quantity value was 1.05 × 104copies/μL,the expanded uncertainty was 0.14 × 104 copies/μL.In summary,the prepared nucleic acid reference materials were advantaged by accurate calibration,great homogeneity and stability evaluation,it can be used for related experi-ments such as AMDV nucleic acid detection,which is beneficial for the diagnosis,prevention and control of Aleutian mink disease.
关键词
水貂阿留申病毒/核酸标准物质/微滴式数字PCRKey words
Aleutian mink disease virus/nucleic acid reference material/droplet digital PCR引用本文复制引用
基金项目
中国农业科学院科技创新工程项目(CAAS-ASTIP-2021-ISAPS)
吉林省科技创新平台项目(YDZJ202302CXJD034)
吉林省科技发展计划项目(20210202099NC)
吉林省发改委项目(2022C042-9)
出版年
2024
国家科技期刊平台
NSTL
维普
万方数据