新型冠状病毒S蛋白Del 143～145突变核酸标准物质的制备及定值研究
Development and quantification of nucleicacid reference material of SARS-CoV-2 S protein with Del 143-145 mutation
梁志超 1赵兴春 2王晓文 3徐雷 3孟昊洋 3鲁玺龙 2赵洁 4冯兆民 1李夫 1王全意 5张代涛 1杨扬仲夫 3潘阳6
作者信息
- 1. 北京市疾病预防控制中心传染病地方病控制所 100013
- 2. 公安部物证鉴定中心,北京 100038
- 3. 中国计量测试学会行政审批部,北京 100125
- 4. 北京睿信捷环保科技有限公司,北京 100010
- 5. 北京市疾病预防控制中心中心办公室 100013
- 6. 国家药品监督管理局药品审评中心生物制品临床部,北京 100176
- 折叠
摘要
目的 研制新型冠状病毒S蛋白Del143～145突变位点假病毒核糖核酸标准物质,满足公共卫生检测领域对标准物质的需求.方法 以新冠病毒原始株序列为参考,引入Omicron变异株特征性S蛋白Del 143～145,使用三质粒慢病毒包装系统制备假病毒.对产物进行均匀性、长期稳定性和短期稳定性评价,多家实验室联合定值.结果 制备的假病毒标准物质均匀性良好,可在-40℃下长期保存6个月以上,在常温条件下稳定12 h以上.多家实验室联合确定标准物质量值为(1.37±0.24)× 104 copies/μL.结论 研究制备的标准物质均匀稳定,适用于相关实验室的量值传递;质量检定、质量控制等多个领域.
Abstract
Objective To developed the pseudovirus carrying point mutation of Del143-145 in S protein of severe acute respiratory coronavirus 2(SARS-CoV-2)as a standard reference material(SRM)of nucleic acid,so as to meet the needs of SRM in public health practice.Methods The Del143-145 in S protein as the characterized mutation of omicron variant was imported into the genome of original SARS-CoV-2 which was used as the reference genome.The pseudovirus was then prepared based on a three plasmids lentiviral packaging system.The homogeneity,long-term and short-term stability were tested,and the concentration was determined by multiple laboratories.Results The SRM showed good homogeneity which was stable for more than 6 months at-40℃ and for more than 12 hours at room temperature.Based on multiple laboratories calculation,the value of SRM was 1.37±0.24×104 copies/μL.Conclusions The SRM prepared in the study had good homogeneity and stability,and is suitable for multiple fields,including dissemination of values of quantity,verificatin of instruments and quality control.
关键词
新型冠状病毒/慢病毒/核酸检测/标准物质Key words
Severe acute respiratory coronavirus 2/Lentivirus/Nucleic acid test/Standard reference Material引用本文复制引用
基金项目
国家重点研发计划(2023YFC3041500)
国家重点研发计划(2023YFC0872400)
国家重点研发计划(2023YFC 3041400-5)
北京市科技计划(Z231100004823002)
出版年
2024
NETL
NSTL
万方数据