SARS-CoV-2基因组RNA标准物质的研制

doi:10.3969/j.issn.1000-1158.2021.02.20

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摘要以灭活的人源新冠病毒基因组RNA为候选材料,采用数字PCR方法对新冠病毒E基因、ORF1ab基因和N基因拷贝数浓度进行确定,研制了高、低两种浓度的SARS-CoV-2基因组RNA标准物质(GBW(E)091098和GBW(E)091099)。高、低浓度标准物质E、ORF1ab和N基因的拷贝数浓度标准值及扩展不确定度(k=2)分别为:(1.06±0.11)×10³copy/μL,(8.96±0.61)×10²copy/μL,(1.73±0.13)×10³copy/μL;(1.29±0.26)×10²copy/μL,(9.5±1.8)×10¹copy/μL,(2.05±0.31)×10²copy/μL。该标准物质涵盖了新冠病毒所有特征基因的全部序列,目前已在新型冠状病毒核酸检测方法开发、检测试剂盒以及检测实验室的质量控制等方面得到了广泛的应用。

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	王迪
	王志栋
	吴枭
	牛春艳
	董莲华
	戴新华
	高运华

关键词 ：计量学, 新型冠状病毒, 标准物质, 实时定量PCR, 数字PCR

Abstract：High and low concentrations of reference material of SARS-CoV-2 genomic RNA were developed(GBW(E)091098 and GBW(E)091099)based on the inactivated RNA extraction of the novel coronavirus from infected patients as the potential material,of which the concentrations of E,ORF1ab and N gene were qualified by digital PCR. The reference values with expanded uncertainty of high concentration of reference material were(1.06±0.11)×10³copy/μL,(8.96±0.61)×10²copy/μL and(1.73±0.1)×10³copy/μL,respectively. And those for the low concentration of reference material were(1.29±0.26)×10²copy/μL,(9.5±1.8)×10¹copy/μL and(2.05±0.31)×10²copy/μL,respectively. The reference material here developed had been widely applied to method establishment of nucleic acid detection for SARS-CoV-2,quality control for detection kits and testing laboratory.

Key words： metrology SARS-CoV-2 reference material real-time quantitative PCR digital PCR

收稿日期: 2020-07-27 发布日期: 2021-02-18

PACS:

TB99

基金资助:国家质量研发重点专项(2017YFF0204604);中国计量科学研究院基本科研业务费项目(31-AKY1950)

通讯作者: 高运华(1972-),山东菏泽人,中国计量科学研究院副研究员,主要从事生物计量与标准方面的研究。 Email: gaoyh@nim.ac.cn E-mail: gaoyh@nim.ac.cn

作者简介: 王迪(1986-),男,河北秦皇岛人,中国计量科学研究院博士后,主要从事生物计量方面的研究。Email: wangdi@nim.ac.cn

引用本文:

王迪,王志栋,吴枭,牛春艳,董莲华,戴新华,高运华. SARS-CoV-2基因组RNA标准物质的研制[J]. 计量学报, 2021, 42(2): 259-264.
WANG Di,WANG Zhi-dong,WU Xiao,NIU Chun-yan,DONG Lian-hua,DAI Xin-hua,GAO Yun-hua. Development of Reference Material of SARS-CoV-2 Genomic RNA. Acta Metrologica Sinica, 2021, 42(2): 259-264.

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