新型冠状病毒核酸检测方法及标准物质的研究进展

赵雨佳; 廖湉毅; 范培蕾; 梁亮; 赵海波; 沈正生

doi:10.12338/j.issn.2096-9015.2020.0375

新型冠状病毒核酸检测方法及标准物质的研究进展

doi: 10.12338/j.issn.2096-9015.2020.0375

赵雨佳^{1, 2,},
廖湉毅³,
范培蕾¹,
梁亮¹,
赵海波¹,
沈正生^1, ,

1.
北京市计量检测科学研究院，北京 100029
2.
清华大学药学院，北京 100084
3.
巴黎大学科学学院，巴黎 75006

基金项目: 国家重点研发计划（2019YFF0216703）；第66批中国博士后科学基金(2019M660539)。

详细信息

作者简介:
赵雨佳（1989-），北京市计量检测科学研究院博士后，研究方向：核酸计量、病原微生物检测方法等，邮箱：zhaoyj@bjjl.cn

通讯作者:
沈正生（1961-），北京市计量检测科学研究院教授级高工，研究方向：化学计量，邮箱：shenzs@bjjl.cn

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出版历程
- 网络出版日期: 2021-11-19
- 刊出日期: 2022-01-24

Perspective on SARS-CoV-2 Detection and Reference Materials

1.
Beijing Institute of Metrology, Beijing 10029, China
2.
Pharmacy College, Tsinghua University, Beijing 100084, China
3.
Science College, Paris University, Paris 75006, France

摘要

摘要: 新型冠状病毒（SARS-CoV-2）具有很强的传染能力，已造成全球大流行，对人类健康和社会的经济发展造成极大影响。快速、准确的检测新型冠状病毒对病情的确诊和疫情的防控尤为重要。我国国家市场监督管理总局已注册22种新型冠状病毒核酸检测试剂盒，能满足对高危人群的大规模筛查，有力保障疫情防控。同时，中国计量科学研究院研制的新型冠状病毒标准物质作为计量实体，进一步保障检测结果的准确性和溯源性。通过对国家食品药品监督管理局和国家标准物质资源共享平台公布的数据进行分析和梳理，结合国内外相关研究进展，就新型冠状病毒的检测方法、检测试剂盒及标准物质进行综述，以期为新型冠状病毒及其他传染性疾病的诊断和防控提供借鉴和参考思路。
- 计量学 /
- 新型冠状病毒 /
- 体外诊断 /
- 核酸检测 /
- 标准物质
Abstract: The novel coronavirus (SARS-CoV-2) has a strong ability to infect, which has caused the pandemic. It has seriously affected human health and life, social economy and development. Fast and accurate detection of the novel coronavirus is particularly important for the diagnosis, prevention, and control of the epidemic. Chinese CFDA has registered 22 nucleic acid detection kits, which can meet the needs of high-risk groups for large-scale screening and effectively guarantee the prevention and control of the epidemic. Meanwhile, the novel coronavirus reference material, as a measurement reference, developed by the National Institute of Metrology, China further guarantees the accuracy and traceability of test results. This article analyzes the data published by the State Food and Drug Administration and the National Standard Material Sharing Platform, and combines relevant domestic and international research results to systematically review the novel coronavirus detection methods, test kits and reference materials, hoping to shed light on diagnosis, prevention and control of the novel coronavirus and other infectious diseases.
- metrology /
- novel coronavirus /
- in vitro diagnosis /
- nucleic acid detection /
- reference materials

HTML全文

图 1 新型冠状病毒的核酸检测流程

Figure 1. The workflow of SARS-CoV-2 detection

下载: 全尺寸图片幻灯片

图 2 LAMP扩增示意图^[22]

Figure 2. The schematic diagram of LAMP^[22]

下载: 全尺寸图片幻灯片

表 1 CFDA注册的新型冠状病毒核酸检测试剂盒

Table 1. The registration SARS-CoV-2 detection kit of CFDA

序号	名称	注册证编号	适用范围
1	新型冠状病毒2019-nCoV核酸分析软件	20203210062	核酸检测分析软件
2	新型冠状病毒2019-nCoV核酸检测试剂盒（荧光PCR法）	20203400520	ORF1ab和N基因
3	新型冠状病毒（2019-nCoV）核酸检测试剂盒（荧光PCR法）	20203400299	ORF1ab和N基因、E基因
4	新型冠状病毒2019-nCoV核酸检测试剂盒（双扩增法）	20203400302	ORF1ab和E基因
5	新型冠状病毒2019-nCoV核酸检测试剂盒（荧光PCR法）	20203400322	ORF1ab基因、N基因
6	新型冠状病毒2019-nCoV核酸检测试剂盒（荧光PCR法）	20203400384	ORF1ab和N基因
7	新型冠状病毒2019-nCoV核酸检测试剂盒（荧光PCR法）	20203400065	ORF1ab和N基因
8	新型冠状病毒2019-nCoV核酸检测试剂盒（荧光PCR法）	20203400057	ORF1ab和N基因、E基因
9	新型冠状病毒2019-nCoV核酸检测试剂盒（荧光PCR法）	20203400060	ORF1ab基因
10	新型冠状病毒2019-nCoV核酸检测试剂盒（荧光PCR法）	20203400184	ORF1ab、N基因和E基因
11	新型冠状病毒2019-nCoV核酸检测试剂盒（荧光PCR法）	20203400058	ORF1ab和N基因
12	新型冠状病毒2019-nCoV核酸检测试剂盒（荧光PCR法）	20203400644	ORF1ab和N 基因
13	新型冠状病毒2019-nCoV核酸检测试剂盒（荧光PCR法）	20203400064	ORF1ab和N基因
14	新型冠状病毒2019-nCoV核酸检测试剂盒（荧光PCR法）	20203400063	ORF1ab和N基因
15	新型冠状病毒2019-nCoV核酸检测试剂盒（荧光PCR法）	20203400537	ORF1ab基因和N基因
16	新型冠状病毒2019-nCoV核酸检测试剂盒（荧光PCR法）	20203400179	ORF1ab、N基因和E基因
17	新型冠状病毒2019-nCoV核酸检测试剂盒（荧光PCR法）	20203400535	ORF1ab基因
18	新型冠状病毒2019-nCoV核酸检测试剂盒（荧光PCR法）	20203400212	ORF1ab基因、N基因
19	新型冠状病毒2019-nCoV核酸检测试剂盒（RNA捕获探针法）	20203400300	ORF1ab基因
20	新型冠状病毒2019-nCoV核酸检测试剂盒（RNA恒温扩增-金探针层析法）	20203400301	ORF1ab和E基因
21	新型冠状病毒2019-nCoV核酸检测试剂盒（联合探针锚定聚合测序法）	20203400059	核酸序列
22	新型冠状病毒2019-nCoV核酸检测试剂盒（恒温扩增-实时荧光法）	20203400241	ORF1ab基因、N基因

下载: 导出CSV

表 2 新型冠状病毒的国家有证核酸标准物质

Table 2. Certified nucleic acid reference materials of SARS-CoV-2

序号	标物名称	编号	标准值		定值方法
1	新型冠状病毒核酸标准物质（高浓度）	GBW(E)091089	E基因	（7.05±0.54）×10⁵ copy/μL
			ORF1ab	（1.10±0.12）×10⁶ copy/μL
			N基因	（8.38±0.76）×10⁵ copy/μL
2	新型冠状病毒核酸标准物质（低浓度）	GBW(E)091090	E基因	（7.63±0.66）×10² copy/μL
			ORF1ab	（1.24±0.19）×10³ copy/μL
			N基因	（9.8±1.2）×10² copy/μL
3	新型冠状病毒核糖核酸基因组标准物质	GBW(E)091098	E基因	（1.29±0.26）×10² copy/μL
			ORF1ab	（9.5±1.8）×10³ copy/μL
			N基因	（2.05±0.31）×10² copy/μL
4	新型冠状病毒核糖核酸基因组标准物质	GBW(E)091099	E基因	（1.06±0.11）×10³ copy/μL	数字PCR
			ORF1ab	（8.96±0.61）×10² copy/μL
			N基因	（1.73±0.13）×10³ copy/μL
5	新型冠状病毒体外转录RNA标准物质	GBW(E)091111	E基因	（2.0±0.3）×10⁵ copy/μL
			ORF1ab	（1.2±0.2）×10⁵ copy/μL
			N基因	（1.0±0.1）×10⁶ copy/μL
6	新型冠状病毒体外转录RNA标准物质	GBW(E)091112	E基因	（2.0±0.3）×10⁷ copy/μL
			ORF1ab	（1.2±0.2）×10⁷ copy/μL
			N基因	（1.0±0.1）×10⁸ copy/μL
7	新型冠状病毒（2019-nCoV）假病毒核糖核酸标准物质	NIM-RM5203	E基因	（4.6±0.9）×10² copy/μL
			ORF1ab	（6.5±1.2）×10² copy/μL
			N基因	（4.3±0.9）×10² copy/μL

下载: 导出CSV

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