用于临床新型冠状病毒核酸检测的分子诊断新技术

doi:10.16288/j.yczz.20-189

遗传 ›› 2020, Vol. 42 ›› Issue (9): 870-881.doi: 10.16288/j.yczz.20-189

用于临床新型冠状病毒核酸检测的分子诊断新技术

谢春梅¹, 武海萍¹, 马雪萍¹, 周国华^1,²()

^1. 南京大学医学院附属金陵医院(东部战区总医院)临床药学科,南京 210002
^2. 南方医科大学药学院,广州 510515

收稿日期:2020-06-22 修回日期:2020-08-19 出版日期:2020-09-20 发布日期:2020-09-04
通讯作者: 周国华 E-mail:ghzhou@nju.edu.cn
基金资助:
国家自然科学基金项目编号(61871403);江苏省杰出青年基金项目资助编号(BK20180005)

New molecular diagnostic technologies for clinical detection of SARS-CoV-2

Chunmei Xie¹, Haiping Wu¹, Xueping Ma¹, Guohua Zhou^1,²()

^1. Department of Clinical Pharmacy, Jinling Hospital, Medical School of Nanjing University, Nanjing 210002, China
^2. School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, China

Received:2020-06-22 Revised:2020-08-19 Online:2020-09-20 Published:2020-09-04
Contact: Zhou Guohua E-mail:ghzhou@nju.edu.cn
Supported by:
Supported by the National Natural Science Foundation of China No(61871403);Jiangsu Provincial Science Fund for Distinguished Young Scholars No(BK20180005)

摘要/Abstract

摘要：

目前新型冠状病毒肺炎(corona virus disease 2019, COVID-19)在全球仍呈现大流行趋势,该病潜伏期长、传染性强,给全世界人民的健康安全带来严重威胁。新型冠状病毒(severe acute respiratory syndrome coronavirus 2, SARS-CoV-2)核酸检测作为COVID-19确诊的重要依据之一,在疫情防控工作中发挥了巨大的作用。截至2020年8月17日,国家药品监督管理局已应急批准15个新型冠状病毒核酸检测试剂盒,其中10个试剂盒是以逆转录-实时荧光定量PCR (RT-qPCR)为主要技术原理,其余试剂盒分别采用了不同于RT-qPCR的5种分子诊断技术。本文对上述检测试剂盒所使用的技术原理、反应时间、优缺点等方面进行了介绍,以期为COVID-19及类似传染病的快速筛查、确诊、防控提供更多的方案思路。

关键词: 新型冠状病毒, 核酸检测, 分子诊断

Abstract:

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused an ongoing pandemic of new coronavirus pneumonia (corona virus disease 2019, COVID-19). The virus has a long incubation period and strong infectivity, which poses a major threat to global health and safety. Detection of SARS-CoV-2 nucleic acid lies at the center of rapid detection of COVID-19, which is instrumental for mitigation of the ongoing pandemic. As of August 17, 2020, The National Medical Products Administration in China has approved 15 new coronavirus nucleic acid detection kits, 10 kits of which are based on reverse transcription-real-time quantitative PCR (RT-qPCR) technology. The remaining kits use five molecular diagnostic technologies different from RT-qPCR. This article reviews the principles, reaction time, advantages and disadvantages of above 15 detection kits, in order to provide references for rapid screening, diagnosis, prevention and control of COVID-19 and similar infectious diseases.

Key words: novel coronavirus, nucleic acid detection, molecular diagnostic technology

谢春梅, 武海萍, 马雪萍, 周国华. 用于临床新型冠状病毒核酸检测的分子诊断新技术[J]. 遗传, 2020, 42(9): 870-881.

Chunmei Xie, Haiping Wu, Xueping Ma, Guohua Zhou. New molecular diagnostic technologies for clinical detection of SARS-CoV-2[J]. Hereditas(Beijing), 2020, 42(9): 870-881.

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试剂盒名称	公司	分子诊断技术	扩增方式	检测方式	检测靶标
6项呼吸道病毒核酸检测试剂盒(恒温扩增芯片法)	成都博奥晶芯生物科技有限公司	恒温扩增芯片技术	环介导等温扩增	荧光检测	S、N
新型冠状病毒2019-nCoV核酸检测试剂盒(RNA捕获探针法)	上海仁度生物科技有限公司	SAT-RNA实时荧光恒温扩增技术	T7 RNA聚合酶扩增	荧光检测	ORFlab
新型冠状病毒2019-nCoV核酸检测试剂盒(恒温扩增-实时荧光法)	杭州优思达生物技术有限公司	交叉引物恒温扩增——实时荧光技术	交叉引物恒温扩增	荧光检测	ORF1ab、N
新型冠状病毒2019-nCoV核酸检测试剂盒(杂交捕获免疫荧光法)	安邦(厦门)生物科技有限公司	杂交捕获免疫荧光技术	无需扩增	试纸条可视化判读	ORF1ab、N、E
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)	华大生物科技(武汉)有限公司	联合探针锚定聚合测序技术	DNA纳米球扩增	光信号识别	核酸序列

分子诊断技术	反应时间(min)	检测样本类型	检测限	临床灵敏度(%)	临床特异性(%)	优点	缺点
RT-qPCR技术	120~180	肺泡灌洗液、咽拭子等样本	100~300拷贝/mL	100	99	高灵敏、高特异、可定量	耗时长、存在假阳性及假阴性问题
恒温扩增芯片技术	90	鼻拭子、咽拭子样本	15拷贝/反应	-	-	可同时检测多个靶标	未完全实现自动化
SAT-RNA实时荧光恒温扩增技术	90	咽拭子、痰液样本	-	-	-	全流程自动化、高通量	价格昂贵、循环反应过程复杂
交叉引物恒温扩增——实时荧光技术	30	咽拭子、痰液、血清、血浆、粪便样本	10拷贝/反应	100	100	全流程自动化、操作简便	单次只能检测2个样本
杂交捕获免疫荧光技术	45	咽拭子、痰液样本	500 拷贝/mL	88.61	94.92	无需提取纯化、无需核酸扩增、实现一步现场快速检测	无法定量、灵敏度较低
联合探针锚定聚合测序技术	60	鼻拭子、咽拭子样本	-	-	-	快速高效、准确度高、通量大、可监测病毒突变	结果易受环境温度、检测试剂的影响

用于临床新型冠状病毒核酸检测的分子诊断新技术

New molecular diagnostic technologies for clinical detection of SARS-CoV-2

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