核酸检测技术在SARS-CoV-2检测中研究进展

柴瑜; 唐继海

doi:10.19837/j.cnki.ahyf.2021.03.014

PDF(1153 KB)

安徽预防医学杂志 ›› 2021, Vol. 27 ›› Issue (3) : 223-228. DOI: 10.19837/j.cnki.ahyf.2021.03.014

综述

核酸检测技术在SARS-CoV-2检测中研究进展

柴瑜, 唐继海

作者信息 +

Research progress of nucleic acid detection technology for SARS-CoV-2

CHAI Yu, TANG Ji-hai

Author information +

文章历史 +

摘要

严重急性呼吸综合征冠状病毒2(SARS-CoV-2)给人类健康带来严重威胁,由此引发的新型冠状病毒肺炎疫情席卷全球,因此快速、准确地检测技术是早期发现疫情的关键手段,对有效控制疫情的蔓延起到重要作用。核酸检测因检测速度快、准确性好、灵敏度高,已经成为目前疾病检测和筛查最常用的方法。本文将综述荧光定量PCR、恒温核酸扩增、数字PCR 和测序等方法的原理以及在新冠病毒检测中的应用进展,希望能为SARS-CoV-2检测和科研工作提供参考。

Abstract

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) posed a serious threat to human health,and the resulting Corona Virus Disease 2019 (COVID-19) had swept the world.Therefore,rapid and accurate detection technology was a key means for early detection of the epidemic,which played an important role in effectively controlling the spread of the epidemic.Nucleic acid detection had become the most commonly used method for disease detection and screening due to its fast detection speed,good accuracy,and high sensitivity.This article will review the principles of Realtime PCR,loop-mediated isothermal amplification,digital PCR,and sequencing,as well as the application progress in the detection of COVID-19,hoping to provide a reference for SARS-CoV-2 detection and research.

导出引用

柴瑜, 唐继海. 核酸检测技术在SARS-CoV-2检测中研究进展[J]. 安徽预防医学杂志. 2021, 27(3): 223-228 https://doi.org/10.19837/j.cnki.ahyf.2021.03.014

CHAI Yu, TANG Ji-hai. Research progress of nucleic acid detection technology for SARS-CoV-2[J]. Anhui Journal of Preventive Medicine. 2021, 27(3): 223-228 https://doi.org/10.19837/j.cnki.ahyf.2021.03.014

中图分类号： R373

参考文献

[1] Gorbalenya A E,Baker S C,Baric R S,et al.Severe acute respiratory syndrome-related coronavirus: The species and its viruses – a statement of the Coronavirus Study Group.2020.
[2] 国家卫生健康委,新型冠状病毒肺炎防控方案(第七版).2020.
[3] Wan Y,Shang J,Graham R,et al.Receptor Recognition by the Novel Coronavirus from Wuhan: an Analysis Based on Decade-Long Structural Studies of SARS Coronavirus[J].J Virol,2020,94(7).
[4] Chan JF,Kok KH,Zhu Z,et al.Genomic characterization of the 2019 novel human-pathogenic coronavirus isolated from a patient with atypical pneumonia after visiting Wuhan[J].Emerg Microbes Infect,2020,9(1): 221-236.
[5] Cai Y,Zhang J,Xiao T,et al.Distinct conformational states of SARS-CoV-2 spike protein[J].bioRxiv,2020.
[6] WHO.WHO Coronavirus Disease (COVID-19) Dashboard Data last updated: 2021/2/9.Available from: https://covid19.who.int/.
[7] 郭金英.初探不同标本核酸检测在新型冠状病毒肺炎患者诊断中的应用[J].长治医学院学报,2020,2(34): 97-100.
[8] 陈炜,张春阳,朱颖,等.4例新型冠状病毒感染病例咽拭子与痰标本病毒核酸检测的比较[J].中国人兽共患病学报,2020,5(36): 354-358.
[9] 国家卫生健康委,新型冠状病毒肺炎防控方案(第七版).2020.
[10] 刘焱斌,刘涛,崔跃,等.鼻拭子与咽拭子两种取样方法在新型冠状病毒肺炎核酸筛检中的比较研究[J].中国呼吸与危重监护杂志,2020,19(2):141-143.
[11] 美国疾病预防控制中心,2019-Novel Coronavirus (2019-nCoV) Real-time rRT-PCR Panel Primers and Probes.2020.
[12] 国家卫生健康委,新型冠状病毒肺炎防控方案(第二版).2020.
[13] 孟影,陈春峰,张小燕,等.2019新型冠状病毒核酸检测试剂盒的研发与应用[J].中国医药生物技术,2020,3(15): 230-234.
[14] Corman VM,Landt O,Kaiser M,et al.Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR[J].Euro Surveill,2020,25(3).
[15] Notomi T,Okayama H,Masubuchi H,et al.Loop-mediated isothermal amplification of DNA[J].Nucleic Acids Res,2000,28(12): E63.
[16] 关丽,聂凯,张丹,等.环介导逆转录等温扩增技术检测中东呼吸综合征冠状病毒基因[J].病毒学报,2015,3(31): 269-275.
[17] 吕恒,张锦海,陈凤娟,等.SARS冠状病毒环介导等温扩增可视化检测方法的建立[J].实用预防医学,2013,4(20): 395-399.
[18] 耿合员,汪圣强,谢晓谦,等.基于颜色判定的RT-LAMP技术检测HCoV-NL63冠状病毒基因[J].病毒学报,2016,32(1): 56-61.
[19] 耿合员,汪圣强,吴海磊,等.基于颜色判定的RT-LAMP技术检测HCoV-OC43冠状病毒[J].中国国境卫生检疫杂志,2018,41(3): 153-158.
[20] 刘胜牙,朱玉兰,胡孔新,等.人冠状病毒HCoV-HKU1环介导等温扩增检测技术的研究[J].中国国境卫生检疫杂志,2014,37(4): 217-220.
[21] Lamb LE,Bartolone SN,Ward E,et al.Rapid detection of novel coronavirus/Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) by reverse transcription-loop-mediated isothermal amplification[J].PLoS One,2020,15(6): e0234682.
[22] Yu L,Wu S,Hao X,et al.Rapid Detection of COVID-19 Coronavirus Using a Reverse Transcriptional Loop-Mediated Isothermal Amplification (RT-LAMP) Diagnostic Platform[J].Clin Chem,2020,66(7): 975-977.
[23] Zhu X,Wang X,Han L,et al.Multiplex reverse transcription loop-mediated isothermal amplification combined with nanoparticle-based lateral flow biosensor for the diagnosis of COVID-19[J].Biosens Bioelectron,2020,166: 112437.
[24] Ding S,Chen G,Wei Y,et al.Sequence-specific and multiplex detection of COVID-19 virus (SARS-CoV-2) using proofreading enzyme-mediated probe cleavage coupled with isothermal amplification[J].Biosens Bioelectron,2021,178: 113041.
[25] Sykes PJ,Neoh SH,Brisco MJ,et al.Quantitation of targets for PCR by use of limiting dilution[J].Biotechniques,1992,13(3): 444-449.
[26] Vogelstein B,Kinzler KW.Digital PCR[J].Proc Natl Acad Sci U S A,1999,96(16): 9236-9241.
[27] Dressman D,Yan H,Traverso G,et al.Transforming single DNA molecules into fluorescent magnetic particles for detection and enumeration of genetic variations[J].Proc Natl Acad Sci U S A,2003,100(15): 8817-8822.
[28] 冯秀晶,易红梅,任星旭,等.数字PCR技术及其在检测领域的应用[J].遗传,2020,42(4): 363-373.
[29] 李彤,荆福祥,李宏志,等.新型冠状病毒数字PCR检测方法建立[J].分子诊断与治疗杂志,2020,12(7): 843-847.
[30] Ahmed W,Bertsch PM,Angel N,et al.Detection of SARS-CoV-2 RNA in commercial passenger aircraft and cruise ship wastewater: a surveillance tool for assessing the presence of COVID-19 infected travellers[J].J Travel Med,2020,27(5).
[31] Lv J,Yang J,Xue J,et al.Detection of SARS-CoV-2 RNA residue on object surfaces in nucleic acid testing laboratory using droplet digital PCR[J].Sci Total Environ,2020,742: 140370.
[32] Suo T,Liu X,Feng J,et al.ddPCR: a more accurate tool for SARS-CoV-2 detection in low viral load specimens[J].Emerg Microbes Infect,2020,9(1): 1259-1268.
[33] Yu F,Yan L,Wang N,et al.Quantitative Detection and Viral Load Analysis of SARS-CoV-2 in Infected Patients[J].Clin Infect Dis,2020,71(15): 793-798.
[34] Ai JW,Zhang Y,Zhang HC,et al.Era of molecular diagnosis for pathogen identification of unexplained pneumonia,lessons to be learned[J].Emerg Microbes Infect,2020,9(1): 597-600.
[35] Manning JE,Bohl JA,Lay S,et al.Rapid metagenomic characterization of a case of imported COVID-19 in Cambodia[J].bioRxiv,2020.
[36] 李夫,潘阳,王雅杰,等.北京市12例境外输入新冠肺炎病例病毒基因组特征分析[J].国际病毒学杂志,2020,27(3): 177-181.
[37] 梁丽君,李柏生,刘哲,等.广东省新型冠状病毒全基因组序列测定及其影响因素分析[J].中华微生物学和免疫学杂志,2020,(3): 174-177.
[38] 耿合员,田锋,朱娜,等.基于毛细管电泳的快速检测七种人冠状病毒技术方法的建立[J].病毒学报,2021,37(1): 25-33.
[39] 林长缨,丁晓静.毛细管电泳技术在疾病预防控制领域的应用、发展与挑战[J].色谱,2020,38(9): 999-1012.
[40] Metzker ML.Sequencing technologies-the next generation[J].Nat Rev Genet,2010,11(1): 31-46.
[41] 邹明园,吴国球.抗原交叉反应对新型冠状病毒血清特异性抗体检测的影响[J].临床检验杂志,2020,3(38): 161-163.
[42] 徐万洲,李娟,何晓云,等.血清2019新型冠状病毒IgM和IgG抗体联合检测在新型冠状病毒感染中的诊断价值[J].中华检验医学杂志,2020,43(3):230-233.
[43] 雷松菁,王刚平,周刚,等.新型冠状病毒SARS-CoV-2基因检测及其影像学对照[J].分子诊断与治疗杂志,2020,12(3): 270-274.
[44] 邱世香,魏剑林,胡鸿,等.新型冠状病毒肺炎的CT检查与核酸检测对比[J].西部医学,2020,32(5): 638-642.
[45] 张新,郭伟,刘晓倩,等.反复病毒核酸检测阴性的疑似新型冠状病毒肺炎一例[J].中华临床感染病杂志,2020,13(1): 39-41.
[46] 李园园,王伟娜,雷宇,等.临床诊断新型冠状病毒肺炎中核酸检测阳性与阴性患者临床特征的比较[J].中华结核和呼吸杂志,2020,43(5): 427-430.
[47] 李晓非,刘贵明,李婉澜,等.化学发光法检测新型冠状病毒抗体在临床的应用价值[J].山东大学学报(医学版),2020,58(5): 46-50.
[48] 马红霞,潘静静,李懿,等.实时荧光RT-PCR方法检测新型冠状病毒核酸结果分析[J].中华微生物学和免疫学杂志,2020,(4): 245-249.