用于临床新型冠状病毒核酸检测的分子诊断新技术
收稿日期: 2020-06-22
修回日期: 2020-08-19
网络出版日期: 2020-09-04
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
Received date: 2020-06-22
Revised date: 2020-08-19
Online published: 2020-09-04
Supported by
Supported by the National Natural Science Foundation of China No(61871403);Jiangsu Provincial Science Fund for Distinguished Young Scholars No(BK20180005)
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。
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 . DOI: 10.16288/j.yczz.20-189
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.
| [1] | Zhou P, Yang XL, Wang XG, Hu B, Zhang L, Zhang W, Si HR, Zhu Y, Li B, Huang CL, Chen HD, Chen J, Luo Y, Guo H, Jiang RD, Liu MQ, Chen Y, Shen XR, Wang X, Zheng XS, Zhao K, Chen QJ, Deng F, Liu LL, Yan B, Zhang FX, Wang YY, Xiao GF, Shi ZL . A pneumonia outbreak associated with a new coronavirus of probable bat origin. Nature, 2020,579(7798):270-273. |
| [2] | Wang C, Horby PW, Hayden FG, Gao GF . A novel coronavirus outbreak of global health concern. Lancet, 2020,395(10223):470-473. |
| [3] | Chan JFW, Kok KH, Zhu Z, Chu H, To KKW, Yuan SF, Yuen KY . Genomic characterization of the 2019 novel human-pathogenic coronavirus isolated from a patient with atypical pneumonia after visiting Wuhan. Emerg Microbes Infect, 2020,9(1):221-236. |
| [4] | Lu RJ, Zhao X, Li J, Niu PH, Yang B, Wu HL, Wang WL, Song H, Huang BY, Zhu N, Bi YH, Ma XJ, Zhan FX, Wang L, Hu T, Zhou H, Hu ZH, Zhou WM, Zhao L, Chen J, Meng Y, Wang J, Lin Y, Yuan JY, Xie ZH, Ma JM, Liu WJ, Wang DY, Xu WB, Holmes EC, Gao GF, Wu GZ, Chen WJ, Shi WF, Tan WJ . Genomic characterisation and epidemiology of 2019 novel coronavirus: implications for virus origins and receptor binding. Lancet, 2020,395(10224):565-574. |
| [5] | Hoffmann M, Kleine-Weber H, Schroeder S, Krüger N, Herrler T, Erichsen S, Schiergens TS, Herrler G, Wu NH, Nitsche A, Müller MA, Drosten C, P?hlmann S, . SARS- CoV-2 cell entry depends on ACE2 and TMPRSS2 and is blocked by a clinically proven protease inhibitor. Cell, 2020, 181(2): 271-280.e8. |
| [6] | Zhu N, Zhang DY, Wang WL, Li XW, Yang B, Song JD, Zhao X, Huang BY, Shi WF, Lu RJ, Niu PH, Zhan FX, Ma XJ, Wang DY, Xu WB, Wu GZ, Gao GF, Tan WJ . A novel coronavirus from patients with pneumonia in China, 2019. N Engl J Med, 2020,382(8):727-733. |
| [7] | Saiki RK, Scharf S, Faloona F, Mullis KB, Horn GT, Erlich HA, Arnheim N . Enzymatic amplification of beta- globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science, 1985,230(4732):1350-1354. |
| [8] | General Office of National Health Commission of the People's Republic of China, Office of National Administration of Traditional Chinese Medicine. Diagnosis and treatment of corona virus disease-19(7th trial edition). Chin Med, 2020,15(6):801-805. |
| [8] | 中华人民共和国国家卫生健康委员会办公厅, 国家中医药管理局办公室. 新型冠状病毒肺炎诊疗方案(试行第七版). 中国医药, 2020,15(6):801-805. |
| [9] | Xu JH, Wang SL, Zhang SX, Lai GX, Lan XP, Wu XL, Yu ZY. Methods for nucleic acid detection of 2019 novel coronavirus. Int J Lab Med, 2020, 3:1-19. |
| [9] | 许金和, 王水良, 张胜行, 赖国祥, 兰小鹏, 吴小丽, 余宗阳 . 新型冠状病毒核酸检测方法. 国际检验医学杂志, 2020,3:1-19. |
| [10] | Craighead H . Future lab-on-a-chip technologies for interrogating individual molecules. Nature, 2006,442(7101):387-393. |
| [11] | Wu JD, Dong ML, Santos S, Rigatto C, Liu Y, Lin F . Lab- on-a-chip platforms for detection of cardiovascular disease and cancer biomarkers. Sensors(Basel), 2017,17(12):2394. |
| [12] | He XP, Zou BJ, Qi XM, Chen S, Lu Y, Huang Q, Zhou GH . Methods of isothermal nucleic acid amplification-based microfluidic chips for pathogen microorganism detection. Hereditas(Beijing), 2019,41(7):611-624. |
| [12] | 何祥鹏, 邹秉杰, 齐谢敏, 陈杉, 陆妍, 黄青, 周国华 . 基于核酸等温扩增的病原微生物微流控检测技术. 遗传, 2019,41(7):611-624. |
| [13] | Yuan XF, Lv JZ, Lin XM, Zhang CY, Deng JH, Wang CX, Fan XP, Wang YG, Xu H, Wu SQ . Multiplex detection of six swine viruses on an integrated centrifugal disk using loop-mediated isothermal amplification. J Vet Diagn Invest, 2019,31(3):415-425. |
| [14] | Yan H, Zhu YZ, Zhang Y, Wang L, Chen JG, Lu Y, Xu YC, Xing WL . Multiplex detection of bacteria on an integrated centrifugal disk using bead-beating lysis and loop-mediated amplification. Sci Rep, 2017,7(1):1460. |
| [15] | 刘伟 . 基于RNA靶标SAT解脲脲原体感染分子生物学检测方法. CN108611403A, 2018-10-02. |
| [16] | 高通量、全流程一体化的新冠核酸检测平台. , 2020-03-29. |
| [17] | Fang RD, Li X, Hu L, You QM, Li J, Wu J, Xu P, Zhong HY, Luo Y, Mei J, Gao Q . Cross-priming amplification for rapid detection of mycobacterium tuberculosis in sputum specimens. J Clin Microbiol, 2009,47(3):845-847. |
| [18] | 杭州一企业紧急研制新冠病毒核酸检测试剂盒最快30分钟出结果. , 2020-01-31. |
| [19] | You QM . Cross priming amplification of target nucleic acids. CA2748822, 2016 -09-06-18. |
| [20] | 祁军, 尤其敏, 柴宏森, 左锋, 王馨, 詹曦菁, 刘振宇, 刘寅, 杨春江, 刘智勇, 徐高连, 刘启军, 张霞, 崔景柏, 王宏莹, 高秋萍, 吴汀滢 . 运用交叉引物核酸恒温扩增技术检测肠出血性大肠埃希菌的试剂及其扩增方法和检测方法. CN102363816A, 2012-02-29. |
| [21] | 汪大明, 钟乾兴, 胡啸, 张利伟, 肖江群, 王保丹, 江应玲, 乐宜萃 . 一种核酸杂交捕获免疫荧光检测方法、免疫荧光层析试条及试剂盒. CN109613236A, 2019-04-12. |
| [22] | 新型冠状病毒核酸-安邦(厦门)生物科技有限公司. , 2020-03. |
| [23] | 一文读懂测序技术在新冠病毒检测中的应用. , 2020-03-27. |
| [24] | Guo Z, Wang F, Guo YX, Liu XW, Yang XG, Song YF, Wang DG . Potential value of droplet digital PCR in detection of SARS-CoV-2. J Lanzhou Univ(Med Sci). 2020,46(02):14-18. |
| [24] | 郭兆, 王芳, 郭妍譞, 刘向文, 杨旭光, 宋焱峰, 王德贵 . 微滴式数字PCR技术在新型冠状病毒检测中的潜在价值. 兰州大学学报(医学版), 2020,46(2):14-18. |
| [25] | Yu FT, Yan LT, Wang N, Yang SY, Wang LH, Tang YX, Gao GJ, Wang S, Ma CJ, Xie RM, Wang F, Tan C, Zhu LX, Guo Y, Zhang FJ . Quantitative detection and viral load analysis of SARS-CoV-2 in infected patients. Clin Infect Dis, 2020,71(15):793-798. |
| [26] | Wahed AAE, Patel P, Heidenreich D, Hufert FT, Weidmann M . Reverse transcription recombinase polymerase amplification assay for the detection of middle east respiratory syndrome coronavirus. PLoS Curr, 2013,5(8):813-818. |
| [27] | Gootenberg JS, Abudayyeh OO, Lee JW, Essletzbichler P, Dy AJ, Joung J, Verdine V, Donghia N, Daringer NM, Freije CA, Myhrvold C, Bhattacharyya RP, Livny J, Regev A, Koonin EV, Hung DT, Sabeti PC, Collins JJ, Zhang F . Nucleic acid detection with CRISPR-Cas13a/C2c2. Science, 2017,356(6336):438-442. |
| [28] | Wang XJ, Zhong MT, Liu Y, Ma PX, Dang L, Meng QZ, Wan WW, Ma XD, Liu J, Yang G, Yang ZF, Huang XX, Liu M . Rapid and sensitive detection of COVID-19 using CRISPR/Cas12a-based detection with naked eye readout, CRISPR/Cas12a-NER. Sci Bull(Beijing), 2020,65(17):1436-1439. |
| [29] | Enabling coronavirus detection using CRISPR-Cas13: Open-access SHERLOCK research protocols and design resources | Broad Institute. |
/
| 〈 |
|
〉 |
www.chinagene.cn
备案号：京ICP备09063187号-4
总访问:,今日访问:,当前在线:
