2020年 第6卷 第10期
《工程（英文）》 >> 2020年 第6卷 第10期 doi: 10.1016/j.eng.2020.07.015
一款用于快速检测包括SARS-CoV-2 在内的19 种常见呼吸道病毒的高通量、多指标等温扩增平台
a Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084, China
b National Engineering Research Center for Beijing Biochip Technology, Beijing 102206, China
c CapitalBio Technology, Beijing 101111, China
d CapitalBio Corporation, Beijing 102206, China
e Department of Basic Medical Sciences, School of Medicine, Tsinghua University, Beijing 100084, China
f Department of Laboratory Medicine, First Affiliated Hospital of Guangzhou Medical University, Guangzhou 510120, China
g Department of Neurology, West China Hospital, Sichuan University, Chengdu 610041, China
h Clinical Laboratory Center, Beijing Youan Hospital, Capital Medical University, Beijing 100069, China
i Department of Pediatrics, Beijing Tsinghua Changgung Hospital, School of Clinical Medicine, Tsinghua University, Beijing 102218, China
j Experiment Center, Capital Institute of Pediatrics, Beijing 100020, China
k President's Office, Beijing Youan Hospital, Capital Medical University, Beijing 100069, China
l Department of Cardiothoracic Surgery, State Key Laboratory of Respiratory Disease, China Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, First Affiliated Hospital of Guangzhou Medical University, Guangzhou 510120, China
m Department of Clinical Laboratory, Beijing Tsinghua Changgung Hospital, School of Clinical Medicine, Tsinghua University, Beijing 102218, China
n Department of Respiratory and Critical Care Medicine, West China Hospital, Sichuan University, Chengdu 610041, China
# These authors contributed equally to this work.
下一篇 上一篇
摘要
快速、准确地诊断以及及时对感染严重急性呼吸系统综合征冠状病毒2（SARS-CoV-2）的患者进行隔离被认为是抗击新冠病毒肺炎（COVID-19）大流行的最有效的措施。本文介绍了一款高通量、多指标的等温扩增检测平台（RTisochipTM-W），该平台使用离心式微流控芯片，在90 min内即可完成一次运行，从16个样本中检测出包括SARS-CoV-2在内的19种常见呼吸道病毒。通过RTisochipTM-W系统检测的所有病毒的检出限均等于或小于50拷贝· μL−1，与常规逆转录聚合酶链反应（RT-PCR）灵敏度相当。该系统具有重复性好、鲁棒性强和特异性高的优点。最后，使用RTisochipTM-W系统对201例临床前样本、14例COVID-19阳性临床样本、25例临床确诊样本以及614例来自呼吸道感染或疑似感染的患者的临床样本进行了测试。测试结果与临床诊断结果吻合良好，也反映出呼吸道传染病的流行特征。在对SARS-CoV-2的诊断上，RTisochipTM-W系统与参考试剂盒的一致性达到98.15%。基于大量的测试结果，我们相信RTisochipTM-W系统是一款抗击COVID-19大流行的强有力工具。
关键词
COVID-19 ; SARS-CoV-2 ; 微流控 ; 核酸检测 ; 等温扩增
补充材料
图片
图1
图2
图3
图4
图5
图6
图7
参考文献
[ 1 ] Wang C, Horby PW, Hayden FG, Gao GF. A novel coronavirus outbreak of global health concern. Lancet 2020;395(10223):470–3. 链接1
[ 2 ] Huang C, Wang Y, Li X, Ren L, Zhao J, Hu Y, et al. Clinical features of patients infected with 2019 novel coronavirus in Wuhan, China. Lancet 2020;395 (10223):497–506. 链接1
[ 3 ] Corman VM, Landt O, Kaiser M, Molenkamp R, Meijer A, Chu DKW, et al. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Euro Surveill 2020;25(3):2000045. 链接1
[ 4 ] Chu DKW, Pan Y, Cheng SMS, Hui KPY, Krishnan P, Liu Y, et al. Molecular diagnosis of a novel coronavirus (2019-nCoV) causing an outbreak of pneumonia. Clin Chem 2020;66(4):549–55. 链接1
[ 5 ] Wilder-Smith A, Freedman DO. Isolation, quarantine, social distancing and community containment: pivotal role for old-style public health measures in the novel coronavirus (2019-nCoV) outbreak. J Travel Med 2020;27 (2):taaa020.
[ 6 ] Li G, De Clercq E. Therapeutic options for the 2019 novel coronavirus (2019- nCoV). Nat Rev Drug Discov 2020;19(3):149–50. 链接1
[ 7 ] Zhai P, Ding Y, Wu X, Long J, Zhong Y, Li Y. The epidemiology, diagnosis and treatment of COVID-19. Int J Antimicrob Agents 2020;55(5):105955. 链接1
[ 8 ] Westh H, Lisby G, Breysse F, Böddinghaus B, Chomarat M, Gant V, et al. Multiplex real-time PCR and blood culture for identification of bloodstream pathogens in patients with suspected sepsis. Clin Microbiol Infect 2009;15 (6):544–51. 链接1
[ 9 ] Li Y, Guo SJ, Shao N, Tu S, Xu M, Ren ZR, et al. A universal multiplex PCR strategy for 100-plex amplification using a hydrophobically patterned microarray. Lab Chip 2011;11(21):3609–18. 链接1
[10] Wang Y, Sims CE, Allbritton NL. Dissolution-guided wetting for microarray and microfluidic devices. Lab Chip 2012;12(17):3036–9. 链接1
[11] Du W, Li L, Nichols KP, Ismagilov RF. SlipChip. Lab Chip 2009;9(16):2286–92. 链接1
[12] Shen C, Xu P, Huang Z, Cai D, Liu SJ, Du W. Bacterial chemotaxis on SlipChip. Lab Chip 2014;14(16):3074–80. 链接1
[13] Shen F, Du W, Davydova EK, Karymov MA, Pandey J, Ismagilov RF. Nanoliter multiplex PCR arrays on a SlipChip. Anal Chem 2010;82(11):4606–12. 链接1
[14] Zhukov DV, Khorosheva EM, Khazaei T, Du W, Selck DA, Shishkin AA, et al. Microfluidic SlipChip device for multistep multiplexed biochemistry on a nanoliter scale. Lab Chip 2019;19(19):3200–11. 链接1
[15] Barbulovic-Nad I, Yang H, Park PS, Wheeler AR. Digital microfluidics for cellbased assays. Lab Chip 2008;8(4):519–26. 链接1
[16] Sista R, Hua Z, Thwar P, Sudarsan A, Srinivasan V, Eckhardt A, et al. Development of a digital microfluidic platform for point of care testing. Lab Chip 2008;8(12):2091–104. 链接1
[17] Abd Rahman N, Ibrahim F, Aeinehvand MM, Yusof R, Madou M. A microfluidic Lab-on-a-Disc (LOD) for antioxidant activities of plant extracts. Micromachines 2018;9(4):140. 链接1
[18] Stumpf F, Schwemmer F, Hutzenlaub T, Baumann D, Strohmeier O, Dingemanns G, et al. LabDisk with complete reagent prestorage for sampleto-answer nucleic acid based detection of respiratory pathogens verified with influenza A H3N2 virus. Lab Chip 2016;16(1):199–207. 链接1
[19] Hou J, Wu H, Zeng X, Rao H, Zhao P. Clinical evaluation of the loop-mediated isothermal amplification assay for the detection of common lower respiratory pathogens in patients with respiratory symptoms. Medicine (Baltimore) 2018;97(51):e13660. 链接1
[20] Hanson KE, Couturier MR. Multiplexed molecular diagnostics for respiratory, gastrointestinal, and central nervous system infections. Clin Infect Dis 2016;63 (10):1361–7. 链接1
[21] Popowitch EB, O’Neill SS, Miller MB. Comparison of the Biofire FilmArray RP, Genmark eSensor RVP, Luminex xTAG RVPv1, and Luminex xTAG RVP fast multiplex assays for detection of respiratory viruses. J Clin Microbiol 2013;51 (5):1528–33. 链接1
[22] Rand KH, Rampersaud H, Houck HJ. Comparison of two multiplex methods for detection of respiratory viruses: FilmArray RP and xTAG RVP. J Clin Microbiol 2011;49(7):2449–53. 链接1
[23] Compton J. Nucleic acid sequence-based amplification. Nature 1991;350 (6313):91–2. 链接1
[24] Malek L, Sooknanan R, Compton J. Nucleic acid sequence-based amplification (NASBA). Methods Mol Biol 1994;28:253–60. 链接1
[25] Van Gemen B, Kievits T, Nara P, Huisman HG, Jurriaans S, Goudsmit J, et al. Qualitative and quantitative detection of HIV-1 RNA by nucleic acid sequencebased amplification. AIDS 1993;7(Suppl 2):S107–10. 链接1
[26] Mugasa CM, Katiti D, Boobo A, Lubega GW, Schallig HDFH, Matovu E. Comparison of nucleic acid sequence-based amplification and loop-mediated isothermal amplification for diagnosis of human African trypanosomiasis. Diagn Microbiol Infect Dis 2014;78(2):144–8. 链接1
[27] Mercier-Delarue S, Vray M, Plantier JC, Maillard T, Adjout Z, de Olivera F, et al. Higher specificity of nucleic acid sequence-based amplification isothermal technology than of real-time PCR for quantification of HIV-1 RNA on dried blood spots. J Clin Microbiol 2014;52(1):52–6. 链接1
[28] Dimov IK, Garcia-Cordero JL, O’Grady J, Poulsen CR, Viguier C, Kent L, et al. Integrated microfluidic tmRNA purification and real-time NASBA device for molecular diagnostics. Lab Chip 2008;8(12):2071–8. 链接1
[29] Houde A, Leblanc D, Poitras E, Ward P, Brassard J, Simard C, et al. Comparative evaluation of RT-PCR, nucleic acid sequence-based amplification (NASBA) and real-time RT-PCR for detection of noroviruses in faecal material. J Virol Methods 2006;135(2):163–72. 链接1
[30] Huang G, Huang Q, Xie L, Xiang G, Wang L, Xu H, et al. A rapid, low-cost, and microfluidic chip-based system for parallel identification of multiple pathogens related to clinical pneumonia. Sci Rep 2017;7:6441. 链接1
[31] Zhang G, Zheng G, Zhang Y, Ma R, Kang X. Evaluation of a micro/nanofluidic chip platform for the high-throughput detection of bacteria and their antibiotic resistance genes in post-neurosurgical meningitis. Int J Infect Dis 2018;70:115–20. 链接1
[32] Yu F, Yan L,Wang N, Yang S,Wang L, Tang Y, et al. Quantitative detection and viral load analysis of SARS-CoV-2 in infected patients. Clin Infect Dis 2020. In press. 链接1
[33] Kim JY, Ko JH, Kim Y, Kim YJ, Kim JM, Chung YS, et al. Viral load kinetics of SARS-CoV-2 infection in first two patients in Korea. J Korean Med Sci 2020;35 (7):e86. 链接1
[34] Liu Y, Yang Y, Zhang C, Huang F, Wang F, Yuan J, et al. Clinical and biochemical indexes from 2019-nCoV infected patients linked to viral loads and lung injury. Sci China Life Sci 2020;63(3):364–74. 链接1
[35] National Health Commission of the People’s Republic of China, National Administration of Traditional Chinese Medicine. Diagnosis and treatment protocol for novel coronavirus pneumonia (trial version 7). Chin Med J Epub 2020;133(9):1087–95.
京公网安备 11010502051620号
