Direct on-the-spot detection of SARS-CoV-2 in patients

被引:31
作者
Ben-Assa, Nadav [1 ]
Naddaf, Rawi [1 ]
Gefen, Tal [1 ]
Capucha, Tal [1 ,2 ]
Hajjo, Haitham [1 ,3 ,4 ]
Mandelbaum, Noa [1 ]
Elbaum, Lilach [1 ]
Rogov, Peter
King, Daniel A. [5 ]
Kaplan, Shai [6 ]
Rotem, Assaf
Chowers, Michal [7 ,8 ]
Szwarcwort-Cohen, Moran [9 ]
Paul, Mical [10 ,11 ]
Geva-Zatorsky, Naama [1 ,12 ]
机构
[1] Technion Israel Inst Technol, Technion Integrated Canc Ctr TICC, Rappaport Fac Med, Dept Cell Biol & Canc Sci, IL-3525433 Haifa, Israel
[2] Rambam Med Care Ctr, Dept Oral & Maxillofacial Surg, IL-31999 Haifa, Israel
[3] Technion Israel Inst Technol, Rappaport Fac Med, Dept Immunol, IL-3525433 Haifa, Israel
[4] Technion Israel Inst Technol, Rappaport Fac Med, Dept Neurosci, IL-3525433 Haifa, Israel
[5] Meir Med Ctr, Pulm & Resp Crit Care Div, IL-44281 Kefar Sava, Israel
[6] Robiotec Ltd, IL-7634709 Rehovot, Israel
[7] Meir Med Ctr, Infect Dis Unit, IL-44281 Kefar Sava, Israel
[8] Tel Aviv Univ, Sackler Fac Med, IL-3200 Tel Aviv, Israel
[9] Rambam Hlth Care Campus, Virol Lab, IL-31999 Haifa, Israel
[10] Technion Israel Inst Technol, Div Infect Dis, Rambam Hlth Care Ctr, IL-31999 Haifa, Israel
[11] Technion Israel Inst Technol, Rappaport Fac Med, IL-31999 Haifa, Israel
[12] Canadian Inst Adv Res CIFAR Azrieli Global Schola, MaRS Ctr, Toronto, ON M5G 1M1, Canada
关键词
Molecular; Covid-19; SARS-CoV-2; RT-LAMP; surveillance; pandemic;
D O I
10.1177/1535370220941819
中图分类号
R-3 [医学研究方法]; R3 [基础医学];
学科分类号
1001 ;
摘要
Many countries are currently in a state of lockdown due to the SARS-CoV-2 pandemic. One key requirement to safely transition out of lockdown is the continuous testing of the population to identify infected subjects. Currently, detection is performed at points of care using quantitative reverse-transcription PCR, thus requiring dedicated professionals and equipment. Here, we developed a protocol based on reverse transcribed loop-mediated isothermal amplification for the detection of SARS-CoV-2. This protocol is applied directly to SARS-CoV-2 nose and throat swabs, with no RNA purification step required. We tested this protocol on over 180 suspected patients, and compared the results to those obtained using the standard method. We further succeeded in applying the protocol to self-collected saliva samples from confirmed cases. Since the proposed protocol can detect SARS-CoV-2 from saliva and provides on-the-spot results, it allows simple and continuous surveillance of the community. Impact statement Humanity is currently experiencing a global pandemic with devastating implications on human health and the economy. Most countries are gradually exiting their lockdown state. We are currently lacking rapid and simple viral detections, especially methods that can be performed in the household. Here, we applied RT-LAMP directly on human clinical swabs and self-collected saliva samples. We adjusted the method to allow simple and rapid viral detection, with no RNA purification steps. By testing our method on over 180 human samples, we determined its sensitivity, and by applying it to other viruses, we determined its specificity. We believe this method has a promising potential to be applied world-wide as a simple and cheap surveillance test for SARS-CoV-2.
引用
收藏
页码:1187 / 1193
页数:7
相关论文
共 11 条
  • [1] [Anonymous], 2020, Clin Infect Dis, DOI DOI 10.1093/CID/CIAA797
  • [2] Bruce Emily A, 2020, PLoS Biol, V18, pe3000896, DOI [10.1101/2020.03.20.001008, 10.1371/journal.pbio.3000896]
  • [3] A multiplexed RT-LAMP assay for detection of group M HIV-1 in plasma or whole blood
    Curtis, Kelly A.
    Morrison, Daphne
    Rudolph, Donna L.
    Shankar, Anupama
    Bloomfield, Laura S. P.
    Switzer, William M.
    Owen, S. Michele
    [J]. JOURNAL OF VIROLOGICAL METHODS, 2018, 255 : 91 - 97
  • [4] Lassaunie` re R, 2020, MEDRXIV, DOI 10.1101/2020.04.09.20056325
  • [5] Droplets and Aerosols in the Transmission of SARS-CoV-2
    Meselson, Matthew
    [J]. NEW ENGLAND JOURNAL OF MEDICINE, 2020, 382 (21) : 2063 - 2063
  • [6] Loop-mediated isothermal amplification of DNA
    Notomi, Tsugunori
    Okayama, Hiroto
    Masubuchi, Harumi
    Yonekawa, Toshihiro
    Watanabe, Keiko
    Amino, Nobuyuki
    Hase, Tetsu
    [J]. NUCLEIC ACIDS RESEARCH, 2000, 28 (12)
  • [7] Laboratory Diagnosis of COVID-19: Current Issues and Challenges
    Tang, Yi-Wei
    Schmitz, Jonathan E.
    Persing, David H.
    Stratton, Charles W.
    [J]. JOURNAL OF CLINICAL MICROBIOLOGY, 2020, 58 (06)
  • [8] Visual detection of isothermal nucleic acid amplification using pH-sensitive dyes
    Tanner, Nathan A.
    Zhang, Yinhua
    Evans, Thomas C., Jr.
    [J]. BIOTECHNIQUES, 2015, 58 (02) : 59 - 68
  • [9] Rapid and sensitive detection of Zika virus by reverse transcription loop-mediated isothermal amplification
    Wang, Xuan
    Yin, Fenggui
    Bi, Yuhai
    Cheng, Gong
    Li, Jing
    Hou, Lidan
    Li, Yunlong
    Yang, Baozhi
    Liu, Wenjun
    Yang, Limin
    [J]. JOURNAL OF VIROLOGICAL METHODS, 2016, 238 : 86 - 93
  • [10] Rapid Detection of COVID-19 Coronavirus Using a Reverse Transcriptional Loop-Mediated Isothermal Amplification (RT-LAMP) Diagnostic Platform
    Yu, Lin
    Wu, Shanshan
    Hao, Xiaowen
    Dong, Xue
    Mao, Lingling
    Pelechano, Vicent
    Chen, Wei-Hua
    Yin, Xiushan
    [J]. CLINICAL CHEMISTRY, 2020, 66 (07) : 975 - 977