@article{6a29ec3619434be19abd7dfcfe45cd2e,
title = "Rapid isothermal amplification and portable detection system for SARS-CoV-2",
abstract = "The COVID-19 pandemic provides an urgent example where a gap exists between availability of state-of-the-art diagnostics and current needs. As assay protocols and primer sequences become widely known, many laboratories perform diagnostic tests using methods such as RT-PCR or reverse transcription loop mediated isothermal amplification (RT-LAMP). Here, we report an RT-LAMP isothermal assay for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus and demonstrate the assay on clinical samples using a simple and accessible point-of-care (POC) instrument. We characterized the assay by dipping swabs into synthetic nasal fluid spiked with the virus, moving the swab to viral transport medium (VTM), and sampling a volume of the VTM to perform the RT-LAMP assay without an RNA extraction kit. The assay has a limit of detection (LOD) of 50 RNA copies per μL in the VTM solution within 30 min. We further demonstrate our assay by detecting SARS-CoV-2 viruses from 20 clinical samples. Finally, we demonstrate a portable and real-time POC device to detect SARS-CoV-2 from VTM samples using an additively manufactured three-dimensional cartridge and a smartphone-based reader. The POC system was tested using 10 clinical samples, and was able to detect SARS-CoV-2 from these clinical samples by distinguishing positive samples from negative samples after 30 min. The POC tests are in complete agreement with RT-PCR controls. This work demonstrates an alternative pathway for SARS-CoV-2 diagnostics that does not require conventional laboratory infrastructure, in settings where diagnosis is required at the point of sample collection.",
keywords = "COVID-19 diagnostics, Point-of-care, RT-LAMP, SARS-CoV-2, Smartphone reader",
author = "Anurup Ganguli and Ariana Mostafa and Jacob Berger and Aydin, {Mehmet Y.} and Fu Sun and {Stewart de Ramirez}, {Sarah A.} and Enrique Valera and Cunningham, {Brian T.} and King, {William P.} and Rashid Bashir",
note = "Funding Information: Inactivated, NR-52286. We thank the staff at the Holonyak Micro and Nanotechnology Laboratory at University of Illinois at Urbana–Champaign (UIUC) for facilitating the research and the funding from University of Illinois. Microfluidic diagnostic cartridges were provided by Fast Radius Inc. R.B. and E.V. acknowledge support for A.G. from NIH R21 AI146865A. We acknowledge partial support of A.M. by a cooperative agreement with Purdue University and the Agricultural Research Service of the United States Department of Agriculture (via sub-award 8000074077 to UIUC). We acknowledge the support of Cooperative Agreement D19AC00012 awarded by the Defense Funding Information: Advanced Research Projects Agency of the US Department of Defense to W.P.K. and R.B. to support M.Y.A., J.B., and E.V. NSF support of F.S. under Grant 1534126 is acknowledged. We also thank Sara Riggenbach, Gabriel Koch, and Bill Bond of OSH Healthcare, Peoria, IL, for their support of the IRB-1602513 and patient sample acquisition for this study. We acknowledge the support of NSF Rapid Response Research (RAPID) grant (Award 2028431) and Jump Applied Research through Community Health through Engineering and Simulation (ARCHES) endowment through the Health Care Engineering Systems Center at UIUC. Funding Information: The following reagents were deposited by the Centers for Disease Control and Prevention and obtained through BEI Resources, National Institute of Allergy and Infectious Diseases (NIAID), NIH: 1) Genomic RNA from SARS-Related Coronavirus 2, Isolate USA-WA1/ 2020, NR-52285; 2) SARS-Related Coronavirus 2, Isolate USA-WA1/2020, Heat Inactivated, NR-52286. We thank the staff at the Holonyak Micro and Nanotechnology Laboratory at University of Illinois at Urbana?Champaign (UIUC) for facilitating the research and the funding from University of Illinois. Microfluidic diagnostic cartridges were provided by Fast Radius Inc. R.B. and E.V. acknowledge support for A.G. from NIH R21 AI146865A. We acknowledge partial support of A.M. by a cooperative agreement with Purdue University and the Agricultural Research Service of the United States Department of Agriculture (via sub-award 8000074077 to UIUC). We acknowledge the support of Cooperative Agreement D19AC00012 awarded by the Defense Advanced Research Projects Agency of the US Department of Defense to W.P.K. and R.B. to support M.Y.A., J.B., and E.V. NSF support of F.S. under Grant 1534126 is acknowledged. We also thank Sara Riggenbach, Gabriel Koch, and Bill Bond of OSH Healthcare, Peoria, IL, for their support of the IRB-1602513 and patient sample acquisition for this study. We acknowledge the support of NSF Rapid Response Research (RAPID) grant (Award 2028431) and Jump Applied Research through Community Health through Engineering and Simulation (ARCHES) endowment through the Health Care Engineering Systems Center at UIUC. Publisher Copyright: {\textcopyright} 2020 National Academy of Sciences. All rights reserved.",
year = "2020",
month = sep,
day = "15",
doi = "10.1073/pnas.2014739117",
language = "English (US)",
volume = "117",
pages = "22727--22735",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
publisher = "National Academy of Sciences",
number = "37",
}