Rapid Isothermal Amplification and Portable Detection System for SARS-CoV-2

BioRxiv : the Preprint Server for Biology
Anurup GanguliRashid Bashir

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 details and primer sequences become widely known, many laboratories could perform diagnostic tests using methods such as RT-PCR or isothermal RT-LAMP amplification. A key advantage of RT-LAMP based approaches compared to RT-PCR is that RT-LAMP is known to be robust in detecting targets from unprocessed samples. In addition, RT-LAMP assays are performed at a constant temperature enabling speed, simplicity, and point-of-use testing. Here, we provide the details of an RT-LAMP isothermal assay for the detection of SARS-CoV-2 virus with performance comparable to currently approved tests using RT-PCR. We characterize the assay by introducing swabs in virus spiked synthetic nasal fluids, moving the swab to viral transport medium (VTM), and using a volume of that VTM for performing the amplification without an RNA extraction kit. The assay has a Limit-of-Detection (LOD) of 50 RNA copies/μL in the VTM solution within 20 minutes, and LOD of 5000 RNA copies/μL in the nasal solution. Additionally, we show the utility of this assay for real-time point-of-use testing by demonstrating detection o...Continue Reading

Citations

Aug 28, 2020·The Cochrane Database of Systematic Reviews·Jacqueline DinnesUNKNOWN Cochrane COVID-19 Diagnostic Test Accuracy Group
Jan 25, 2021·Biosensors & Bioelectronics·Hwang-Soo KimSehyun Shin
Mar 25, 2021·The Cochrane Database of Systematic Reviews·Jacqueline DinnesUNKNOWN Cochrane COVID-19 Diagnostic Test Accuracy Group
May 14, 2021·Clinical Microbiology Reviews·Seyed Hamid Safiabadi TaliSana Jahanshahi-Anbuhi
Sep 22, 2021·ACS Synthetic Biology·Ahmed MahasMagdy M Mahfouz

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Methods Mentioned

BETA
PCR
chip

Software Mentioned

BLASTn
LAMP
GeneXpert
Primerexplorerv4
Image J

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