Rapid Isothermal Amplification and Portable Detection System for SARS-CoV-2
Anurup Ganguli,Ariana Mostafa,Jacob Berger,Mehmet Y. Aydin,Fu Sun,Enrique Valera,Brian T. Cunningham,William P. King,Rashid Bashir +8 more
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TLDR
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 are provided, including the demonstration of a smartphone-based point-of-care device that can be used at the point of sample collection.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 of SARS-CoV-2 virus in less than 40 minutes using an additively manufactured cartridge and a smartphone-based reader. Finally, we explore the speed and cost advantages by comparing the required resources and workflows with RT-PCR. This work could accelerate the development and availability of SARS-CoV-2 diagnostics by proving alternatives to conventional laboratory benchtop tests.read more
Citations
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Rapid, point-of-care antigen and molecular-based tests for diagnosis of SARS-CoV-2 infection.
Jacqueline Dinnes,Jacqueline Dinnes,Jonathan J Deeks,Ada Adriano,Sarah Berhane,Clare Davenport,Sabine Dittrich,Devy Emperador,Yemisi Takwoingi,Jane Cunningham,Sophie Beese,Janine Dretzke,Lavinia Ferrante di Ruffano,Isobel Marion Harris,Malcolm J Price,Sian Taylor-Phillips,Lotty Hooft,Mariska M.G. Leeflang,René Spijker,Ann Van den Bruel +19 more
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Saliva-Based Molecular Testing for SARS-CoV-2 that Bypasses RNA Extraction
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