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Accessible LAMP-Enabled Rapid Test (ALERT) for Detecting SARS-CoV-2

Oakland Genomics Center, 355 30th Street, Oakland, CA 94609, USA
Amaryllis Nucleics, 355 30th Street, Oakland, CA 94609, USA
Center for Research and Interdisciplinarity (CRI), Université de Paris, INSERM U1284, F-75006 Paris, France
Institute for Biological and Medical Engineering, Schools of Engineering, Biology and Medicine, Pontificia Universidad Católica de Chile, Santiago 7820244, Chile
FONDAP Center for Genome Regulation, ANID–Millennium Science Initiative Program–Millennium Institute for Integrative Biology (iBIO), Santiago 8331150, Chile
School of Public Health, University of California Berkeley, Berkeley, CA 94720, USA
Clinical Immunology Department, U976 HIPI, Hôpital Saint Louis, Université de Paris, F-75006 Paris, France
Girihlet Inc., 355 30th Street, Oakland, CA 94609, USA
Authors to whom correspondence should be addressed.
Academic Editors: Bertram Flehmig and Karim Klingel
Viruses 2021, 13(5), 742;
Received: 14 March 2021 / Revised: 8 April 2021 / Accepted: 15 April 2021 / Published: 23 April 2021
(This article belongs to the Special Issue Viral Markers and the Diagnosis of COVID-19)
The coronavirus disease 2019 (COVID-19) pandemic has highlighted bottlenecks in large-scale, frequent testing of populations for infections. Polymerase chain reaction (PCR)-based diagnostic tests are expensive, reliant on centralized labs, can take days to deliver results, and are prone to backlogs and supply shortages. Antigen tests that bind and detect the surface proteins of a virus are rapid and scalable but suffer from high false negative rates. To address this problem, an inexpensive, simple, and robust 60-minute do-it-yourself (DIY) workflow to detect viral RNA from nasal swabs or saliva with high sensitivity (0.1 to 2 viral particles/μL) and specificity (>97% true negative rate) utilizing reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed. ALERT (Accessible LAMP-Enabled Rapid Test) incorporates the following features: (1) increased shelf-life and ambient temperature storage, compared to liquid reaction mixes, by using wax layers to isolate enzymes from other reagents; (2) improved specificity compared to other LAMP end-point reporting methods, by using sequence-specific QUASR (quenching of unincorporated amplification signal reporters); (3) increased sensitivity, compared to methods without purification through use of a magnetic wand to enable pipette-free concentration of sample RNA and cell debris removal; (4) quality control with a nasopharyngeal-specific mRNA target; and (5) co-detection of other respiratory viruses, such as influenza B, by multiplexing QUASR-modified RT-LAMP primer sets. The flexible nature of the ALERT workflow allows easy, at-home and point-of-care testing for individuals and higher-throughput processing for labs and hospitals. With minimal effort, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific primer sets can be swapped out for other targets to repurpose ALERT to detect other viruses, microorganisms, or nucleic acid-based markers. View Full-Text
Keywords: RT-LAMP; point-of-care; biodetection; SARS-CoV-2 RT-LAMP; point-of-care; biodetection; SARS-CoV-2
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MDPI and ACS Style

Bektaş, A.; Covington, M.F.; Aidelberg, G.; Arce, A.; Matute, T.; Núñez, I.; Walsh, J.; Boutboul, D.; Delaugerre, C.; Lindner, A.B.; Federici, F.; Jayaprakash, A.D. Accessible LAMP-Enabled Rapid Test (ALERT) for Detecting SARS-CoV-2. Viruses 2021, 13, 742.

AMA Style

Bektaş A, Covington MF, Aidelberg G, Arce A, Matute T, Núñez I, Walsh J, Boutboul D, Delaugerre C, Lindner AB, Federici F, Jayaprakash AD. Accessible LAMP-Enabled Rapid Test (ALERT) for Detecting SARS-CoV-2. Viruses. 2021; 13(5):742.

Chicago/Turabian Style

Bektaş, Ali, Michael F. Covington, Guy Aidelberg, Anibal Arce, Tamara Matute, Isaac Núñez, Julia Walsh, David Boutboul, Constance Delaugerre, Ariel B. Lindner, Fernán Federici, and Anitha D. Jayaprakash. 2021. "Accessible LAMP-Enabled Rapid Test (ALERT) for Detecting SARS-CoV-2" Viruses 13, no. 5: 742.

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