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

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dc.contributor.authorAli Bektaş
dc.contributor.authorMichael F. Covington
dc.contributor.authorGuy Aidelberg
dc.contributor.authorAnibal Arce
dc.contributor.authorTamara Matute
dc.contributor.authorIsaac Núñez
dc.contributor.authorJulia Walsh
dc.contributor.authorDavid BOUTBOUL
dc.contributor.authorConstance Delaugerre
dc.contributor.authorAriel B. Lindner
dc.contributor.authorFernán Federici
dc.contributor.authorAnitha D. Jayaprakash
dc.date.accessioned2023-05-19T20:45:41Z
dc.date.available2023-05-19T20:45:41Z
dc.date.issued2021
dc.description.abstractThe 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/mu L) 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.
dc.fuente.origenORCID-mayo23
dc.identifier.doi10.3390/v13050742
dc.identifier.urihttps://repositorio.uc.cl/handle/11534/69540
dc.language.isoen
dc.rightsacceso restringido
dc.titleAccessible LAMP-Enabled Rapid Test (ALERT) for Detecting SARS-CoV-2es_ES
dc.typeartículo
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