By contrast, repeated testing of large groups of individuals, regardless of symptoms or contact status, is predicted to be an effective measure to decrease SARS-CoV-2 transmission 3, 4, 5. Current containment strategies based on ‘test–trace–isolate’ face major issues: (1) many infected individuals do not show any symptoms and, therefore, remain untested 2 (2) supply chain issues limit testing capacity and (3) the successive (rather than parallel) testing of contact individuals causes a substantial lag in identifying infection chains, resulting in undetected spread due to delayed diagnosis. With a time-to-result of fewer than 24 h, low cost and little new infrastructure requirement, LAMP-Seq can be readily deployed for frequent testing as part of an integrated public health surveillance program.Īs of February 2021, the global spread of a novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has resulted in over 109 million confirmed cases, and approximately 2.4 million deaths have been attributed to Coronavirus Disease 2019 (COVID-19) 1. Clinical validation of an open-source protocol with 676 swab samples, 98 of which were deemed positive by standard RT–qPCR, demonstrated 100% sensitivity in individuals with cycle threshold values of up to 33 and a specificity of 99.7%, at a very low material cost. Using commercial reagents, LAMP-Seq has a limit of detection of ~2.2 molecules per µl at 95% confidence and near-perfect specificity for severe acute respiratory syndrome coronavirus 2 given its sequence readout.
Unpurified biosamples are barcoded and amplified in a single heat step, and pooled products are analyzed en masse by sequencing. Here we present LAMP-Seq, a modified, highly scalable reverse transcription loop-mediated isothermal amplification (RT–LAMP) method. Nature Biotechnology volume 39, pages 1556–1562 ( 2021) Cite this articleįrequent testing of large population groups combined with contact tracing and isolation measures will be crucial for containing Coronavirus Disease 2019 outbreaks. (1.LAMP-Seq enables sensitive, multiplexed COVID-19 diagnostics using molecular barcoding
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