LAMP-Seq enables sensitive, multiplexed COVID-19 diagnostics using molecular barcoding.
Nat Biotechnol
; 39(12): 1556-1562, 2021 12.
Article
en En
| MEDLINE
| ID: mdl-34188222
ABSTRACT
Frequent testing of large population groups combined with contact tracing and isolation measures will be crucial for containing Coronavirus Disease 2019 outbreaks. Here we present LAMP-Seq, a modified, highly scalable reverse transcription loop-mediated isothermal amplification (RT-LAMP) method. Unpurified biosamples are barcoded and amplified in a single heat step, and pooled products are analyzed en masse by sequencing. 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. 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. 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.
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Técnicas de Amplificación de Ácido Nucleico
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Técnicas de Diagnóstico Molecular
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Prueba de COVID-19
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COVID-19
Tipo de estudio:
Diagnostic_studies
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Guideline
Límite:
Humans
Idioma:
En
Año:
2021
Tipo del documento:
Article