Publications
Sensors & diagnosticsMar 2024 |
3
(
3
),
421-430
DOI:
10.1039/d3sd00246b

SARS-CoV-2 recombinase polymerase amplification assay with lateral flow readout and duplexed full process internal control

Martin, Coleman D; Bender, Andrew T; Sullivan, Benjamin P; Lillis, Lorraine; Boyle, David S; Posner, Jonathan D
Product Used
NGS
Abstract
Nucleic acid amplification tests for the detection of SARS-CoV-2 have been an important testing mechanism for the COVID-19 pandemic. While these traditional nucleic acid diagnostic methods are highly sensitive and selective, they are not suited to home or clinic-based uses. Comparatively, rapid antigen tests are cost-effective and user friendly but lack in sensitivity and specificity. Here we report on the development of a one-pot, duplexed reverse transcriptase recombinase polymerase amplification SARS-CoV-2 assay with MS2 bacteriophage as a full process control. Detection is carried out with either real-time fluorescence or lateral flow readout with an analytical sensitivity of 50 copies per reaction. Unlike previously published assays, the RNA-based MS2 bacteriophage control reports on successful operation of lysis, reverse transcription, and amplification. This SARS-CoV-2 assay features highly sensitive detection, visual readout through an LFA strip, results in less than 25 minutes, minimal instrumentation, and a useful process internal control to rule out false negative test results.This journal is
Product Used
NGS

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