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Diagnostics (Basel) . A S imple, A ffordable, R apid, S tabilized, Co lorimetric, V ersatile RT-LAMP Assay to Detect SARS-CoV-2

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  • Diagnostics (Basel) . A S imple, A ffordable, R apid, S tabilized, Co lorimetric, V ersatile RT-LAMP Assay to Detect SARS-CoV-2


    Diagnostics (Basel)


    . 2021 Mar 4;11(3):438.
    doi: 10.3390/diagnostics11030438.
    A S imple, A ffordable, R apid, S tabilized, Co lorimetric, V ersatile RT-LAMP Assay to Detect SARS-CoV-2


    Juan García-Bernalt Diego 1 , Pedro Fernández-Soto 1 , Marta Domínguez-Gil 2 , Moncef Belhassen-García 1 3 , Juan Luis Muñoz Bellido 4 , Antonio Muro 1



    Affiliations

    Abstract

    The SARS-CoV-2 pandemic has forced all countries worldwide to rapidly develop and implement widespread testing to control and manage the Coronavirus Disease 2019 (COVID-19). reverse-transcription (RT)-qPCR is the gold standard molecular diagnostic method for COVID-19, mostly in automated testing platforms. These systems are accurate and effective, but also costly, time-consuming, high-technological, infrastructure-dependent, and currently suffer from commercial reagent supply shortages. The reverse-transcription loop-mediated isothermal amplification (RT-LAMP) can be used as an alternative testing method. Here, we present a novel versatile (real-time and colorimetric) RT-LAMP for the simple (one-step), affordable (~1.7 €/sample), and rapid detection of SARS-CoV-2 targeting both ORF1ab and N genes of the novel virus genome. We demonstrate the assay on RT-qPCR-positive clinical samples, obtaining most positive results under 25 min. In addition, a novel 30-min one-step drying protocol has been developed to stabilize the RT-LAMP reaction mixtures, allowing them to be stored at room temperature functionally for up to two months, as predicted by the Q10. This Dry-RT-LAMP methodology is suitable for potentially ready-to-use COVID-19 diagnosis. After further testing and validation, it could be easily applied both in developed and in low-income countries yielding rapid and reliable results.

    Keywords: RT-LAMP; SARS-CoV-2; dry-RT-LAMP; molecular diagnostics; point-of-care.

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