tetano
Editor, Senior Moderator
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 Garc?a-Bernalt Diego[SUP] 1 [/SUP], Pedro Fern?ndez-Soto[SUP] 1 [/SUP], Marta Dom?nguez-Gil[SUP] 2 [/SUP], Moncef Belhassen-Garc?a[SUP] 1 3 [/SUP], Juan Luis Mu?oz Bellido[SUP] 4 [/SUP], Antonio Muro[SUP] 1 [/SUP]
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 Q[SUB]10[/SUB]. 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.
. 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 Garc?a-Bernalt Diego[SUP] 1 [/SUP], Pedro Fern?ndez-Soto[SUP] 1 [/SUP], Marta Dom?nguez-Gil[SUP] 2 [/SUP], Moncef Belhassen-Garc?a[SUP] 1 3 [/SUP], Juan Luis Mu?oz Bellido[SUP] 4 [/SUP], Antonio Muro[SUP] 1 [/SUP]
Affiliations
- PMID: 33806456
- DOI: 10.3390/diagnostics11030438
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 Q[SUB]10[/SUB]. 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.