tetano
Editor, Senior Moderator
Front Mol Biosci
. 2022 Apr 1;9:801309.
doi: 10.3389/fmolb.2022.801309. eCollection 2022.
A Rapid, Highly Sensitive and Open-Access SARS-CoV-2 Detection Assay for Laboratory and Home Testing
Max J Kellner[SUP] 1 2 3 4 [/SUP], James J Ross[SUP] 2 3 [/SUP], Jakob Schnabl[SUP] 2 3 [/SUP], Marcus P S Dekens[SUP] 1 [/SUP], Martin Matl[SUP] 1 2 [/SUP], Robert Heinen[SUP] 1 2 [/SUP], Irina Grishkovskaya[SUP] 1 [/SUP], Benedikt Bauer[SUP] 1 [/SUP], Johannes Stadlmann[SUP] 1 2 5 [/SUP], Luis Menéndez-Arias[SUP] 6 [/SUP], Andrew D Straw[SUP] 7 [/SUP], Robert Fritsche-Polanz[SUP] 8 [/SUP], Marianna Traugott[SUP] 9 [/SUP], Tamara Seitz[SUP] 9 [/SUP], Alexander Zoufaly[SUP] 9 [/SUP], Manuela Födinger[SUP] 8 10 [/SUP], Christoph Wenisch[SUP] 9 [/SUP], Johannes Zuber[SUP] 1 11 [/SUP], Andrea Pauli[SUP] 1 [/SUP], Julius Brennecke[SUP] 2 [/SUP]
Affiliations
Abstract
RT-qPCR-based diagnostic tests play important roles in combating virus-caused pandemics such as Covid-19. However, their dependence on sophisticated equipment and the associated costs often limits their widespread use. Loop-mediated isothermal amplification after reverse transcription (RT-LAMP) is an alternative nucleic acid detection method that overcomes these limitations. Here, we present a rapid, robust, and sensitive RT-LAMP-based SARS-CoV-2 detection assay. Our 40-min procedure bypasses the RNA isolation step, is insensitive to carryover contamination, and uses a colorimetric readout that enables robust SARS-CoV-2 detection from various sample types. Based on this assay, we have increased sensitivity and scalability by adding a nucleic acid enrichment step (Bead-LAMP), developed a version for home testing (HomeDip-LAMP), and identified open-source RT-LAMP enzymes that can be produced in any molecular biology laboratory. On a dedicated website, rtlamp.org (DOI: 10.5281/zenodo.6033689), we provide detailed protocols and videos. Our optimized, general-purpose RT-LAMP assay is an important step toward population-scale SARS-CoV-2 testing.
Keywords: COVID-19 diagnostics; RT-LAMP; SARS-CoV-2; coronavirus; isothermal amplification; open-source.
. 2022 Apr 1;9:801309.
doi: 10.3389/fmolb.2022.801309. eCollection 2022.
A Rapid, Highly Sensitive and Open-Access SARS-CoV-2 Detection Assay for Laboratory and Home Testing
Max J Kellner[SUP] 1 2 3 4 [/SUP], James J Ross[SUP] 2 3 [/SUP], Jakob Schnabl[SUP] 2 3 [/SUP], Marcus P S Dekens[SUP] 1 [/SUP], Martin Matl[SUP] 1 2 [/SUP], Robert Heinen[SUP] 1 2 [/SUP], Irina Grishkovskaya[SUP] 1 [/SUP], Benedikt Bauer[SUP] 1 [/SUP], Johannes Stadlmann[SUP] 1 2 5 [/SUP], Luis Menéndez-Arias[SUP] 6 [/SUP], Andrew D Straw[SUP] 7 [/SUP], Robert Fritsche-Polanz[SUP] 8 [/SUP], Marianna Traugott[SUP] 9 [/SUP], Tamara Seitz[SUP] 9 [/SUP], Alexander Zoufaly[SUP] 9 [/SUP], Manuela Födinger[SUP] 8 10 [/SUP], Christoph Wenisch[SUP] 9 [/SUP], Johannes Zuber[SUP] 1 11 [/SUP], Andrea Pauli[SUP] 1 [/SUP], Julius Brennecke[SUP] 2 [/SUP]
Affiliations
- PMID: 35433827
- PMCID: PMC9011764
- DOI: 10.3389/fmolb.2022.801309
Abstract
RT-qPCR-based diagnostic tests play important roles in combating virus-caused pandemics such as Covid-19. However, their dependence on sophisticated equipment and the associated costs often limits their widespread use. Loop-mediated isothermal amplification after reverse transcription (RT-LAMP) is an alternative nucleic acid detection method that overcomes these limitations. Here, we present a rapid, robust, and sensitive RT-LAMP-based SARS-CoV-2 detection assay. Our 40-min procedure bypasses the RNA isolation step, is insensitive to carryover contamination, and uses a colorimetric readout that enables robust SARS-CoV-2 detection from various sample types. Based on this assay, we have increased sensitivity and scalability by adding a nucleic acid enrichment step (Bead-LAMP), developed a version for home testing (HomeDip-LAMP), and identified open-source RT-LAMP enzymes that can be produced in any molecular biology laboratory. On a dedicated website, rtlamp.org (DOI: 10.5281/zenodo.6033689), we provide detailed protocols and videos. Our optimized, general-purpose RT-LAMP assay is an important step toward population-scale SARS-CoV-2 testing.
Keywords: COVID-19 diagnostics; RT-LAMP; SARS-CoV-2; coronavirus; isothermal amplification; open-source.