tetano
Editor, Senior Moderator
Bio Protoc
. 2021 Mar 20;11(6):e3964.
doi: 10.21769/BioProtoc.3964.
Colorimetric RT-LAMP and LAMP-sequencing forDetecting SARS-CoV-2 RNA in Clinical Samples
Konrad Herbst[SUP] 1 [/SUP], Matthias Meurer[SUP] 1 [/SUP], Daniel Kirrmaier[SUP] 2 [/SUP], Simon Anders[SUP] 1 [/SUP], Michael Knop[SUP] 1 2 3 [/SUP], Viet Loan Dao Thi[SUP] 4 [/SUP]
Affiliations
Abstract
During pandemics, such as the one caused by SARS-CoV-2 coronavirus, simple methods to rapidly test large numbers of people are needed. As a faster and less resource-demanding alternative to detect viral RNA by conventional qPCR, we used reverse transcription loop-mediated isothermal amplification (RT-LAMP). We previously established colorimetric RT-LAMP assays on both purified and unpurified SARS-CoV-2 clinical specimens and further developed a multiplexed sequencing protocol (LAMP-sequencing) to analyze the outcome of many RT-LAMP reactions at the same time (Dao Thi et al., 2020). Extending on this work, we hereby provide step-by-step protocols for both RT-LAMP assays and read-outs.
Keywords: LAMP-sequencing; RT-LAMP; SARS-CoV-2 detection; Tn5 tagmentation; colorimetric assay.
. 2021 Mar 20;11(6):e3964.
doi: 10.21769/BioProtoc.3964.
Colorimetric RT-LAMP and LAMP-sequencing forDetecting SARS-CoV-2 RNA in Clinical Samples
Konrad Herbst[SUP] 1 [/SUP], Matthias Meurer[SUP] 1 [/SUP], Daniel Kirrmaier[SUP] 2 [/SUP], Simon Anders[SUP] 1 [/SUP], Michael Knop[SUP] 1 2 3 [/SUP], Viet Loan Dao Thi[SUP] 4 [/SUP]
Affiliations
- PMID: 33855122
- PMCID: PMC8032487 (available on 2022-03-20)
- DOI: 10.21769/BioProtoc.3964
Abstract
During pandemics, such as the one caused by SARS-CoV-2 coronavirus, simple methods to rapidly test large numbers of people are needed. As a faster and less resource-demanding alternative to detect viral RNA by conventional qPCR, we used reverse transcription loop-mediated isothermal amplification (RT-LAMP). We previously established colorimetric RT-LAMP assays on both purified and unpurified SARS-CoV-2 clinical specimens and further developed a multiplexed sequencing protocol (LAMP-sequencing) to analyze the outcome of many RT-LAMP reactions at the same time (Dao Thi et al., 2020). Extending on this work, we hereby provide step-by-step protocols for both RT-LAMP assays and read-outs.
Keywords: LAMP-sequencing; RT-LAMP; SARS-CoV-2 detection; Tn5 tagmentation; colorimetric assay.