Development of Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) Assays Targeting SARS-CoV-2
This article has been Reviewed by the following groups
Discuss this preprint
Start a discussion What are Sciety discussions?Listed in
- Evaluated articles (ScreenIT)
Abstract
Epidemics of Coronavirus Disease 2019 (COVID-19) now have more than 100,000 confirmed cases worldwide. Diagnosis of COVID-19 is currently performed by RT-qPCR methods, but the capacity of RT-qPCR methods is limited by its requirement of high-level facilities and instruments. Here, we developed and evaluated RT-LAMP assays to detect genomic RNA of SARS-CoV-2, the causative virus of COVID-19. RT-LAMP assays in this study can detect as low as 100 copies of SARS-CoV-2 RNA. Cross-reactivity of RT-LAMP assays to other human Coronaviruses was not observed. We also adapted a colorimetric detection method for our RT-LAMP assay so that the tests potentially performed in higher throughput.
Article activity feed
-
SciScore for 10.1101/2020.03.09.983064: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Experimental Models: Cell Lines Sentences Resources hCoV-229E and hCoV-OC43 viral RNA were isolated from culture media of infected MRC-5 cells (ATCC® CCL-171™). MRC-5suggested: NoneMERS-CoV RNA was isolated from cell pellet lysate of infected Vero cells (ATCC® CCL-81™). Verosuggested: NoneSoftware and Algorithms Sentences Resources LAMP and RT-LAMP reaction: LAMP reaction was performed with reaction mixture containing following components: 1.6 μM FIP/BIP primers, 0.2 μM F3/B3 primers, 0.4 … SciScore for 10.1101/2020.03.09.983064: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Experimental Models: Cell Lines Sentences Resources hCoV-229E and hCoV-OC43 viral RNA were isolated from culture media of infected MRC-5 cells (ATCC® CCL-171™). MRC-5suggested: NoneMERS-CoV RNA was isolated from cell pellet lysate of infected Vero cells (ATCC® CCL-81™). Verosuggested: NoneSoftware and Algorithms Sentences Resources LAMP and RT-LAMP reaction: LAMP reaction was performed with reaction mixture containing following components: 1.6 μM FIP/BIP primers, 0.2 μM F3/B3 primers, 0.4 μM LF/LB primers, 1x Isothermal Amplification Buffer II (NEB, 20 mM Tris-HCl pH 8.8, 10 mM (NH4)2SO4, 150 mM KCl, 2 mM MgSO4, 0.1% Tween® 20), 6 mM MgSO4 (NEB, final 8mM Mg2+), 1.4 mM each dNTP (Enzynomics), 0.4 μM SYTO-9 (Invitrogen) and 6 U Bst3.0 DNA polymerase (NEB) in total 15 μl reaction volume. LAMPsuggested: (LAMP, RRID:SCR_001740)Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- No conflict of interest statement was detected. If there are no conflicts, we encourage authors to explicit state so.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
-
