Rapid isothermal amplification and portable detection system for SARS-CoV-2
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Abstract
An important limitation of current assays for the detection of SARS-CoV-2 stems from their reliance on time-consuming, labor-intensive, and laboratory-based protocols for viral isolation, lysis, and removal of inhibiting materials. While RT-PCR remains the gold standard for performing clinical diagnostics to amplify the RNA sequences, there is an urgent need for alternative testing platforms that are rapid, accurate, simple, and portable. Here, we demonstrate isothermal RT-LAMP nucleic acid-based detection of SARS-CoV-2 with an additively manufactured cartridge and a smartphone-based instrument for testing that can be performed at the point of sample collection.
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SciScore for 10.1101/2020.05.21.108381: (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 First, thermally lysed patient sample and RT-LAMP reagents are injected through the female luer lock connectors from two separate syringes without the use of microfluidic pumps. Table 2: Resources
Software and Algorithms Sentences Resources Finally, 0.47U/μL BST 2.0 WarmStart DNA Polymerase (New England Bioloabs), 0.3 U/μL New England Bioloabssuggested: NoneOff Chip amplification data analysis: The off-chip RT-LAMP fluorescence curves and amplification threshold bar graphs were analyzed using a MATLAB script and plotted using GraphPad Prism 7. MATLABsuggest…SciScore for 10.1101/2020.05.21.108381: (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 First, thermally lysed patient sample and RT-LAMP reagents are injected through the female luer lock connectors from two separate syringes without the use of microfluidic pumps. Table 2: Resources
Software and Algorithms Sentences Resources Finally, 0.47U/μL BST 2.0 WarmStart DNA Polymerase (New England Bioloabs), 0.3 U/μL New England Bioloabssuggested: NoneOff Chip amplification data analysis: The off-chip RT-LAMP fluorescence curves and amplification threshold bar graphs were analyzed using a MATLAB script and plotted using GraphPad Prism 7. MATLABsuggested: (MATLAB, RRID:SCR_001622)GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)Chip Image and data analysis: Fluorescence images were recorded with IP Webcam in smartphones and were saved in JPG format from which fluorescence intensity and baseline fluorescence was analyzed on Image J. Image Jsuggested: (ImageJ, RRID:SCR_003070)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 found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- 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.
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