Sep 08, 2020
XPRIZE SHINE - Paper-based SARS-CoV-2 Saliva Test
- Jon Arizti-Sanz1,2,
- Catherine A. Freije1,
- Chloe K. Boehm1,
- Sameed M. Siddiqui1,3,
- Allen M. Goodman1,
- Tinna-Solveig F. Kosoko-Thoroddsen1,
- A'Doriann Y. Bradley1,
- Jeremy Johnson1,
- Pardis C. Sabeti1,4,5,6,7,
- Cameron Myhrvold1,8,7
- 1Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA.;
- 2Harvard-MIT Program in Health Sciences and Technology, 77 Massachusetts Ave., Cambridge, MA 02139, USA.;
- 3Computational and Systems Biology PhD Program, MIT, Cambridge, MA 02139, USA.;
- 4Harvard T.H. Chan School of Public Health, 677 Huntington Avenue, Boston, MA 02115, USA.;
- 5Department of Organismic and Evolutionary Biology, Harvard University, 26 Oxford Street, Cambridge, MA 02138, USA.;
- 6Howard Hughes Medical Institute, Chevy Chase, MD 20815, USA.;
- 7Massachusetts Consortium on Pathogen Readiness, Boston, MA, USA.;
- 8Department of Organismic and Evolutionary Biology, Harvard University, 26 Oxford Street, Cambridge, MA 02138, USA
- XPRIZE Rapid Covid Testing
Protocol Citation: Jon Arizti-Sanz, Catherine A. Freije, Chloe K. Boehm, Sameed M. Siddiqui, Allen M. Goodman, Tinna-Solveig F. Kosoko-Thoroddsen, A'Doriann Y. Bradley, Jeremy Johnson, Pardis C. Sabeti, Cameron Myhrvold 2020. XPRIZE SHINE - Paper-based SARS-CoV-2 Saliva Test. protocols.io https://dx.doi.org/10.17504/protocols.io.bk2fkybn
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: September 08, 2020
Last Modified: September 08, 2020
Protocol Integer ID: 41767
Keywords: CRISPR, SARS-CoV-2, nucleic acid diagnostic,
Disclaimer
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Abstract
This protocol describes how to perform a SHINE paper-based assay to detect SARS-CoV-2 RNA from a self-collected saliva sample. This protocol is intended for in-home use. All enzymatic components are provided as a single-test freeze-dried pellet for shelf-stable storage, and all steps of the protocol are performed at ambient temperature. The protocol presented here is an improved version of the method presented in Arizti-Sanz J*, Freije CA*, et al. Integrated sample inactivation, amplification, and Cas13-based detection of SARS-CoV-2. bioRxiv (2020).
Image Attribution
biorender.com
Materials
MATERIALS
Nalgene™ Dropper Bottles with Control Dispensing Tip, 4mL, whiteThermo FisherCatalog #2750-9125
Screw cap tube 5 mL sterileThomas ScientificCatalog #1188R46
FastAmp® Viral and Cell Solution for Covid-19 Testing Solution BCatalog #4633
Reagent Mix A (Paper-based SARS-CoV-2 resuspension mix)
Lyophilized Reagent Mix B (Paper-based SARS-CoV-2 detection mix)
HybriDetect — Universal Lateral Flow Assay KitCatalog #MGHD 1
Inoculating Loops and Needles Sterile 10 uLVWR InternationalCatalog #12000-810
STEP MATERIALS
FastAmp® Viral and Cell Solution for Covid-19 Testing Solution BCatalog #4633
Screw cap tube 5 mL sterileThomas ScientificCatalog #1188R46
Lyophilized Reagent Mix B (Paper-based SARS-CoV-2 detection mix)
Nalgene™ Dropper Bottles, 15mL, whiteThermo FisherCatalog #2751-9050
Reagent Mix A (Paper-based SARS-CoV-2 resuspension mix)
Inoculating Loops and Needles Sterile 10 uLVWR InternationalCatalog #12000-810
HybriDetect — Universal Lateral Flow Assay KitCatalog #MGHD 1
The necessary volume of FastAmp Viral and Cell Solution is provided in the screw cap tube used for sample collection. All enzymatic components, reagents, and compatible buffers required for SARS-CoV-2 detection are included within Lyophilized Reagent Mix B and are reconstituted with Reagent Mix A (see protocol for details). A smartphone or smart device is necessary for automated interpretation of the SARS-CoV-2 detection results using the HandLens application. The required volume of Reagent Mix A is provided in the Nalgene Dropper Bottle.
Protocol materials
Nalgene™ Dropper Bottles, 15mL, whiteThermo FisherCatalog #2751-9050
HybriDetect — Universal Lateral Flow Assay KitCatalog #MGHD 1
Inoculating Loops and Needles Sterile 10 uLVWR International (Avantor)Catalog #12000-810
FastAmp® Viral and Cell Solution for Covid-19 Testing Solution BCatalog #4633
Reagent Mix A (Paper-based SARS-CoV-2 resuspension mix)
FastAmp® Viral and Cell Solution for Covid-19 Testing Solution BCatalog #4633
Lyophilized Reagent Mix B (Paper-based SARS-CoV-2 detection mix)
Lyophilized Reagent Mix B (Paper-based SARS-CoV-2 detection mix)
Screw cap tube 5 mL sterileThomas ScientificCatalog #1188R46
Screw cap tube 5 mL sterileThomas ScientificCatalog #1188R46
Nalgene™ Dropper Bottles with Control Dispensing Tip, 4mL, whiteThermo FisherCatalog #2750-9125
Inoculating Loops and Needles Sterile 10 uLVWR International (Avantor)Catalog #12000-810
HybriDetect — Universal Lateral Flow Assay KitCatalog #MGHD 1
Reagent Mix A (Paper-based SARS-CoV-2 resuspension mix)
FastAmp® Viral and Cell Solution for Covid-19 Testing Solution BCatalog #4633
Screw cap tube 5 mL sterileThomas ScientificCatalog #1188R46
Nalgene™ Dropper Bottles, 15mL, whiteThermo FisherCatalog #2751-9050
Reagent Mix A (Paper-based SARS-CoV-2 resuspension mix)
Lyophilized Reagent Mix B (Paper-based SARS-CoV-2 detection mix)
Inoculating Loops and Needles Sterile 10 uLVWR International (Avantor)Catalog #12000-810
HybriDetect — Universal Lateral Flow Assay KitCatalog #MGHD 1
Safety warnings
Please take care with potentially infectious sample material that does not come into contact with the provided viral lysis solution contained within the saliva sample collection tube.
Before start
Download the HandLens application on the user-provided smart device (smartphone, tablet, etc.). Wash hands prior to starting the protocol.
Sample Collection and Viral Lysis
Sample Collection and Viral Lysis
Expel approximately one drop of saliva into the sample collection tube and cap the tube.
Saliva collection tube contains necessary volume of FastAmp® Viral and Cell Solution.
FastAmp® Viral and Cell Solution for Covid-19 Testing Solution BCatalog #4633
Screw cap tube 5 mL sterileThomas ScientificCatalog #1188R46
Mix saliva sample and FastAmp® Viral and Cell Solution by shaking the closed sample collection tube for 00:00:10
Wait 00:05:00 incubating sample at Room temperature before proceeding to Step 4.
SARS-CoV-2 Detection
SARS-CoV-2 Detection
Add the entire volume in the dropper bottle containing Reagent Mix A to the tube containing lyophilized Reagent Mix B. Cap the tube.
Nalgene™ Dropper Bottles, 15mL, whiteThermo FisherCatalog #2751-9050
Reagent Mix A (Paper-based SARS-CoV-2 resuspension mix)
Lyophilized Reagent Mix B (Paper-based SARS-CoV-2 detection mix)
Mix Reagent Mix A and B by shaking for approximately 00:00:10 .
Dip the inoculation loop into the sample collection tube (a small layer of liquid should be contained within the loop).
Inoculating Loops and Needles Sterile 10 uLVWR International (Avantor)Catalog #12000-810
Transfer liquid in inoculation loop to the Reagent Mix A and B tube by dipping the loop into tube and stirring for 00:00:05 .. Remove and discard the inoculation loop and cap the tube.
Mix the sample combined with Reagent Mix A and B by shaking for approximately 00:00:10 .
Wait 01:30:00 , incubating sample at Room temperature , before proceeding to Step 10.
Paper-based Readout and Automated Analysis
Paper-based Readout and Automated Analysis
Open the Sample-Reagent Mix A and B tube and place the test strip into the liquid with the arrows on the test strip pointing upward and towards you.
HybriDetect — Universal Lateral Flow Assay KitCatalog #MGHD 1
Wait 00:05:00 with sample at Room temperature for visible horizontal bands to appear on the test strip.
With the user-provided smart device such as a smartphone, open the HandLens application and select paper-based as the test type.
Take a photo of the test strip, and select upload. The result of the test will appear on the smart device screen.