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Authors: Shripal Gandhi, Santo Purnama, Keyur Patel, Praveen Sukumara, Dr Rinu R Ravi
Group: Coronavirus Method Development Community, SensingSelfMission, XPRIZE Rapid Covid Testing
Summary: Summary:The S1 COVID-19 Rapid Antigen test kit developed by Sensing Self Pte. Ltd. can quickly and qualitatively detect the spike glycoprotein of novel coronavirus (SARS-COV-2) in human saliva/sputum/stool samples. It can be used as an aid for COVID-19 diagnosis.Objective:According to the clinical trial plan, the S1 COVID-19 Rapid Antigen test kit or “test reagent”, is to test saliva/sputum/stool samples from healthy subjects and confirmed COVID-19 patients. Test results are compared with another commercial SARS-COV-2 nucleic acid detection kit with CFDA approval, which is defined as the “gold standard”. The sensitivity, specificity, and total agreement rate are used to evaluate the feasibility of the test reagent in clinical applications.Method:A collection of clinical samples were examined by the S1 COVID-19 Rapid Antigen test kit and the gold standard SARS-COV-2 nucleic acid detection kit in parallel, to calculate the clinical sensitivity, clinical specificity and total agreement rate of the test reagent.Results:Compared to the gold standard, the clinical sensitivity of test reagent reached 90.0%, the clinical specificity reached 100.0%, and the total coincidence rate reached 95.0%.Conclusions:The performance of test reagent has a high agreement rate with the gold standard, proving its good feasibility in diagnosing suspected COVID-19 cases.

Proper citation: Shripal Gandhi, Santo Purnama, Keyur Patel, Praveen Sukumara, Dr Rinu R Ravi 2020. SensingSelf S1 Rapid Antigen Test (Saliva/Sputum/Stool). protocols.io https://dx.doi.org/10.17504/protocols.io.bj6ykrfw Copy   

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Authors: Shripal Gandhi, Santo Purnama, Keyur Patel, Praveen Sukumara, Dr Rinu R Ravi
Group: Coronavirus Method Development Community, SensingSelfMission, XPRIZE Rapid Covid Testing
Summary: Summary:The S1 COVID-19 Rapid Antigen test kit developed by Sensing Self Pte. Ltd. can quickly and qualitatively detect the spike glycoprotein of novel coronavirus (SARS-COV-2) in human saliva/sputum/stool samples. It can be used as an aid for COVID-19 diagnosis.Objective:According to the clinical trial plan, the S1 COVID-19 Rapid Antigen test kit or “test reagent”, is to test saliva/sputum/stool samples from healthy subjects and confirmed COVID-19 patients. Test results are compared with another commercial SARS-COV-2 nucleic acid detection kit with CFDA approval, which is defined as the “gold standard”. The sensitivity, specificity, and total agreement rate are used to evaluate the feasibility of the test reagent in clinical applications.Method:A collection of clinical samples were examined by the S1 COVID-19 Rapid Antigen test kit and the gold standard SARS-COV-2 nucleic acid detection kit in parallel, to calculate the clinical sensitivity, clinical specificity and total agreement rate of the test reagent.Results:Compared to the gold standard, the clinical sensitivity of test reagent reached 90.0%, the clinical specificity reached 100.0%, and the total coincidence rate reached 95.0%.Conclusions:The performance of test reagent has a high agreement rate with the gold standard, proving its good feasibility in diagnosing suspected COVID-19 cases.Some of the sample collection steps were adapted from the published protocol:https://www.protocols.io/view/saliva-collection-and-rna-extraction-for-sars-cov-bh6mj9c6

Proper citation: Shripal Gandhi, Santo Purnama, Keyur Patel, Praveen Sukumara, Dr Rinu R Ravi 2020. SensingSelf S1 Rapid Antigen Test (Saliva/Sputum/Stool). protocols.io https://dx.doi.org/10.17504/protocols.io.bpudmns6 Copy   

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Authors: peijun he
Group: Coronavirus Method Development Community, XPRIZE Rapid Covid Testing, Highfield Diagnostics
Summary: Coronavirus Lateral Flow Assay (LFA) operation protocol

Proper citation: peijun he 2020. Coronavirus Lateral Flow Assay (LFA) operation protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.bksjkwcn Copy   

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Authors: peijun he
Group: XPRIZE Rapid Covid Testing, Highfield Diagnostics

Proper citation: peijun he 2020. Highfield Diagnostics COVID19 LFA Protocol v.2. protocols.io https://dx.doi.org/10.17504/protocols.io.bqvqmw5w Copy   

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Authors: yliu
Group: OnsiteGene 1, XPRIZE Rapid Covid Testing
Summary: This OnsiteGene protocol is designed for testing the normal saliva samples with nucleic acid extraction. A normal saliva collection tube is used for the collection. The protocol uses the Star Array® Hi-SenseTM COVID-19 Molecular Testing Kit 1.0 in the one-step real-time RT-qPCR test assay to qualitatively detect RNA from SARS-CoV-2 virus in the human respiratory specimen. It combines the reverse transcription technology and real-time PCR method to provide accurate detection of the SARS-CoV-2 coronavirus. The Star Array® magnetic beads RNA extraction instrument and kit can automatically extract and purify the RNA for 32 samples in 6 minutes. It is ideal for the saliva samples that contain high amount of PCR inhibitors and can't be processed by the direct amplification. The triplex fluorescence design of the kit simultaneously detects the N1 and Orf1ab genes of the virus and the human RNase P gene as an internal control to ensure the sample quality. This protocol uses the Star Array® XDiveTM Superfast Real-Time PCR instrument to perform 40-cycle PCR in 8 minutes, and can test up to 16 samples or controls in each run. The total protocol time from sample collection to data interpretation is less than 15 minutes.

Proper citation: yliu 2020. OnsiteGene 4 Protocol Saliva Extract. protocols.io https://dx.doi.org/10.17504/protocols.io.bkujkwun Copy   

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Authors: Mahe Raccaud, Manon Garzuel, Joerg Ziegler, Luc Gervais
Group: Coronavirus Method Development Community, XPRIZE Rapid Covid Testing
Summary: Our capillary-driven microfluidic chips automatically detect multiple SARS-CoV-2 antigens after the addition of a few drops of saliva, triggering a cascade of events powered by capillary forces. We integrate biochemical reagents directly on the microfluidic chips, avoiding the need for peripheral equipment for fluid control. Thus, detection of SARS-CoV-2 can be done at the patient’s side by simply adding sample to the chips, with minimal hands-on time, faster results, and increased sensitivity of fluorescence assays. The use of saliva as a sample matrix avoids painful sample-collection procedures and the need for trained medical staff.

Proper citation: Mahe Raccaud, Manon Garzuel, Joerg Ziegler, Luc Gervais 2020. 1DROP SARS-CoV-2 antigen test. protocols.io https://dx.doi.org/10.17504/protocols.io.bk38kyrw Copy   

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Authors: John Daniels
Group: Coronavirus Method Development Community, Front Line Technologies, XPRIZE Rapid Covid Testing, Team Exhaled Breath Condensate

Proper citation: John Daniels 2020. Mask-Based Covid-19 testing system using Exhaled Breath Condensate. protocols.io https:// Copy   

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Authors: Guanhua Xun, Huimin Zhao, stlane2
Group: XPRIZE Rapid Covid Testing, SPOT

Proper citation: Guanhua Xun, Huimin Zhao, stlane2 2020. SPOT1 assay. protocols.io https://dx.doi.org/10.17504/protocols.io.bk25kyg6 Copy   

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Authors: Guy Aidelberg, Rachel Aronoff
Group: Coronavirus Method Development Community, Reclone.org (The Reagent Collaboration Network), XPRIZE Rapid Covid Testing, Guy Aidelberg
Summary: A protocol for freeze-drying (Lyophilization) of CoronaDetective tests, and more generally any QUASR RT-LAMP reaction.The amounts here are for one 96 well standard PCR plate and for reactions This can be scaled for any amount and size.

Proper citation: Guy Aidelberg, Rachel Aronoff 2020. Freeze-drying (Lyophilization) and manufacturing of Corona Detective assay. protocols.io https://dx.doi.org/10.17504/protocols.io.bpv4mn8w Copy   

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Authors: Rushna Quddus, Kaitlynn Snyder, Lauren Altman, Laura Philips, David G. Grier, Andrew D. Hollingsworth, Kent Kirshenbaum
Group: Coronavirus Method Development Community, Grier Group, HolographicDiagnostics, XPRIZE Rapid Covid Testing
Summary: This protocol describes the steps required to perform a holographic immunoassay for SARS-CoV-2 whole virus particles. The assay uses an xSight holographic particle characterization instrument (Spheryx, Inc.) to monitor the diameter of specifically functionalized probe beads. The diameter of the beads increases by a few nanometers as targets bind to the surface. xSight detects and reports this change, yielding an estimate for the concentration of analytes in the sample. The method for holographic immunoassays (applied to antibody binding assays) is described inY. Zagzag, M. F. Soddu, A. D. Hollingsworth and D. G. Grier, Holographic molecular binding assays, Scientific Reports 10, 1932 (2020) and K. Snyder, R. Quddus, A. D. Hollingsworth and K. Kirshenbaum, Holographic Immunoassays: Direct Detection of Antibodies Binding to Colloidal Spheres, submitted for publication (2020).

Proper citation: Rushna Quddus, Kaitlynn Snyder, Lauren Altman, Laura Philips, David G. Grier, Andrew D. Hollingsworth, Kent Kirshenbaum 2020. HolographicDiagnostics: Automated Virus Binding Assay. protocols.io https://dx.doi.org/10.17504/protocols.io.bkpgkvjw Copy   

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Authors: Ladislav Nevery
Group: Coronavirus Method Development Community, XPRIZE Rapid Covid Testing, Ladislav Nevery
Summary: By passing saliva sample thru 200nm filter to remove particles larger than 120nm covid 19 virus particles and concentrating resulting fluid to very small 1 mm2 area of 100nm filter. Pores in this small area of100nm filter should be clogged up fast raising filter air resistance. Due to high virus density 5.2 log10 ml saliva of infected person should clogg filter much faster than healthy person. Test is setup in such way that act of compressor automatically turning off after reaching threshold PSI value will indicate high count of particles larger than 100nm of being present in measured filter area.

Proper citation: Ladislav Nevery 2020. covid 19 indirect detection thru rise of 100nm filter air resistance . protocols.io https://dx.doi.org/10.17504/protocols.io.bkznkx5e Copy   

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Authors: John Warner, Rebecca Silveston, Delphine Dean
Group: Coronavirus Method Development Community, XPRIZE Rapid Covid Testing, Accessible Health

Proper citation: John Warner, Rebecca Silveston, Delphine Dean 2020. Accessible Health Sentinal Test Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.bk5wky7e Copy   

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Authors: Tho Ho
Group: VMMU, XPRIZE Rapid Covid Testing

Proper citation: Tho Ho 2020. Pooling protocol using AMPHABIO HT-HiThroughput PCR COVID-19 Kit. protocols.io https://dx.doi.org/10.17504/protocols.io.bke4ktgw Copy   

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Authors: Brian Glenn St Hilaire
Group: XPRIZE Rapid Covid Testing

Proper citation: Brian Glenn St Hilaire 2020. POLAR Express: Pathogen-Oriented Low-cost Assembly & Re-sequencing. protocols.io https://dx.doi.org/10.17504/protocols.io.bhv5j686 Copy   

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Authors: huan.jia , patthara
Group: XPRIZE Rapid Covid Testing, Poseidon

Proper citation: huan.jia , patthara 2020. User Protocol for SaliCoV Detection Kit. protocols.io https://dx.doi.org/10.17504/protocols.io.bkymkxu6 Copy   

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Authors: Tho Ho
Group: Coronavirus Method Development Community, VMMU, XPRIZE Rapid Covid Testing
Summary: This protocol shows procedure for sample handling and detection of SARS-CoV-2 using the AMPHABIO HT-HiThroughput PCR COVID-19 Kit.

Proper citation: Tho Ho 2020. AMPHABIO HT-HiThroughput PCR COVID-19 Kit Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.bqdvms66 Copy   

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Authors: Qianxin Wu, Chenqu Suo, Tom Brown, Tengyao Wang, Sarah Teichmann, Andrew Bassett
Group: Coronavirus Method Development Community, XPRIZE Rapid Covid Testing

Proper citation: Qianxin Wu, Chenqu Suo, Tom Brown, Tengyao Wang, Sarah Teichmann, Andrew Bassett 2020. INSIGHT: a population scale COVID-19 testing strategy combining point-of-care diagnosis with centralised high-throughput sequencing. protocols.io https:// Copy   

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Authors: Anne Berking
Group: XPRIZE Rapid Covid Testing
Summary: The "Rapid Ag test based on Alpaca Nanobodies" is a test developed with single domain antibodies. This type of antibody is capable of effectively recognize the Spike protein on the surface of SARS-CoV2 leading a signal that is 10 times stronger than conventional antibodies. It takes only 8 minutes from sample to result. The procedures for using this test were designed to ensure an easy use and interpretation, making it a quick and simple test.

Proper citation: Anne Berking 2020. Alpaca Nanobodies Rapid Ag test. protocols.io https://dx.doi.org/10.17504/protocols.io.bk3xkypn Copy   

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Authors: Mario Thomas, Jasmine Sollen, Natalia Ivanova, Heidi Abdilla, Reda Fayek, Michelle Feng, Stephanie Lim, Amanda Naaum
Group: XPRIZE Rapid Covid Testing, SANATA
Summary: This procedure outlines the protocol using any swab suitable for a SARS-CoV-2 sample collection. The purpose of this test is to detect low levels of SARS-CoV-2 antigen at a higher sensitivity. Precision Biomonitoring Inc. developed an ultra-rapid digital, disposable, highly-sensitive and inexpensive testing device used for screening purposes. The mobile app complementary to this medical device is connected through Bluetooth. Using this innovation, the patients can be tested at the point-of-care (POC), and obtain qualitative results. It is recommended that positive results should be confirmed by RT-PCR.

Proper citation: Mario Thomas, Jasmine Sollen, Natalia Ivanova, Heidi Abdilla, Reda Fayek, Michelle Feng, Stephanie Lim, Amanda Naaum 2020. XPRIZE SANATA Protocol for Swab LFIA Test. protocols.io https://dx.doi.org/10.17504/protocols.io.bkzjkx4n Copy   

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Authors: Randy True
Group: FloodLAMP, XPRIZE Rapid Covid Testing
Summary: FloodLAMP uses an assay chemistry developed by Brian Rabe and Prof. Connie Cepko at The Cepko Lab at Harvard Medical School. "Rabe Cepko," as it's known, uses an ultra cheap, streamlined front end (sample inactivation and RNA purification+concentration). This protocol has been chosen by many groups, from pharmaceutical companies to researchers working in the developing world. The same aspects that make it ideal for Africa also make it ideal for America—it's cheap, works well, and can be implemented by any basic chemistry or biology lab.This assay was validated clinically in May by researchers at Mass General Hospital. In addition, the key amplification reagent (NEB Colorimetric LAMP Master Mix) is the same product used in the FDA-approved test by Color Genomics, who does about 5K tests per day in the SF Bay Area. Our current preferred version incorporates a transfer of the nucleic acid bound silica in ethanol to the PCR tubes (strips or plates) where the LAMP reaction is carried out. The pellet resuspends easily in the 80% ethanol and—once in the PCR tubes—can be quickly dried on a heat block. Then they are ready for addition of the 1X LAMP Reaction Mix. We have used alternatives to this method, including an elution of the nucleic acid and also a resuspension of the pellet in water or 1X PBS, then addition to the LAMP reaction. With the LAMP Master Mix (NEB 1804), the assay uses 2 prepared solutions: a NaI Binding Solution and the Glass Milk (also called "prepared silica"). On our website are our protocols in worksheet form as we use in the lab. This and more information will be coming soon.

Proper citation: Randy True 2020. FloodLAMP LAMP Assay Protocol v4.1 (0.5ml, Strips, Multichannel). protocols.io https://dx.doi.org/10.17504/protocols.io.bkvnkw5e Copy   

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