Rapid isothermal point-of-care test for screening of SARS-CoV-2 (COVID-19)
Rapid on-site diagnosis of emerging pathogens is key for early identification of infected individuals and for prevention of further spreading in a population. Currently available molecular diagnostic tests are instrument-based whereas rapid antibody and antigen tests are often not sufficiently sensi...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-01-01
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| Series: | Aspects of Molecular Medicine |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2949688823000023 |
| _version_ | 1797237869532676096 |
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| author | Jean-Marc Zingg Yu-Ping Yang Spencer Seely Pratibha Joshi Md Harun Or Roshid Fabiola Iribarren Latasa Gregory O'Connor Jennifer Alfaro Eduardo Riquelme Sebastian Bernales Emre Dikici Sapna Deo Sylvia Daunert |
| author_facet | Jean-Marc Zingg Yu-Ping Yang Spencer Seely Pratibha Joshi Md Harun Or Roshid Fabiola Iribarren Latasa Gregory O'Connor Jennifer Alfaro Eduardo Riquelme Sebastian Bernales Emre Dikici Sapna Deo Sylvia Daunert |
| author_sort | Jean-Marc Zingg |
| collection | DOAJ |
| description | Rapid on-site diagnosis of emerging pathogens is key for early identification of infected individuals and for prevention of further spreading in a population. Currently available molecular diagnostic tests are instrument-based whereas rapid antibody and antigen tests are often not sufficiently sensitive for detection in pre-symptomatic subjects. There is a need for rapid point of care molecular screening tests that can be easily adapted to emerging pathogens and are selective, sensitive, reliable in different settings around the world. We have developed a simple, rapid (<30 min), and inexpensive test for SARS-CoV-2 that is based on combination of isothermal reverse transcription recombinase polymerase amplification (RT-RPA) using modified primers and visual detection with paper-based microfluidics. Our test (CoRapID) is specific for SARS-CoV-2 (alpha to omicron variants) and does not detect other coronaviruses and pathogens by in silico and in vitro analysis. A two-step test protocol was developed with stable lyophilized reagents that reduces handling by using portable and disposable components (droppers, microapplicators/swabs, paper-strips). After optimization of assay components and conditions, we have achieved a limit of detection (LoD) of 1 copy/reaction by adding a blocking primer to the lateral flow assay. Using a set of 138 clinical samples, a sensitivity of 88.1% (P < 0.05, CI: 78.2–93.8%) and specificity of 93.9% (P < 0.05, CI: 85.4–97.6%) was determined. The lack of need for instrumentation for our CoRapID makes it an ideal on-site primary screening tool for local hospitals, doctors’ offices, senior homes, workplaces, and in remote settings around the world that often do not have access to clinical laboratories. |
| first_indexed | 2024-04-24T17:26:36Z |
| format | Article |
| id | doaj.art-161bd1c89ecf4cc69b200052ec6fe175 |
| institution | Directory Open Access Journal |
| issn | 2949-6888 |
| language | English |
| last_indexed | 2024-04-24T17:26:36Z |
| publishDate | 2023-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Aspects of Molecular Medicine |
| spelling | doaj.art-161bd1c89ecf4cc69b200052ec6fe1752024-03-28T06:39:48ZengElsevierAspects of Molecular Medicine2949-68882023-01-011100002Rapid isothermal point-of-care test for screening of SARS-CoV-2 (COVID-19)Jean-Marc Zingg0Yu-Ping Yang1Spencer Seely2Pratibha Joshi3Md Harun Or Roshid4Fabiola Iribarren Latasa5Gregory O'Connor6Jennifer Alfaro7Eduardo Riquelme8Sebastian Bernales9Emre Dikici10Sapna Deo11Sylvia Daunert12Department of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami, Miami, FL, 33136-6129, USA; Corresponding author. Department of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami, Miami, FL, 33136-6129, USA.Department of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami, Miami, FL, 33136-6129, USADepartment of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami, Miami, FL, 33136-6129, USADepartment of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami, Miami, FL, 33136-6129, USADepartment of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami, Miami, FL, 33136-6129, USA; Department of Chemistry, University of Miami, Miami, FL, 33146, USADepartment of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami, Miami, FL, 33136-6129, USA; Universidad Francisco de Vitoria, Madrid, SpainDepartment of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami, Miami, FL, 33136-6129, USAMerken Biotech SpA, Zañartu, 1482, Santiago, ChileMerken Biotech SpA, Zañartu, 1482, Santiago, ChileMerken Biotech SpA, Zañartu, 1482, Santiago, Chile; Centro Ciencia & Vida, Zañartu, 1482, Santiago, ChileDepartment of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami, Miami, FL, 33136-6129, USA; Dr. JT Macdonald Foundation Biomedical Nanotechnology Institute, University of Miami, Miami, FL, 33136-6129, USADepartment of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami, Miami, FL, 33136-6129, USA; Dr. JT Macdonald Foundation Biomedical Nanotechnology Institute, University of Miami, Miami, FL, 33136-6129, USA; Corresponding author. Department of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami, Miami, FL, 33136-6129, USA.Department of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami, Miami, FL, 33136-6129, USA; Dr. JT Macdonald Foundation Biomedical Nanotechnology Institute, University of Miami, Miami, FL, 33136-6129, USA; University of Miami Clinical and Translational Science Institute, University of Miami, Miami, FL, 33136-6129, USA; Corresponding author. Department of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami, Miami, FL, 33136-6129, USARapid on-site diagnosis of emerging pathogens is key for early identification of infected individuals and for prevention of further spreading in a population. Currently available molecular diagnostic tests are instrument-based whereas rapid antibody and antigen tests are often not sufficiently sensitive for detection in pre-symptomatic subjects. There is a need for rapid point of care molecular screening tests that can be easily adapted to emerging pathogens and are selective, sensitive, reliable in different settings around the world. We have developed a simple, rapid (<30 min), and inexpensive test for SARS-CoV-2 that is based on combination of isothermal reverse transcription recombinase polymerase amplification (RT-RPA) using modified primers and visual detection with paper-based microfluidics. Our test (CoRapID) is specific for SARS-CoV-2 (alpha to omicron variants) and does not detect other coronaviruses and pathogens by in silico and in vitro analysis. A two-step test protocol was developed with stable lyophilized reagents that reduces handling by using portable and disposable components (droppers, microapplicators/swabs, paper-strips). After optimization of assay components and conditions, we have achieved a limit of detection (LoD) of 1 copy/reaction by adding a blocking primer to the lateral flow assay. Using a set of 138 clinical samples, a sensitivity of 88.1% (P < 0.05, CI: 78.2–93.8%) and specificity of 93.9% (P < 0.05, CI: 85.4–97.6%) was determined. The lack of need for instrumentation for our CoRapID makes it an ideal on-site primary screening tool for local hospitals, doctors’ offices, senior homes, workplaces, and in remote settings around the world that often do not have access to clinical laboratories.http://www.sciencedirect.com/science/article/pii/S2949688823000023CoronavirusPoint of care (PoC) testLateral flow assay (LFA)Recombinase polymerase amplification (RPA)Isothermal amplificationReverse transcription recombinase polymerase amplification (RT-RPA) |
| spellingShingle | Jean-Marc Zingg Yu-Ping Yang Spencer Seely Pratibha Joshi Md Harun Or Roshid Fabiola Iribarren Latasa Gregory O'Connor Jennifer Alfaro Eduardo Riquelme Sebastian Bernales Emre Dikici Sapna Deo Sylvia Daunert Rapid isothermal point-of-care test for screening of SARS-CoV-2 (COVID-19) Aspects of Molecular Medicine Coronavirus Point of care (PoC) test Lateral flow assay (LFA) Recombinase polymerase amplification (RPA) Isothermal amplification Reverse transcription recombinase polymerase amplification (RT-RPA) |
| title | Rapid isothermal point-of-care test for screening of SARS-CoV-2 (COVID-19) |
| title_full | Rapid isothermal point-of-care test for screening of SARS-CoV-2 (COVID-19) |
| title_fullStr | Rapid isothermal point-of-care test for screening of SARS-CoV-2 (COVID-19) |
| title_full_unstemmed | Rapid isothermal point-of-care test for screening of SARS-CoV-2 (COVID-19) |
| title_short | Rapid isothermal point-of-care test for screening of SARS-CoV-2 (COVID-19) |
| title_sort | rapid isothermal point of care test for screening of sars cov 2 covid 19 |
| topic | Coronavirus Point of care (PoC) test Lateral flow assay (LFA) Recombinase polymerase amplification (RPA) Isothermal amplification Reverse transcription recombinase polymerase amplification (RT-RPA) |
| url | http://www.sciencedirect.com/science/article/pii/S2949688823000023 |
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