Evaluation of mobile real-time polymerase chain reaction tests for the detection of severe acute respiratory syndrome coronavirus 2

Abstract The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), calls for prompt and accurate diagnosis and rapid turnaround time for test results to limit transmission. Here, we evaluated two independent molecular assays, the Biomem...

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Autores principales: Chukwunonso Onyilagha, Henna Mistry, Peter Marszal, Mathieu Pinette, Darwyn Kobasa, Nikesh Tailor, Yohannes Berhane, Charles Nfon, Bradley Pickering, Samira Mubareka, David Bulir, Sylvia Chong, Robert Kozak, Aruna Ambagala
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Publicado: Nature Portfolio 2021
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spelling oai:doaj.org-article:a17bab77e5ff48acb074e774c53b39f72021-12-02T13:41:34ZEvaluation of mobile real-time polymerase chain reaction tests for the detection of severe acute respiratory syndrome coronavirus 210.1038/s41598-021-88625-62045-2322https://doaj.org/article/a17bab77e5ff48acb074e774c53b39f72021-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-88625-6https://doaj.org/toc/2045-2322Abstract The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), calls for prompt and accurate diagnosis and rapid turnaround time for test results to limit transmission. Here, we evaluated two independent molecular assays, the Biomeme SARS-CoV-2 test, and the Precision Biomonitoring TripleLock SARS-CoV-2 test on a field-deployable point-of-care real-time PCR instrument, Franklin three9, in combination with Biomeme M1 Sample Prep Cartridge Kit for RNA 2.0 (M1) manual extraction system for rapid, specific, and sensitive detection of SARS-COV-2 in cell culture, human, and animal clinical samples. The Biomeme SARS-CoV-2 assay, which simultaneously detects two viral targets, the orf1ab and S genes, and the Precision Biomonitoring TripleLock SARS-CoV-2 assay that targets the 5′ untranslated region (5′ UTR) and the envelope (E) gene of SARS-CoV-2 were highly sensitive and detected as low as 15 SARS-CoV-2 genome copies per reaction. In addition, the two assays were specific and showed no cross-reactivity with Middle Eastern respiratory syndrome coronavirus (MERS-CoV), infectious bronchitis virus (IBV), porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis (TGE) virus, and other common human respiratory viruses and bacterial pathogens. Also, both assays were highly reproducible across different operators and instruments. When used to test animal samples, both assays equally detected SARS-CoV-2 genetic materials in the swabs from SARS-CoV-2-infected hamsters. The M1 lysis buffer completely inactivated SARS-CoV-2 within 10 min at room temperature enabling safe handling of clinical samples. Collectively, these results show that the Biomeme and Precision Biomonitoring TripleLock SARS-CoV-2 mobile testing platforms could reliably and promptly detect SARS-CoV-2 in both human and animal clinical samples in approximately an hour and can be used in remote areas or health care settings not traditionally serviced by a microbiology laboratory.Chukwunonso OnyilaghaHenna MistryPeter MarszalMathieu PinetteDarwyn KobasaNikesh TailorYohannes BerhaneCharles NfonBradley PickeringSamira MubarekaDavid BulirSylvia ChongRobert KozakAruna AmbagalaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution DOAJ
collection DOAJ
language EN
topic Medicine
R
Science
Q
spellingShingle Medicine
R
Science
Q
Chukwunonso Onyilagha
Henna Mistry
Peter Marszal
Mathieu Pinette
Darwyn Kobasa
Nikesh Tailor
Yohannes Berhane
Charles Nfon
Bradley Pickering
Samira Mubareka
David Bulir
Sylvia Chong
Robert Kozak
Aruna Ambagala
Evaluation of mobile real-time polymerase chain reaction tests for the detection of severe acute respiratory syndrome coronavirus 2
description Abstract The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), calls for prompt and accurate diagnosis and rapid turnaround time for test results to limit transmission. Here, we evaluated two independent molecular assays, the Biomeme SARS-CoV-2 test, and the Precision Biomonitoring TripleLock SARS-CoV-2 test on a field-deployable point-of-care real-time PCR instrument, Franklin three9, in combination with Biomeme M1 Sample Prep Cartridge Kit for RNA 2.0 (M1) manual extraction system for rapid, specific, and sensitive detection of SARS-COV-2 in cell culture, human, and animal clinical samples. The Biomeme SARS-CoV-2 assay, which simultaneously detects two viral targets, the orf1ab and S genes, and the Precision Biomonitoring TripleLock SARS-CoV-2 assay that targets the 5′ untranslated region (5′ UTR) and the envelope (E) gene of SARS-CoV-2 were highly sensitive and detected as low as 15 SARS-CoV-2 genome copies per reaction. In addition, the two assays were specific and showed no cross-reactivity with Middle Eastern respiratory syndrome coronavirus (MERS-CoV), infectious bronchitis virus (IBV), porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis (TGE) virus, and other common human respiratory viruses and bacterial pathogens. Also, both assays were highly reproducible across different operators and instruments. When used to test animal samples, both assays equally detected SARS-CoV-2 genetic materials in the swabs from SARS-CoV-2-infected hamsters. The M1 lysis buffer completely inactivated SARS-CoV-2 within 10 min at room temperature enabling safe handling of clinical samples. Collectively, these results show that the Biomeme and Precision Biomonitoring TripleLock SARS-CoV-2 mobile testing platforms could reliably and promptly detect SARS-CoV-2 in both human and animal clinical samples in approximately an hour and can be used in remote areas or health care settings not traditionally serviced by a microbiology laboratory.
format article
author Chukwunonso Onyilagha
Henna Mistry
Peter Marszal
Mathieu Pinette
Darwyn Kobasa
Nikesh Tailor
Yohannes Berhane
Charles Nfon
Bradley Pickering
Samira Mubareka
David Bulir
Sylvia Chong
Robert Kozak
Aruna Ambagala
author_facet Chukwunonso Onyilagha
Henna Mistry
Peter Marszal
Mathieu Pinette
Darwyn Kobasa
Nikesh Tailor
Yohannes Berhane
Charles Nfon
Bradley Pickering
Samira Mubareka
David Bulir
Sylvia Chong
Robert Kozak
Aruna Ambagala
author_sort Chukwunonso Onyilagha
title Evaluation of mobile real-time polymerase chain reaction tests for the detection of severe acute respiratory syndrome coronavirus 2
title_short Evaluation of mobile real-time polymerase chain reaction tests for the detection of severe acute respiratory syndrome coronavirus 2
title_full Evaluation of mobile real-time polymerase chain reaction tests for the detection of severe acute respiratory syndrome coronavirus 2
title_fullStr Evaluation of mobile real-time polymerase chain reaction tests for the detection of severe acute respiratory syndrome coronavirus 2
title_full_unstemmed Evaluation of mobile real-time polymerase chain reaction tests for the detection of severe acute respiratory syndrome coronavirus 2
title_sort evaluation of mobile real-time polymerase chain reaction tests for the detection of severe acute respiratory syndrome coronavirus 2
publisher Nature Portfolio
publishDate 2021
url https://doaj.org/article/a17bab77e5ff48acb074e774c53b39f7
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