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dc.contributor.authorTakahashi, Masateru
dc.contributor.authorTehseen, Muhammad
dc.contributor.authorSalunke, Rahul Pandurang
dc.contributor.authorTakahashi, Etsuko
dc.contributor.authorMfarrej, Sara
dc.contributor.authorSobhy, Mohamed Abdelmaboud
dc.contributor.authorAlhamlan, Fatimah S.
dc.contributor.authorHala, Sharif
dc.contributor.authorRamos Mandujano, Gerardo
dc.contributor.authorAl-Qahtani, Ahmed A.
dc.contributor.authorAlofi, Fadwa S.
dc.contributor.authorAlsomali, Afrah
dc.contributor.authorHashem, Anwar M.
dc.contributor.authorKhogeer, Asim
dc.contributor.authorAlmontashiri, Naif A. M.
dc.contributor.authorLee, Jae Man
dc.contributor.authorMon, Hiroaki
dc.contributor.authorSakashita, Kosuke
dc.contributor.authorLi, Mo
dc.contributor.authorKusakabe, Takahiro
dc.contributor.authorPain, Arnab
dc.contributor.authorHamdan, Samir
dc.date.accessioned2021-03-24T13:09:35Z
dc.date.available2021-03-24T13:09:35Z
dc.date.issued2021-03-15
dc.date.submitted2020-11-19
dc.identifier.citationTakahashi, M., Tehseen, M., Salunke, R., Takahashi, E., Mfarrej, S., Sobhy, M. A., … Hamdan, S. M. (2021). Quick and Easy Assembly of a One-Step qRT-PCR Kit for COVID-19 Diagnostics Using In-House Enzymes. ACS Omega. doi:10.1021/acsomega.0c05635
dc.identifier.issn2470-1343
dc.identifier.issn2470-1343
dc.identifier.doi10.1021/acsomega.0c05635
dc.identifier.urihttp://hdl.handle.net/10754/668245
dc.description.abstractOne-step reverse-transcription quantitative polymerase chain reaction (qRT-PCR) is the most widely applied method for COVID-19 diagnostics. Notwithstanding the facts that one-step qRT-PCR is well suited for the diagnosis of COVID-19 and that there are many commercially available one-step qRT-PCR kits in the market, their high cost and unavailability due to airport closures and shipment restriction became a major bottleneck that had driven the desire to produce the key components of such kits locally. Here, we provide a simple, economical, and powerful one-step qRT-PCR kit based on patent-free, specifically tailored versions of Moloney murine leukemia virus reverse transcriptase and Thermus aquaticus DNA polymerase and termed R3T (Rapid Research Response Team) one-step qRT-PCR. We also demonstrate the robustness of our enzyme production strategies and provide the optimal reaction conditions for their efficient augmentation in a one-step approach. Our kit was routinely able to reliably detect as low as 10 copies of the synthetic RNAs of SARS-CoV-2. More importantly, our kit successfully detected COVID-19 in clinical samples of broad viral titers with similar reliability and selectivity to that of the Invitrogen SuperScript III Platinum One-step qRT-PCR and TaqPath one-step RT-qPCR kits. Overall, our kit has shown robust performance in both laboratory settings and the Saudi Ministry of Health-approved testing facility.
dc.description.sponsorshipThis research was funded by baseline funding from KAUST to S.M.H. and COVID-19 response initiative by the Vice President of research at KAUST. The clinical COVID-19 samples were collected as part of KAUST baseline funding (BAS/1/1020-01-01) to AP and the R3T initiative by the the Vice President of research at KAUST. The authors declare no conflicts of interest.
dc.publisherAmerican Chemical Society (ACS)
dc.relation.urlhttps://pubs.acs.org/doi/10.1021/acsomega.0c05635
dc.rightsThis document is the Accepted Manuscript version of a Published Work that appeared in final form in ACS Omega, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://pubs.acs.org/doi/10.1021/acsomega.0c05635.
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.titleQuick and Easy Assembly of a One-Step qRT-PCR Kit for COVID-19 Diagnostics Using In-House Enzymes
dc.typeArticle
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Division
dc.contributor.departmentBioscience Program
dc.contributor.departmentLaboratory of DNA Replication and Recombination
dc.contributor.departmentPathogen Genomics Laboratory
dc.contributor.departmentProteomics, protein expression & cytomet
dc.identifier.journalACS Omega
dc.eprint.versionPublisher's Version/PDF
dc.contributor.institutionDepartment of Infection and Immunity, King Faisal Specialist Hospital and Research Center, Riyadh 11211, Saudi Arabia
dc.contributor.institutionKing Saud Bin Abdulaziz University of Health Sciences, Jeddah 22384, Saudi Arabia
dc.contributor.institutionKing Abdullah International Medical Research Centre, Jeddah, Makkah, Ministry of National Guard Health Affairs, Jeddah, Makkah 22384, Saudi Arabia
dc.contributor.institutionInfectious Diseases Department, King Fahad Hospital, Madinah 3177, Saudi Arabia
dc.contributor.institutionKing Abdullah Medical Complex (KAMC), Jeddah 23816, Saudi Arabia
dc.contributor.institutionVaccines and Immunotherapy Unit, King Fahd Medical Research Center; King Abdulaziz University, Jeddah, Saudi Arabia
dc.contributor.institutionDepartment of Medical Microbiology and Parasitology, Faculty of Medicine, King Abdulaziz University, Jeddah 21589, Saudi Arabia
dc.contributor.institutionPlan and Research Department, General Directorate of Health Affairs Makkah Region, MOH Mecca 24321, Saudi Arabia
dc.contributor.institutionCollege of Applied Medical Sciences, Taibah University, Madinah 41311, Saudi Arabia
dc.contributor.institutionCenter for Genetics and Inherited Diseases, Taibah University, Madinah 42353, Saudi Arabia
dc.contributor.institutionLaboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Motooka 744, Nishi-ku, Fukuoka 819-0395, Japan
kaust.personTakahashi, Masateru
kaust.personTehseen, Muhammad
kaust.personSalunke, Rahul Pandurang
kaust.personTakahashi, Etsuko
kaust.personMfarrej, Sara
kaust.personSobhy, Mohamed Abdelmaboud
kaust.personHala, Sharif
kaust.personRamos Mandujano, Gerardo
kaust.personSakashita, Kosuke
kaust.personLi, Mo
kaust.personPain, Arnab
kaust.personHamdan, Samir
kaust.grant.numberBAS/1/1020-01-01
dc.date.accepted2021-03-01
refterms.dateFOA2021-03-24T13:10:52Z
kaust.acknowledged.supportUnitKAUST baseline funding


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This document is the Accepted Manuscript version of a Published Work that appeared in final form in ACS Omega, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://pubs.acs.org/doi/10.1021/acsomega.0c05635.
Except where otherwise noted, this item's license is described as This document is the Accepted Manuscript version of a Published Work that appeared in final form in ACS Omega, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://pubs.acs.org/doi/10.1021/acsomega.0c05635.