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R3T (Rapid Research Response Team) One-step RT-qPCR kit for COVID-19 diagnostic using in-house enzymes

dc.contributor.authorTakahashi, Masateru
dc.contributor.authorTehseen, Muhammad
dc.contributor.authorSalunke, Rahul
dc.contributor.authorTakahashi, Etsuko
dc.contributor.authorMfarrej, Sara
dc.contributor.authorSobhy, Mohamed Abdelmaboud
dc.contributor.authorAlhamlan, Fatimah
dc.contributor.authorHala, Sharif
dc.contributor.authorMandujano, Gerardo R.
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.accessioned2020-08-18T13:08:01Z
dc.date.available2020-08-18T13:08:01Z
dc.date.issued2020-08-04
dc.identifier.citationTakahashi, M., Tehseen, M., Salunke, R., Takahashi, E., Mfarrej, S., Sobhy, M. A., … Hamdan, S. M. (2020). R3T (Rapid Research Response Team) One-step RT-qPCR kit for COVID-19 diagnostic using in-house enzymes. doi:10.1101/2020.07.31.20165704
dc.identifier.doi10.1101/2020.07.31.20165704
dc.identifier.urihttp://hdl.handle.net/10754/664657
dc.description.abstractOne-step RT-qPCR is the most widely applied method for COVID-19 diagnostics. Designing in-house one-step RT-qPCR kits is restricted by the patent-rights for the production of enzymes and the lack of information about the components of the commercial kits. Here, we provide a simple, economical, and powerful one-step RT-qPCR kit based on patent-free, specifically-tailored versions of Moloney Murine Leukemia Virus Reverse Transcriptase and Thermus aquaticus DNA polymerase termed the R3T (Rapid Research Response Team) One-step RT-qPCR. Our kit was routinely able to reliably detect as low as 10 copies of the synthetic RNAs of the SARS-CoV-2. More importantly, our kit successfully detected COVID-19 in clinical samples of broad viral titers with similar reliability and selectivity as that of the Invitrogen SuperScript III Platinum One-step RT-qPCR and TaqPath 1-Step RT-qPCR kits. Overall, our kit has shown robust performance in both of 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 COVID19 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.publisherCold Spring Harbor Laboratory
dc.relation.urlhttp://medrxiv.org/lookup/doi/10.1101/2020.07.31.20165704
dc.rightsArchived with thanks to Cold Spring Harbor Laboratory
dc.titleR3T (Rapid Research Response Team) One-step RT-qPCR kit for COVID-19 diagnostic using in-house enzymes
dc.typePreprint
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.departmentPathogen Genomics Laboratory, BESE Division, King Abdullah University of Science and Technology (KAUST), Thuwal 23955-6900, Saudi Arabia.
dc.contributor.departmentProteomics and Protein Expression
dc.contributor.departmentStem Cell and Regenration Laboratory. Biological and Environmental Sciences and Engineering Division, King Abdullah University of Science and Technology (KAUST), Thuwal 23955-6900, Saudi Arabia.
dc.eprint.versionPre-print
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, Saudi Arabia.
dc.contributor.institutionKing Abdullah International Medical Research Centre, Jeddah, Makkah, Ministry of National Guard Health Affairs, Jeddah, Makkah, Saudi Arabia.
dc.contributor.institutionInfectious Diseases Department, King Fahad Hospital, Madinah, Saudi Arabia.
dc.contributor.institutionKing Abdullah Medical Complex (KAMC), Jeddah, Saudi Arabia.
dc.contributor.institutionVaccines and Immunotherapy Unit, King Fahd Medical Research Center; King Abdulaziz University, Jeddah, Saudi Arabia; Department of Medical Microbiology and Parasitology, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia.
dc.contributor.institutionPlan and Research Department, General Directorate of Health Affairs Makkah Region, MOH, Saudi Arabia.
dc.contributor.institutionCollege of Applied Medical Sciences, Taibah University, Madinah, Saudi Arabia; Center for Genetics and Inherited Diseases, Taibah University, Madinah, 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
kaust.personTakahashi, Etsuko
kaust.personMfarrej, Sara
kaust.personSobhy, Mohamed Abdelmaboud
kaust.personHala, Sharif
kaust.personMandujano, Gerardo R.
kaust.personSakashita, Kosuke
kaust.personLi, Mo
kaust.personPain, Arnab
kaust.personHamdan, Samir
kaust.grant.numberBAS/1/1020-01-01
refterms.dateFOA2020-08-18T13:10:57Z
kaust.acknowledged.supportUnitKAUST baseline fund


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