R3T (Rapid Research Response Team) One-step RT-qPCR kit for COVID-19 diagnostic using in-house enzymes
Sobhy, Mohamed Abdelmaboud
Mandujano, Gerardo R.
Al-Qahtani, Ahmed A.
Alofi, Fadwa S.
Hashem, Anwar M.
Almontashiri, Naif A. M.
Lee, Jae Man
KAUST DepartmentBiological and Environmental Sciences and Engineering (BESE) Division
Laboratory of DNA Replication and Recombination
Pathogen Genomics Laboratory
Pathogen Genomics Laboratory, BESE Division, King Abdullah University of Science and Technology (KAUST), Thuwal 23955-6900, Saudi Arabia.
Proteomics and Protein Expression
Stem Cell and Regenration Laboratory. Biological and Environmental Sciences and Engineering Division, King Abdullah University of Science and Technology (KAUST), Thuwal 23955-6900, Saudi Arabia.
KAUST Grant NumberBAS/1/1020-01-01
Permanent link to this recordhttp://hdl.handle.net/10754/664657
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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.
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
SponsorsThis 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.
PublisherCold Spring Harbor Laboratory