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

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Authors
Takahashi, Masateru cc
Tehseen, Muhammad cc
Salunke, Rahul
Takahashi, Etsuko cc
Mfarrej, Sara
Sobhy, Mohamed Abdelmaboud
Alhamlan, Fatimah
Hala, Sharif cc
Mandujano, Gerardo R.
Al-Qahtani, Ahmed A.
Alofi, Fadwa S.
Alsomali, Afrah
Hashem, Anwar M.
Khogeer, Asim
Almontashiri, Naif A. M.
Lee, Jae Man
Mon, Hiroaki
Sakashita, Kosuke cc
Li, Mo cc
Kusakabe, Takahiro
Pain, Arnab cc
Hamdan, Samir cc
KAUST Department
Biological and Environmental Sciences and Engineering (BESE) Division
Bioscience Program
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 Number
BAS/1/1020-01-01
Date
2020-08-04
Permanent link to this record
http://hdl.handle.net/10754/664657

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Abstract
One-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.
Citation
Takahashi, 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
Sponsors
This 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.
Publisher
Cold Spring Harbor Laboratory
DOI
10.1101/2020.07.31.20165704
Additional Links
http://medrxiv.org/lookup/doi/10.1101/2020.07.31.20165704
Collections
Biological and Environmental Science and Engineering (BESE) Division; Preprints; Bioscience Program