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    Quick and Easy Assembly of a One-step qRT-PCR Kit for COVID-19 Diagnostics Using In-House Enzymes

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    smarthealth2021.mkv2334.NORMAL.pdf
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    Type
    Poster
    Authors
    Takahashi, Masateru
    Tehseen, Muhammad cc
    Salunke, Rahul
    Takahashi, Etsuko
    Mfarrej, Sara
    Sobhy, Mohamed A.
    Alhamlan, Fatimah S.
    Hala, Sharif
    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
    Date
    2021-11-22
    Permanent link to this record
    http://hdl.handle.net/10754/673410
    
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    Abstract
    One-step 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 the 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 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 qRT-PCR and TaqPath 1-Step RT-qPCR kits. Overall, our kit has shown robust performance in both laboratory settings and the Saudi Ministry of Health-approved testing facility.
    Conference/Event name
    KAUST R3T Week 2021
    Collections
    KAUST R3T Week 2021; Posters

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