A blueprint for academic laboratories to produce SARS-CoV-2 quantitative RT-PCR test kits

学术实验室生产 SARS-CoV-2 定量 RT-PCR 检测试剂盒的蓝图

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作者:Samantha J Mascuch, Sara Fakhretaha-Aval, Jessica C Bowman, Minh Thu H Ma, Gwendell Thomas, Bettina Bommarius, Chieri Ito, Liangjun Zhao, Gary P Newnam, Kavita R Matange, Hem R Thapa, Brett Barlow, Rebecca K Donegan, Nguyet A Nguyen, Emily G Saccuzzo, Chiamaka T Obianyor, Suneesh C Karunakaran, Pame

Abstract

Widespread testing for the presence of the novel coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in individuals remains vital for controlling the COVID-19 pandemic prior to the advent of an effective treatment. Challenges in testing can be traced to an initial shortage of supplies, expertise, and/or instrumentation necessary to detect the virus by quantitative RT-PCR (RT-qPCR), the most robust, sensitive, and specific assay currently available. Here we show that academic biochemistry and molecular biology laboratories equipped with appropriate expertise and infrastructure can replicate commercially available SARS-CoV-2 RT-qPCR test kits and backfill pipeline shortages. The Georgia Tech COVID-19 Test Kit Support Group, composed of faculty, staff, and trainees across the biotechnology quad at Georgia Institute of Technology, synthesized multiplexed primers and probes and formulated a master mix composed of enzymes and proteins produced in-house. Our in-house kit compares favorably with a commercial product used for diagnostic testing. We also developed an environmental testing protocol to readily monitor surfaces for the presence of SARS-CoV-2. Our blueprint should be readily reproducible by research teams at other institutions, and our protocols may be modified and adapted to enable SARS-CoV-2 detection in more resource-limited settings.

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