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.
Highlights
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 reverse transcriptase (RT)-PCR (RT-qPCR), the most robust, sensitive, and specific assay currently available
We focused on producing the N1 and N2 primer and probe system published by Centers for Disease Control (CDC) in March of 2020 (Table 1) because (i) these sequences had been extensively verified in the literature, (ii) our own bioinformatics analysis showed them to be highly specific to SARS-CoV-2 and localized to regions of the genome with low mutation rates, and (iii) they had received Food and Drug Administration (FDA) Emergency Use Authorization (EUA)
Summary
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. 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. Countries to be effective in monitoring and managing the outbreak These strategies will continue to be instrumental in containing the virus until a vaccine or other effective treatment is universally available [4]. The Georgia Tech COVID-19 Test Kit Support Group was conceived to leverage in-house Georgia Tech facilities, expertise, and personnel to assist the State of Georgia Clinical Laboratory Improvement Amendments (CLIA) laboratory with materials needed for clinical SARS-CoV-2 detection. Ours is the first effort to use all in-house materials and equipment, offering an openaccess community resource for other settings with similar capabilities
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