Journal
JOURNAL OF BIOLOGICAL CHEMISTRY
Volume 295, Issue 46, Pages 15438-15453Publisher
ELSEVIER
DOI: 10.1074/jbc.RA120.015434
Keywords
reverse transcriptase; ribonuclease inhibitor; DNA polymerase; virus; formulation; SARS-CoV-2; reverse transcription; polymerase chain reaction (PCR); infectious disease; protein purification; RNA; coronavirus; RT-qPCR
Categories
Funding
- State of Georgia COVID-19 Testing Task Force Method Development and Supply Chain Stabilization Studies Proposal (COVID-19 Tech Support Group)
- Georgia Institute of Technology
- NASA [80NSSC18K1139, 80NSSC19K0477]
- National Institutes of Health [U54EB027690]
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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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