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

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 su...

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Bibliographic Details
Published in:	The Journal of biological chemistry Vol. 295; no. 46; pp. 15438 - 15453
Main Authors:	Mascuch, Samantha J., Fakhretaha-Aval, Sara, Bowman, Jessica C., Ma, Minh Thu H., Thomas, Gwendell, Bommarius, Bettina, Ito, Chieri, Zhao, Liangjun, Newnam, Gary P., Matange, Kavita R., Thapa, Hem R., Barlow, Brett, Donegan, Rebecca K., Nguyen, Nguyet A., Saccuzzo, Emily G., Obianyor, Chiamaka T., Karunakaran, Suneesh C., Pollet, Pamela, Rothschild-Mancinelli, Brooke, Mestre-Fos, Santi, Guth-Metzler, Rebecca, Bryksin, Anton V., Petrov, Anton S., Hazell, Mallory, Ibberson, Carolyn B., Penev, Petar I., Mannino, Robert G., Lam, Wilbur A., Garcia, Andrés J., Kubanek, Julia, Agarwal, Vinayak, Hud, Nicholas V., Glass, Jennifer B., Williams, Loren Dean, Lieberman, Raquel L.
Format:	Journal Article
Language:	English
Published:	United States Elsevier Inc 13-11-2020 American Society for Biochemistry and Molecular Biology
Subjects:	Biotechnology - methods coronavirus COVID-19 - diagnosis COVID-19 - virology COVID-19 Nucleic Acid Testing - methods DNA polymerase Editors' Picks formulation Humans infectious disease polymerase chain reaction (PCR) protein purification Reagent Kits, Diagnostic - economics Reagent Kits, Diagnostic - supply & distribution Real-Time Polymerase Chain Reaction - methods reverse transcriptase reverse transcription ribonuclease inhibitor RNA RT-qPCR SARS-CoV-2 SARS-CoV-2 - genetics SARS-CoV-2 - isolation & purification Technology Transfer Universities - economics virus DNA polymerase RNA reverse transcriptase protein purification coronavirus virus reverse transcription ribonuclease inhibitor SARS-CoV-2 infectious disease formulation polymerase chain reaction (PCR) RT-qPCR
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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. 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.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Edited by Craig E. Cameron These authors contributed equally to this work.
ISSN:	0021-9258 1083-351X
DOI:	10.1074/jbc.RA120.015434