A blueprint for academic labs to produce SARS-CoV-2 RT-qPCR test kits

• Authors: Samantha J. Mascuch, Georgia Institute of Technology; Sara Fakhretaha-Aval, Georgia Institute of Technology; Jessica C. Bowman, Georgia Institute of Technology; Minh Thu H. Ma, Georgia Institute of Technology; Gwendell Thomas, Georgia Institute of Technology; Bettina Bommarius, Georgia Institute of Technology; Chieri Ito, Georgia Institute of Technology; Liangjun Zhao, Georgia Institute of Technology; Gary P. Newnam, Georgia Institute of Technology; Kavita R. Matange, Georgia Institute of Technology; Hem R. Thapa, Georgia Institute of Technology; Brett Barlow, Georgia Institute of Technology; Rebecca K. Donegan, Georgia Institute of Technology; Nguyet A. Nguyen, Georgia Institute of Technology; Emily G. Saccuzzo, Georgia Institute of Technology; Chiamaka T. Obianyor, Georgia Institute of Technology; Suneesh C. Karunakaran, Georgia Institute of Technology; Pamela Pollet, Georgia Institute of Technology; Brooke Rothschild-Mancinelli, Georgia Institute of Technology; Santi Mestre-Fos, Georgia Institute of Technology; Rebecca Guth-Metzler, Georgia Institute of Technology; Anton V. Bryksin, Georgia Institute of Technology; Anton S. Petrov, Georgia Institute of Technology; Mallory Hazell, Georgia Institute of Technology; Carolyn B. Ibberson, Georgia Institute of Technology; Petar I. Penev, Georgia Institute of Technology; Robert Mannino, Emory University; Wilbur Lam, Emory University; Andrés J. Garcia, Georgia Institute of Technology; Julia M. Kubanek, Georgia Institute of Technology; Vinayak Agarwal, Georgia Institute of Technology; Nicholas V. Hud, Georgia Institute of Technology; Jennifer B. Glass, Georgia Institute of Technology; Loren Dean Williams, Georgia Institute of Technology; Raquel L. Liberman, Georgia Institute of Technology
• Journal Title: medRxiv
• Volume: Volume 2
• Publisher: NIH | 2020-09-01
• Type of Work: Article | Preprint: Prior to Peer Review
• Publisher DOI: http://doi.org/10.1101/2020.07.29.20163949
• Subjects: Biology, Virology; Health Sciences, Health Care Management
• Keywords: reverse transcriptase; polymerase chain reaction; ribonuclease inhibitor; DNA polymerase; virus; formulation; SARS-CoV-2

Abstract

Widespread testing for the presence of the novel coronavirus 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 transcription polymerase chain reaction (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 to a commercial product used for diagnostic testing. We also developed an environmental testing protocol to readily monitor surfaces across various campus laboratories 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.