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

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.

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.