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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.
  • Mascuch SJ; School of Biological Sciences, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Fakhretaha-Aval S; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Bowman JC; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA; Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Ma MTH; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Thomas G; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Bommarius B; Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, Georgia, USA; School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Ito C; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Zhao L; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA; Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Newnam GP; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Matange KR; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Thapa HR; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Barlow B; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Donegan RK; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Nguyen NA; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Saccuzzo EG; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Obianyor CT; School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Karunakaran SC; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Pollet P; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Rothschild-Mancinelli B; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Mestre-Fos S; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Guth-Metzler R; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Bryksin AV; Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Petrov AS; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Hazell M; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Ibberson CB; School of Biological Sciences, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Penev PI; School of Biological Sciences, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Mannino RG; Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, Georgia, USA.
  • Lam WA; Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, Georgia, USA; Institute for Electronics and Nanotechnology, Georgia Institute of Technology, Atlanta, Georgia, USA; Aflac Cancer and Blood Disorders Center, Children's Healthcare o
  • Garcia AJ; Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, Georgia, USA; School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Kubanek J; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA; Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Agarwal V; School of Biological Sciences, Georgia Institute of Technology, Atlanta, Georgia, USA; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA; Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Hud NV; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA; Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Glass JB; Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, Georgia, USA; School of Earth and Atmospheric Sciences, Georgia Institute of Technology, Atlanta, Georgia, USA. Electronic address: covid-test-support@gatech.edu.
  • Williams LD; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA; Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, Georgia, USA.
  • Lieberman RL; School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia, USA; Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, Georgia, USA.
J Biol Chem ; 295(46): 15438-15453, 2020 11 13.
Article in English | MEDLINE | ID: covidwho-975108
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.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Reagent Kits, Diagnostic / Universities / Technology Transfer / COVID-19 Nucleic Acid Testing / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study Limits: Humans Language: English Journal: J Biol Chem Year: 2020 Document Type: Article Affiliation country: Jbc.RA120.015434

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Reagent Kits, Diagnostic / Universities / Technology Transfer / COVID-19 Nucleic Acid Testing / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study Limits: Humans Language: English Journal: J Biol Chem Year: 2020 Document Type: Article Affiliation country: Jbc.RA120.015434