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author 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 M.
Agarwal, Vinayak
Hud, Nicholas V.
Glass, Jennifer B.
Williams, Loren Dean
Lieberman, Raquel L.
author_facet 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 M.
Agarwal, Vinayak
Hud, Nicholas V.
Glass, Jennifer B.
Williams, Loren Dean
Lieberman, Raquel L.
author_sort Mascuch, Samantha J.
collection PubMed
description 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.
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spelling pubmed-74020632020-08-06 A blueprint for academic labs to produce SARS-CoV-2 RT-qPCR 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 M. Agarwal, Vinayak Hud, Nicholas V. Glass, Jennifer B. Williams, Loren Dean Lieberman, Raquel L. medRxiv Article 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. Cold Spring Harbor Laboratory 2020-09-01 /pmc/articles/PMC7402063/ /pubmed/32766604 http://dx.doi.org/10.1101/2020.07.29.20163949 Text en http://creativecommons.org/licenses/by-nc/4.0/It is made available under a CC-BY-NC 4.0 International license (http://creativecommons.org/licenses/by-nc/4.0/) .
spellingShingle Article
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 M.
Agarwal, Vinayak
Hud, Nicholas V.
Glass, Jennifer B.
Williams, Loren Dean
Lieberman, Raquel L.
A blueprint for academic labs to produce SARS-CoV-2 RT-qPCR test kits
title A blueprint for academic labs to produce SARS-CoV-2 RT-qPCR test kits
title_full A blueprint for academic labs to produce SARS-CoV-2 RT-qPCR test kits
title_fullStr A blueprint for academic labs to produce SARS-CoV-2 RT-qPCR test kits
title_full_unstemmed A blueprint for academic labs to produce SARS-CoV-2 RT-qPCR test kits
title_short A blueprint for academic labs to produce SARS-CoV-2 RT-qPCR test kits
title_sort blueprint for academic labs to produce sars-cov-2 rt-qpcr test kits
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7402063/
https://www.ncbi.nlm.nih.gov/pubmed/32766604
http://dx.doi.org/10.1101/2020.07.29.20163949
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