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
School of Biological Sciences, Georgia Institute of Technology, Atlanta, GA, USA.
School of Chemistry & Biochemistry, Georgia Institute of Technology, Atlanta, GA, USA.
medRxiv. 2020 Sep 1:2020.07.29.20163949. doi: 10.1101/2020.07.29.20163949.
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.
在有效治疗方法出现之前，对个体进行新型冠状病毒SARS-CoV-2检测对于控制COVID-19大流行仍然至关重要。检测方面的挑战可追溯到最初缺乏通过定量逆转录聚合酶链反应（RT-qPCR）检测病毒所需的物资、专业知识和/或仪器，RT-qPCR是目前可用的最可靠、灵敏和特异的检测方法。在此，我们表明配备适当专业知识和基础设施的学术生物化学与分子生物学实验室能够复制市售的SARS-CoV-2 RT-qPCR检测试剂盒，并填补检测流程中的短缺。佐治亚理工学院COVID-19检测试剂盒支持小组由佐治亚理工学院生物技术学院的教师、工作人员和学员组成，他们合成了多重引物和探针，并配制了一种由内部生产的酶和蛋白质组成的预混液。我们的内部试剂盒与用于诊断检测的商业产品相比毫不逊色。我们还制定了一项环境检测方案，以便随时监测各个校园实验室的表面是否存在SARS-CoV-2。我们的方案蓝图应该很容易被其他机构的研究团队复制，并且我们的方案可以修改和调整，以便在资源更有限的环境中进行SARS-CoV-2检测。
