A blueprint for academic laboratories to produce SARS-CoV-2 quantitative RT-PCR test kits.
A blueprint for academic laboratories to produce SARS-CoV-2 quantitative RT-PCR test kits.
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DOI：
10.1074/jbc.ra120.015434
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发表时间：
2020-11-13
期刊：
影响因子：
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通讯作者：
Lieberman RL
中科院分区：
文献类型：
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作者：
Mascuch SJ;Fakhretaha-Aval S;Bowman JC;Ma MTH;Thomas G;Bommarius B;Ito C;Zhao L;Newnam GP;Matange KR;Thapa HR;Barlow B;Donegan RK;Nguyen NA;Saccuzzo EG;Obianyor CT;Karunakaran SC;Pollet P;Rothschild-Mancinelli B;Mestre-Fos S;Guth-Metzler R;Bryksin AV;Petrov AS;Hazell M;Ibberson CB;Penev PI;Mannino RG;Lam WA;Garcia AJ;Kubanek J;Agarwal V;Hud NV;Glass JB;Williams LD;Lieberman RL
在有效治疗方法出现之前，在个体中广泛检测新型冠状病毒严重急性呼吸综合征冠状病毒2 （SARS-CoV-2）的存在对于控制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RT-PCR （RT-qPCR）检测病毒所需的供应、专业知识和/或仪器。RT-qPCR是目前可用的最强大、最敏感和最特异的检测方法。在这里，我们表明，配备适当专业知识和基础设施的学术生物化学和分子生物学实验室可以复制市售的SARS-CoV-2 RT-qPCR检测试剂盒，并填补管道短缺。佐治亚理工学院COVID-19测试试剂盒支持小组由佐治亚理工学院生物技术学院的教职员工和实习生组成，他们合成了多重引物和探针，并配制了由内部生产的酶和蛋白质组成的主混合物。我们的内部套件与用于诊断测试的商业产品相比具有优势。我们还制定了一项环境测试方案，以便随时监测表面是否存在SARS-CoV-2。我们的蓝图应该很容易被其他机构的研究团队复制，我们的方案可能会被修改和调整，以便在资源更有限的环境中检测到SARS-CoV-2。
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.