PDF(Pubmed)
Abstract：
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.
摘要：
在有效治疗之前，在个体中广泛检测新型冠状病毒SARS-CoV-2的存在对于控制COVID-19大流行仍然至关重要。测试中的挑战可以追溯到最初的供应短缺，通过定量逆转录聚合酶链反应（RT-qPCR）检测病毒所需的专业知识和/或仪器，最健壮的，敏感,和目前可用的特定测定。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-2RT-qPCR测试试剂盒和回填管道短缺。佐治亚理工学院COVID-19测试套件支持小组，由教师组成，工作人员,佐治亚理工学院生物技术学院的学员，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。我们还开发了一种环境测试协议，可以轻松监控各个校园实验室中是否存在SARS-CoV-2的表面。我们的蓝图应该很容易被其他机构的研究团队复制，我们的协议可能会被修改和调整，以便在更多资源有限的环境中检测SARS-CoV-2。
