COVID-19: Enhancing testing and sequencing capacity for Severe Acute Respiratory Syndrome Coronavirus 2 and other zoonotic viruses at the Minnesota Veterinary Diagnostic Laboratory

项目摘要/摘要 提高严重急性呼吸综合征冠状病毒2型检测和测序能力 以及明尼苏达州兽医诊断实验室的其他人畜共患病病毒 明尼苏达州兽医诊断实验室(MVDL)有能力检测SARS-CoV-2 用聚合酶链式反应。MVDL在应对大流行的第一阶段通过测试人类 与此同时，明尼苏达州已具备应对检测需求的能力。MVDL是 明尼苏达州动物样本中SARS-CoV-2检测的参考实验室。本建议书要求 为提高实验室通过聚合酶链式反应检测SARS-CoV-2和测序能力的设备提供资金 阳性样本。这些改进还将有利于应对其他人畜共患病的能力 疾病。SARS-CoV-2聚合酶链式反应测试目前在应用生物系统7500热系统中进行 Cycler，一次可以测试93个样本。为提高程序的效率，我们建议 购买一台QuantStudio 6 Pro热循环仪，一次可以处理378个样品。按顺序排列 SARS-Co-2阳性标本是疾病监测的重要组成部分。排序允许 确定新的变种并监测令人关注的变种。MVDL执行例程 对多种病毒病原体进行测序，如果需要，可以轻松开始对SARS-CoV-2进行测序。这个 MVDL拥有从各种样本中对病毒进行测序的专业知识，其中一些样本的病毒含量很低 DNA/RNA。作为测序过程的一部分执行的初始PCR扩增步骤目前 使用ABI 9700 GenAmp聚合酶链式反应仪器，这些仪器很旧，不需要维护和维修 零件不再有货了。因此，我们建议从生活购买一台普瑞克斯96孔热循环仪 科技/Thermo Fisher。这台仪器的增加将增加实验室的测序能力 除了目前的常规测序程序外，还可以进行SARS-CoV-2测序 正在奔跑。

Project Summary/Abstract Enhancing testing and sequencing capacity for Severe Acute Respiratory Syndrome Coronavirus 2 and other zoonotic viruses at the Minnesota Veterinary Diagnostic Laboratory The Minnesota Veterinary Diagnostic Laboratory (MVDL) has the capability of testing for SARS-CoV-2 by PCR. The MVDL was critical during the first phase of the response to the pandemic by testing human samples while the state of Minnesota acquired the capacity to respond to the testing needs. The MVDL is the reference laboratory for SARS-CoV-2 testing in animal samples in Minnesota. This proposal requests funding for equipment to improve laboratory capacity to test for SARS-CoV-2 by PCR and to sequence positive samples. These improvements would also benefit the capacity to respond to other zoonotic diseases. The SARS-CoV-2 PCR test is currently performed in an Applied Biosystems 7500 thermal cycler, which can test 93 samples at a time. To improve the efficiency of the process, we propose to purchase a Quantstudio 6 Pro thermal cycler, which can process 378 samples at a time. Sequencing of SARS-Co-2-positive samples is an important component of disease surveillance. Sequencing allows for identification of new variants and monitoring of variants of concern. The MVDL performs routine sequencing for multiple viral pathogens, and could easily start sequencing SARS-CoV-2 if needed. The MVDL has expertise sequencing viruses from a wide variety of specimens, some with low amounts of DNA/RNA. The initial PCR amplification step performed as part of the sequencing process is currently done with ABI 9700 GenAmp PCR instruments, which are old and do not have maintenance and repair parts available anymore. Therefore, we propose to purchase a Proflex 96-well thermal cycler from Life Technologies/Thermo Fisher. The addition of this instrument will add sequencing capacity to the lab so SARS-CoV-2 sequencing can be performed in addition to the routine sequencing procedures currently being run.