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

项目总结/摘要 加强严重急性呼吸系统综合症冠状病毒2型的检测和测序能力 和其他人畜共患病毒在明尼苏达州兽医诊断实验室 明尼苏达州兽医诊断实验室（MVDL）有能力检测SARS-CoV-2 用PCR MVDL在应对大流行的第一阶段至关重要， 样本，而明尼苏达州获得的能力，以应对测试的需要。MVDL是 在明尼苏达州的动物样本中进行SARS-CoV-2测试的参考实验室。该提案要求 提供资金，用于提高实验室通过聚合酶链反应检测SARS-CoV-2和测序的能力 阳性样本。这些改进也将有利于应对其他人畜共患病的能力。 疾病SARS-CoV-2 PCR检测目前在Applied Biosystems 7500热分析仪中进行。 循环仪，一次可以测试93个样本。为提高程序的效率，我们建议 购买Quantstudio 6 Pro热循环仪，可一次处理378个样本。测序 SARS-Co-2阳性样本是疾病监测的重要组成部分。测序允许 识别新的变体并监测相关变体。MVDL执行例程 测序多种病毒病原体，并可以很容易地开始测序SARS-CoV-2，如果需要的话。的 MVDL拥有对来自各种标本的病毒进行测序的专业知识，其中一些标本的病毒含量较低， DNA/RNA。作为测序过程的一部分进行的初始PCR扩增步骤目前是 使用ABI 9700 GenAmp PCR仪器完成，该仪器较旧且无法维护和维修 零件不再可用。因此，我们建议从Life购买Proflex 96孔热循环仪 技术/Thermo Fisher。该仪器的增加将增加实验室的测序能力， SARS-CoV-2测序可以在目前常规测序程序之外进行 正在运行。

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.