Microarray Assay for Detection of Lab Animal Pathogens

描述(申请人提供)：研究动物在生物医学研究、药物开发和毒理学测试中起着至关重要的作用。据估计，仅在美国就有超过3000万只小鼠和大鼠用于这些目的。未检测到的感染的存在可能危及实验和/或导致有价值的菌株或整个菌落的损失。该项目的目标是开发一种具有成本效益的基于微阵列的血清监测系统，用于对实验动物进行全面的健康评估，特别是小鼠。老鼠监测器TM系统将能够同时检测多达9种相关的老鼠病原体的存在，只需要10-20微升的血清。在第一阶段，对老鼠监测器TM的几个关键部件进行了广泛的评估，包括一个新的专有阵列基板和两个原型平台，证明了这种方法的可行性。在第二阶段，我们的具体目标将集中在：a)重组抗原的表达和验证，b)4病原体监测分析的发展，c)9病原体监测加分析的发展。对于这两个化验小组，将确定和优化灵敏度、特异度和精确度的关键化验参数，以满足性能标准。经过内部验证后，原型老鼠监视器TM系统将被放置在两个合作地点进行测试。Claros诊断公司将为该项目制造一种针对小型实验室的超低成本小说阅读器，该阅读器由一个手持阅读器和相关的预装试剂的微流控墨盒组成。耶鲁大学医学院和动物设施的分子、血清学和病毒学诊断部主任苏珊·康普顿博士将担任顾问并参与Beta测试。杰克逊实验室的诊断服务主管詹姆斯·费希博士也将参与老鼠监测器的贝塔测试，并对其性能提供反馈。杰克逊实验室拥有100多万只小鼠。

DESCRIPTION (provided by applicant): Research animals play a crucial role in biomedical research, drug development and toxicology testing. It is estimated that an average population of over 30 million mice and rats is maintained in the U.S. alone for these purposes. The presence of undetected infections can compromise experiments and/or cause the loss of valuable strains or entire colonies. The goal of this project is to develop a cost-effective microarray-based sero-surveillance system for a comprehensive health assessment of laboratory animals with a special focus on mice. The Murine MonitorTM System will be capable of simultaneously testing for the presence of up to 9 relevant murine pathogens, requiring only 10-20 microliters of serum. During Phase I, several critical components of the Murine MonitorTM have been extensively evaluated including a novel proprietary array substrate and two prototype platforms, demonstrating the feasibility of this approach. During Phase II, our specific aims will focus on: a) expression and validation of recombinant antigens, b) development of a 4-pathogen Surveillance Assay, c) development of a 9-pathogen Surveillance Plus Assay. For both assay panels, the critical assay parameters of sensitivity, specificity and precision will be determined and optimized to meet performance criteria. After internal validation, prototype Murine MonitorTM systems will be placed at two collaborating sites for testing. An ultra-low-cost novel reader aimed at small laboratories consisting of a hand-held reader and associated microfluidic cartridge which is preloaded with reagents will be manufactured for this project by Claros Diagnostics, Inc. Dr. Susan Compton, Chief of the Molecular, Serological and Virological Diagnostics Unit at Yale University School of Medicine and Animal Facility will serve as a consultant and will participate in beta-testing. Dr James Fahey, Chief of Diagnostic Services at The Jackson Laboratory, which houses over 1 million mice, will also participate in beta-testing the Murine Monitor and provide feedback on its performance.