A portable quantitative polymerase chain reaction platform (qPCR) for rapid detection of pathogens impacting model organisms in animal facilities

便携式定量聚合酶链反应平台 (qPCR)，用于快速检测影响动物设施中模式生物的病原体

• 批准号: 10604150
• 负责人: Scott Franklin Geller
• 金额: $ 25.5万
• 依托单位: BAY GENOMICS, LLC
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10604150
• 关键词: Address; Affect; Air; Amphibia; Animal Experimentation; Animal Experiments; Animal Model; Animals; Area; Bacteria; Biological; Biological Assay; Biology; Clinical; Clinical Trials; Collaborations; DNA Sequence; Data; Detection; Development; Devices; Diagnostic; Electronics; Ensure; Experimental Animal Model; Extravasation; Fluorescence; Goals; Graph; Health; Health care facility; Heating; Helicobacter; Hour; Human; Invaded; Laboratory Animals; Lead; Legal patent; Manuals; Marketing; Methods; Microbe; Microfluidics; Monitor; Movement; Mus; Noise; Optics; Outcome; Output; Pathogen detection; Phase; Polymerase Chain Reaction; Population; Preparation; Process; Production; Quarantine; Rapid diagnostics; Reagent; Reproducibility; Research Personnel; Rodent; Running; Sampling; Science; Signal Transduction; Site; Small Business Innovation Research Grant; Specificity; Speed; System; Testing; Time; Translations; Validation; Veterinarians; Virulent; Work; animal care; animal facility; commercialization; design; empowerment; experimental study; fungus; human disease; improved; innovation; innovative technologies; instrument; interest; manufacture; model organism; pathogen; pathogenic microbe; portability; product development; prototype; rapid detection; remediation; response; sample collection; seal; success; usability; wireless electronic

Project Summary In recent years there have been growing concerns about the rigor and reliability of data originating from common and critical animal model experimentation. In fact, a core principal of science – reproducibility – has been called into question largely due to challenges in replicating biologically complicated animal experiments. We have developed a fast, portable, quantitative PCR (the current universal “gold standard” method) device for use in a wide variety of diagnostic settings. We believe that our rapid, handheld, battery-powered and wireless device has the potential to empower rapid diagnostics around the world. In this proposal we aim to validate the use of our prototype device using real-world samples for microbial pathogen detection. Regarding our innovative technology, we have taken a fundamentally different approach to heating and cooling the sample being amplified, avoiding the standard Peltier heating block. The heat transfer is so efficient that we are able to perform a heating and cooling cycle in as little as 15 seconds (possibly even faster), resulting in a 40-cycle run finishing in about 11 minutes. In essence, this means you can get a quantitative answer regarding the presence and abundance of your target pathogen with ultimate sensitivity significantly faster than the current state of the art. Looking deeper into the root causes of animal-derived data variability is pointing to several possible environmental and biological causes. In terms of the biology, microbes that live inside or outside of the body are proving to have clear and present effects on experimental outcomes, yielding data that is inconsistent and questionable. In particular, when the studies of interest are translational by design, and therefore intended to benefit / improve human health, there is even greater cause for concern. We are poised to test our device to identify and quantify the presence (or absence) of pathogens in an fully operating academic animal facility at UC Berkeley. Together with the Office of Laboratory Animal Care, we have identified two important and tractable pathogens as good candidates for initial proof of principle studies: Helicobacter species (bacteria) in mice and B. dendrobatidis, a fungus affecting amphibians.

项目摘要 近年来，人们对来自源自数据的严格性和可靠性越来越担心 常见和关键的动物模型实验。实际上，科学的核心原理 - 可重复性 - 在很大程度上被提出的质疑是由于复制生物学上复杂的动物实验的挑战。 我们已经开发了快速，便携式，定量PCR（当前通用“金标准”方法）设备 用于多种诊断设置。我们相信我们的迅速，手持式，电池供电和 无线设备有可能增强全球快速诊断的能力。在这个建议中，我们的目标是 使用现实世界样品验证使用原型设备进行微生物病原体检测。 关于我们的创新技术，我们采取了一种根本不同的方法来供暖和 冷却样品被放大，避免标准的毛发加热块。传热是如此 高效，我们能够在短短15秒内执行加热和冷却周期（甚至可能 更快），在大约11分钟内完成40周期的跑步。 本质上，这意味着您可以获得有关您的存在和抽象的定量答案 靶病原体具有最终灵敏度的速度明显快于当前的技术状态。 深入研究动物来源数据可变性的根本原因指向了几种可能的 环境和生物学原因。就生物学而言，生活在体内或体外的微生物 事实证明，对实验结果产生明确的影响，产生不一致的数据，并且 值得怀疑的。特别是，当感兴趣的研究按设计翻译时，因此旨在 受益 /改善人类健康，还有更大的关注理由。 我们被中毒以测试我们的设备，以识别和量化完全的病原体的存在（或不存在） 在UC Berkeley运营学术动物设施。与实验室动物护理办公室一起，我们 已经确定了两种重要且可进行的病原体是原则研究初始证明的良好候选者： 小鼠和树突状芽孢杆菌（一种影响两栖动物的真菌）中的螺旋杆菌物种（细菌）。