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，越来越多的人关注数据的严谨性和可靠性， 常见和关键的动物模型实验。事实上，科学的一个核心原则--再现性-- 在很大程度上，由于在复制生物学上复杂的动物实验方面的挑战，人们对此提出了质疑。 我们研制了一种快速、便携、定量PCR（目前通用的“金标准”方法）装置 用于各种各样的诊断设置。我们相信，我们的快速，手持，电池供电， 无线设备有可能使世界各地的快速诊断成为可能。在本建议中，我们的目标是 使用真实样本验证我们的原型设备用于微生物病原体检测。 关于我们的创新技术，我们采取了一种完全不同的加热方法， 冷却被扩增的样品，避免标准的Peltier加热块。热传递是如此 我们能够在短至15秒（甚至可能是15秒）内执行加热和冷却循环。 更快），导致40个循环运行在约11分钟内完成。 从本质上讲，这意味着你可以得到一个关于你的存在和丰富的定量答案。 目标病原体的最终灵敏度显著快于现有技术水平。 深入研究动物源性数据变异性的根本原因， 环境和生物原因。从生物学的角度来说，生活在体内或体外的微生物 被证明对实验结果有明确和现实的影响，产生的数据是不一致的， 值得怀疑特别是，当感兴趣的研究通过设计是平移的，因此旨在 虽然这对人类健康有益/改善，但更令人关切的是。 我们准备测试我们的设备，以确定和量化病原体的存在（或不存在）， 经营加州大学伯克利分校的学术动物设施与实验动物护理办公室一起，我们 已经确定了两种重要且易于处理的病原体作为初步原理验证研究的良好候选物： 小鼠和B.一种影响两栖动物的真菌。