Isothermal Chain Reaction (ICR) Rapid and Early Detection of Pathogens for Sepsis Pont-of-Care Testing, Stratification, and Monitoring

项目摘要 目标： 该应用程序长期目标是开发用于常规护理点(POC)遗传的新产品 测试，范围从病原体识别到早期癌症检测，使用创新的生物技术 称为等温链反应(ICR)。ICR是一个颠覆性的支持技术平台，它将提供 与生物医学研究和医疗保健相关的新型试剂。ICR不需要任何 酶，引子，核苷酸，主混合物，延伸反应，cDNA中间体，或热循环。 这些好处克服了基于聚合酶链式反应的技术的限制，因为它们对发现的酶抑制剂具有抵抗力。 在实验室标本和临床标本中，对罕见突变具有较高的特异性，并降低了试剂 成本。ICR有望实现选择性、快速和负担得起的生物标记物检测，而无需投资 在昂贵的仪器和劳动密集型的样品制备程序中。设想的产品将是 用于在非实验室环境中早期检测传染病和遗传病的分析、试板和试剂盒。 具体目标： 1.脓毒症靶序列ICR探针的开发：我们将设计和开发ICR检测方法，以快速 并在混合种群中等温检测与脓毒症相关的合成靶序列 模板在等温条件下不到10分钟即可完成，无需酶、引物、核苷酸或 大师级调音师。 2.血液样本中已知的脓毒症病原体的鉴定：我们将从特定的目标1到 从回溯血样中确定已知的败血症病原体。ICR结果将作为基准 血培养以证明使用ICR快速早期检测脓毒症生物标志物的可行性。 3.血液样本中未知脓毒症病原体的鉴定：我们将从特定的目标1使用ICR检测 从回顾性血样中鉴定未知的脓毒症病原体。ICR结果将是 以血培养为基准，证明ICR增强了检测选择性的可行性 血培养方法遗漏了病原体，ICR更容易进行护理点败血症检测。 影响： 完成特定目标可能会显著影响败血症病原体的快速和早期检测 当治疗干预措施更有效，并使常规临床测试和筛查成为可能时 以改善临床分诊决策、常规监测和治疗结果。

Project Summary Objectives: The long-term objective of this application is to develop novel products for routine point-of-care (POC) genetic testing, ranging from pathogen identification to early cancer detection by using an innovative biotechnology called, Isothermal Chain Reaction (ICR). ICR is a disruptive and enabling technology platform that will offer a new class of reagents relevant to biomedical research and medical healthcare. ICR does not require any enzymes, primers, nucleotides, master mixes, extension reactions, cDNA intermediates, or thermocycling. These benefits overcome constraints of PCR-based technologies by being resistant to enzyme inhibitors found in laboratory samples and clinical specimens, having higher specificity for rare mutations, and lowering reagent costs. ICR is anticipated to enable selective, rapid, and affordable biomarker detection without having to invest in costly instruments and labor-intensive sample preparation procedures. The envisioned products will be assays, panels, and kits for the early detection of infectious and genetic diseases in non-laboratory settings. Specific Aims: 1. Development of ICR probes to sepsis target sequences: We will design and develop ICR assays to rapidly and isothermally detect synthetic target sequences associated with sepsis in a mixed population of templates in less than 10 minutes under isothermal conditions, without enzymes, primers, nucleotides, or master mixes. 2. Identification of known sepsis pathogens in blood samples: We will use ICR assays from Specific Aim 1 to identify known sepsis pathogens from retrospective blood samples. ICR results will be benchmarked to blood cultures to demonstrate the feasibility of using ICR for rapid early detection of sepsis biomarkers. 3. Identification of unknown sepsis pathogens in blood samples: We will use ICR assays from Specific Aim 1 to identify unknown sepsis pathogens from retrospective blood samples. ICR results will be benchmarked to blood cultures to demonstrate the feasibility that ICR has enhanced selectivity to detect pathogens missed by blood culture methods, and ICR is more amenable to point-of-care sepsis testing. Impact: Accomplishing the Specific Aims could significantly impact rapid and early detection of sepsis pathogens when therapeutic interventions are much more effective, and enable routine clinical testing and screening of the disease to improve clinical triage decisions, routine monitoring, and treatment outcomes.