Novel Approaches to detect and treat sepsis

项目摘要/摘要： 背景资料。败血症是一种严重的危及生命的疾病，是中国最常见的死亡原因之一 住院病人。败血症通常是由细菌感染引起的，包括革兰氏阴性和 阳性细菌。在美国，脓毒症患者的住院死亡率可能高达 41.1%，每年造成超过25万人死亡和200亿美元的损失。由于资金不足， 由于目前技术的敏感性和特异性，脓毒症的诊断尚无全球标准。在这 作为该项目的一部分，PI有雄心解决败血症检测中特别令人担忧的关键瓶颈问题，方法是： 1)混合生物-无机纳米机器人，2)基于CRISPR的设备，以及3)配备CRISPR的工程噬菌体。 未来五年的目标。我们的第一个目标是在衣壳蛋白上设计具有纳米抗体的噬菌体M13 尾部纤维蛋白PIII上的pviii和His-tag。在结合了镀钴的磁性纳米颗粒后，所产生的 混合生物-无机纳米机器人将用于浓缩和净化血液样本中的病原体。俘获 将使用添加的样本来研究效率，然后进行临床试验。利用 CRISPR和微流控技术，第二个目标是制造便携式设备来检测与脓毒症相关的 病原体，可以在资源有限的情况下使用。最后一个目标是用不同的基因工程噬菌体 CRISPR系统，可用于检测和抗击败血症相关的细菌病原体。接近尾声的时候 在第五年，我们将整合这些技术，作为一个强大的脓毒症诊断工具。 研究计划的总体构想。我们正在开发的技术将对 生物医学研究社区发现和治疗脓毒症，甚至是其他疾病。我们开发的 技术还可以促进其他领域的病原体检测，如食品安全和环境 监控。

Project Summary/Abstract: Background. Sepsis, a severe and life-threatening condition, is one of the most common causes of death in hospitalized patients. Sepsis is generally caused by bacterial infection, including both Gram-negative and positive bacteria. In the United States, the hospital mortality rate of patients with sepsis could be as high as 41.1%, which accounts for more than 250,000 deaths and $20 billion loss annually. Due to the inadequate sensitivity and specificity of the current technologies, there is no global standard for sepsis diagnosis. In this project, the PI has the ambition to address the critical bottlenecks specifically of concern in sepsis testing using: 1) hybrid bio-inorganic nanobots, 2) CRISPR-based devices, and 3) CRISPR-equipped engineered phages. Goals for the next five years. Our first goal is to engineer phage M13 with nanobodies on the capsid protein pVIII and his-tags on the tail fiber protein pIII. After binding cobalt-coated magnetic nanoparticles, the resulting hybrid bio-inorganic nanobots will be used to concentrate and purify pathogens from blood samples. Capture efficiency will be investigated using spiked samples and then proceed to clinical ones. Taking advantage of CRISPR and microfluidic technologies, the second goal is to fabricate portable devices to detect sepsis-related pathogens, which can be used in resource-limited settings. The last goal is to engineer phages with different CRISPR systems, that can be used to detect and combat sepsis-related bacterial pathogens. Towards the end of the fifth year, we will have integrated these technologies as a robust tool for sepsis diagnosis. Overall vision of the research program. The technologies we are developing will have a broad impact on the biomedical research communities to detect and treat sepsis, even for other diseases. Our developed technologies can also advance pathogen detection in other fields, such as food safety and environmental monitoring.