Endotoxin Quantification in Aqueous Media

描述(由申请人提供)：在美国，每年大约有75万例败血症导致20万人死亡[1]。由于很难对抗内毒素的影响，许多医学专家建议，预防内毒素引起的脓毒症的最好方法是严格的内毒素筛选过程[2，3]。拟议的研究旨在开发一种基于一次性远程查询磁弹性传感器的低成本、因此可广泛使用的、高灵敏度和快速的内毒素检测系统。该系统利用磁弹性传感器平台的黏度监测功能，通过测量凝胶形成法(LAL)中的凝胶形成来确定内毒素水平，LAL法是一种流行的从马蹄蟹中提取的内毒素定量方法。该传感器的微型特性，加上其出色的粘度灵敏度，使微升分析能够显著降低成本并便于使用。我们相信，拟议的系统将使医疗保健提供者能够轻松、快速、廉价地检测高危患者的内毒素感染，从而显著降低脓毒症死亡率。浸泡在LAL化验溶液中的无源带状磁弹性传感器可通过磁场远程检测。初步研究表明，在LAL测定的凝血过程中，磁弹性传感器的共振幅度减小，时间变化率的大小与内毒素浓度成正比。这项工作的重点是进行使能的科学和工程，将促进磁弹性内毒素检测设备的开发和优化其性能。具体地说，在两年的项目期限内，我们将：{1}进一步微型化和优化传感器，以减少所需的LAL化验量并提高灵敏度。{2}改进所需的传感器读取器电子设备，扩展频率操作上限，并包括额外的40分贝增益。{3}开发一种技术和/或算法，根据传感器的初始响应准确计算内毒素水平，从而在几分钟内实现内毒素定量。{4}设计并制作了传感器和样品采集器的无热原拉尔胶一次性包装。我们寻求通过为医疗保健提供者提供一种准确、廉价和快速的工具来检测高危患者的内毒素感染，从而显著降低脓毒症死亡率。考虑到每年因脓毒症导致的死亡率，超过20万人死亡，我们相信这个项目的成功将为改善公共卫生做出重大贡献。

DESCRIPTION (provided by applicant): Annually in the United States there are approximately 750,000 cases of sepsis resulting in 200,000 deaths [1]. Since it is difficult to counter the effects of endotoxin, many medical experts suggest that the best way to prevent endotoxin-induced sepsis is a rigorous endotoxin screening process [2,3]. The proposed research seeks to enable development of a low-cost, hence widely available, highly sensitive and rapid endotoxin detection system based on the use of disposable remote-query magnetoelastic sensors. Utilizing the viscosity-monitoring capabilities of the magnetoelastic sensor platform, this system determines the endotoxin level by measuring the gel formation in Limulus Amoebocyte Lysate (LAL) assay, a popular assay derived from horseshoe crabs for endotoxin quantification. The miniature nature of the sensor, coupled with its excellent viscosity sensitivity, enables micro-Liter assays significantly reducing cost and facilitating ease of use. We believe that the proposed system will significantly reduce the rate of sepsis mortality by allowing healthcare providers to easily, rapidly, and inexpensively detect endotoxic infections in high-risk patients. Passive, ribbon-like magnetoelastic sensors, immersed within the LAL assay-test solution, are remotely interrogated via magnetic fields. Preliminary studies have shown that the resonance amplitude of the magnetoelastic sensor decreases during the coagulation of the LAL assay, and the magnitude of the time rate of change is proportional to the endotoxin concentration. The focus of this effort is to conduct the enabling science and engineering that will facilitate development of the magnetoelastic endotoxin detection device and optimize its performance. Specifically, over the two year project duration we will: {1} further miniaturize and optimize the sensors to reduce the amount of LAL assay needed with enhanced sensitivity. {2} refine the needed sensor reader electronics, extending the upper frequency operational limit and including an additional 40dB of gain. {3} develop a technique and/or algorithm to accurately calculate the endotoxin level based on the initial response of the sensor to enable endotoxin quantification in but a few minutes time. {4} Design and fabricate the pyrogen-free LAL-mixing disposable package for the sensor and the sample collection apparatus. We seek to significantly reduce the rate of sepsis mortality by providing healthcare providers an accurate, inexpensive, and rapid tool for detecting endotoxic infections in at-risk patients. Considering the annual mortality rate due to sepsis, over 200,000 deaths, we believe the success of this project will make a significant contribution in improving public health.