Targeted Biomarker Panels and Pre-processing Device for the Rapid Assessment of Radiation Injury in Easily Accessible Biofluids

这项建议建立在我们在非靶向和靶向代谢组学方面的专业知识基础上 产生高灵敏度和定量的靶向多重分析。产生这样一个概念背后的想法 在容易获得的生物体液(尿液、血液、唾液)中进行辐射评估和辐射损伤检测的方法是 迅速确定个人接触的程度，并区分担心的健康和 可能需要医疗干预的接触者。将采取高度量化的方法 通过液-质联用(LC-MS/MS)对已鉴定的每一个 每种生物流体中的辐射生物标记物。这种仪器目前在临床实验室中被常规使用， 因此，最大限度地利用可用资源，在 紧急情况。基于灵敏度、高信噪比、矩阵低信号抑制标准 作用，以及与对照或代谢产物对之间的关系相比的高倍数变化， 将收集生物签名并计算浓度。将开发组合的生物签名 在多重分析中，有效地减少了从样品准备到结果的时间。我们的目标是 证明这种多重分析方法具有在紧急情况下部署的潜力 预先确定的临床实验室网络，可以接受并快速处理大量 样本。虽然最终目标将是让这样一个小组在所有情况下都具有预测性，即使是有限的错误 在大规模伤亡的情况下，阳性率将有助于评估辐射损伤：例如，错误1% 阳性率将使需要进一步评估的人数减少100倍。此外，这一点 检测将是灵活的，因为它可以用特异性和辐射质量的生物标记物来丰富。这 应用程序还结合了我们的协作者的工程经验和能力，以进一步开发 前处理设备，目的是在将样品运送到临床设施期间稳定样品。这个 将制造的材料还将致力于丰富与辐射相关的代谢物的生物特征，并 从少量的生物液体(尿液、血清、全血、唾液)中有效地提取它们，作为 稳定的干膜。将这种材料组装在96孔板中将进一步减少样品 缩短了样品制备时间，最大限度地减少了样品制备过程中的人为错误。我们独特的方法来处理 将LC-MS/MS应用与前处理材料相结合，旨在将这项技术从 技术开发进入可行性阶段，满足辐射损伤快速检测方法的需求 评估。 好了！

This proposal builds upon our metabolomics expertise in untargeted and targeted metabolomics for the generation of highly sensitive and quantitative targeted multiplex assays. The idea behind generating such assays for radiation assessment and radiation injury in easily accessible biofluids (urine, blood, saliva) is to rapidly determine the extent of exposure of an individual and distinguish between the worried well and the exposed individuals that may require medical intervention. Highly quantitative approaches will be undertaken through liquid chromatography tandem mass spectrometry (LC-MS/MS) to quantify each already identified radiation biomarkers in each biofluid. Such instruments are currently used routinely in clinical laboratories, therefore maximizing the available resources for rapid evaluation of thousands of individuals during an emergency. Based on criteria for sensitivity, high signal-to-noise ratio, low signal suppression from matrix effects, and high fold changes compared to controls or relationships between pairs of metabolites, biosignatures will be assembled and concentrations calculated. The combined biosignature will be developed in a multiplex assay, effectively reducing the time between sample preparation to results. The goal is to demonstrate that this multiplex assay method has the potential to be deployed in the case of an emergency to a pre-determined network of clinical laboratories that can accept and rapidly process a high volume of samples. While the ultimate goal will be for such a panel to be predictive in all cases, even a limited false positive rate would facilitate assessment of radiation injury in a mass casualty scenario: e.g. a 1% false positive rate would reduce the number of individuals needing further evaluation by 100-fold. Additionally, this assay will be flexible as it could be enriched with biomarkers for specificity and radiation quality. This application also combines the engineering experience and capabilities of our collaborators to further develop pre-processing devices with the intention of stabilizing the sample during transport to a clinical facility. The materials to be fabricated will also aim to enrich the biosignature for the radiation-related metabolites and extract them effectively from small amounts of a biofluid (urine, serum, whole blood, saliva), transported as a stable dry membrane. Assembly of such materials in a 96 well plate will further decrease the sample preparation time and minimize human error associated with sample preparation. Our unique approach to combine LC-MS/MS applications with pre-processing materials will aim to move this technology from the feasibility stage to technology development, satisfying the needs for rapid methods for radiation injury assessment. !