Development of a Field-Deployable Device to Rapidly Measure Blood Cyanide Levels

项目摘要 这项提议的目的是完成一个可在外地部署、 用于快速和早期诊断全血中氰化物中毒的全集成传感器。 氰化物是一种非常有效和快速的毒药。它可以被吸入或通过皮肤吸收为氰化物 气体或作为氰化物盐摄入，并具有多种毒性机制。吸入含氰化物的烟雾 是住宅火灾中氰化物致人伤亡的主要原因之一。氰化物中毒的解毒剂有 目前，虽然价格很高。虽然新的和更有效的治疗剂在 发展，没有快速诊断测试，以支持应急人员和第一反应谁有一个 需要快速测量全血中的氰化物水平，以便安全有效地治疗暴露个体。 我们的测试旨在填补这一空白。 我们建议完成我们的原型传感器的发展成为一个功能齐全的综合测试设备 并验证其性能。我们已经建立并证明了化学品的化学性质和功能 传感器，我们已经将传感器集成到一个微流体盒中，该微流体盒收集测量的血液量 它包含所有的试剂和缓冲液，并容纳氰化物传感器。在我们的原型中 收集固定量的全血样品，溶解红细胞，然后 通过传感器，然后是手动驱动的洗涤步骤。在此阶段，我们将首先完成 将氰化物传感器和裂解/洗涤缓冲液试剂整合到微流体盒中 改进和优化墨盒，以解决剩余的性能挑战。测试合格 并验证一次性耗材最初将使用我们的原型台式读取器完成。与此同时，我们的主要努力 将专注于手持阅读器的开发。在概念阶段之后， 将开发仪器并使用试剂盒进行测试，以确认功能并确定 改进.此外，阿尔法阅读器将在一项可用性研究中与预期用户、消防员 和EMT的，以评估他们的表现，并获得反馈，以改善设计和功能的 系统我们的阅读器的硬件和软件开发然后按计划进展到预生产 在项目结束之前，进行广泛的完整系统测试。预生产系统 性能，一次性用品和仪器，将彻底评估与氰化物暴露的样品 兔子和猪，作为动物模型，以及掺入人血。本阶段的验证研究将包括 确认灵敏度/特异性（≥95%）、检测限（≤20 µM）、动态范围（20-150 µM）， 准确度/重现性（≤20%变异性）、测定时间（≤5分钟）和通常需要的其他研究 等待FDA批准本阶段完成后，将启动生产等效试验的临床研究。

PROJECT SUMMARY The objective of this proposal is to complete the development and verify the performance of a field-deployable, fully integrated sensor for the rapid and early diagnosis of cyanide poisoning in whole blood. Cyanide is an extremely potent and rapid acting poison. It can be inhaled or absorbed across the skin as cyanide gas or ingested as cyanide salt, and has multiple mechanisms of toxicity. Smoke inhalation containing cyanide is one of the main causes of cyanide related casualties in residential fires. Antidotes to cyanide poisoning are currently available, although at a high price. While new and more effective therapeutic agents are in development, there is no rapid diagnostic test to support emergency personnel and first responders who have a need to measure cyanide levels in whole blood quickly in order to safely and effectively treat exposed individuals. Our test is intended to fill that niche. We propose to complete the development of our prototype sensor into a fully functional and integrated test device and to verify its performance. We have established and proven the chemistry and functionality of the chemical sensor and we have integrated the sensor into a microfluidic cartridge that collects a measured amount of blood from a finger-stick, contains all the reagents and buffers, and houses the cyanide sensor. In our prototype microfluidic cartridge a fixed amount of a whole blood sample is collected, the red blood cells are lysed, then passed through the sensor, followed by a manually actuated wash step. In this phase we will first complete the integration of the cyanide sensor and lysis/wash buffer reagents into the microfluidic cartridge after making improvements to and optimizing the cartridge to resolve remaining performance challenges. Testing to qualify and verify the disposable will be done initially with our prototype benchtop reader. In parallel, our main efforts will focus on the hand-held reader development. After a concept stage, alpha versions of the field-deployable instrument will be developed and tested with the cartridge to confirm functionality and to identify areas for improvement. In addition, alpha readers will be evaluated in a usability study with intended users, firefighters and EMT’s, to evaluate their performance and obtain feedback to improve the design and functionality of the system. Hardware and software development of our reader are then scheduled to progress to a pre-production stage and undergo extensive complete system testing prior to the end of this project. The preproduction system performance, disposable and instrument, will be thoroughly evaluated with samples from cyanide exposed rabbits and pigs, as animal models, and spiked human blood. Verification studies for this Phase will include the confirmation of sensitivity/specificity (≥95%), limit of detection (≤20 µM), dynamic range (20-150 µM), accuracy/reproducibility (≤20% variability), assay time (≤5 minutes), and other studies that are typically required for FDA approval. Clinical studies with production equivalent tests will be initiated upon completion of this Phase.