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

项目总结 这项提议的目标是完成研制并验证可现场部署的、 全集成传感器，用于全血氰化物中毒的快速早期诊断。 氰化物是一种效力极强、见效迅速的毒药。它可以被吸入或以氰化物的形式通过皮肤吸收 气体或以氰化盐的形式摄入，具有多种毒性机制。含氰化物烟雾吸入 是住宅火灾中与氰化物有关的伤亡的主要原因之一。氰化物中毒的解毒剂是 目前有售，尽管价格很高。当新的更有效的治疗药物问世时 在开发过程中，没有快速诊断测试来支持急救人员和具有 需要快速测量全血中的氰化物水平，以便安全有效地治疗受影响的个人。 我们的测试旨在填补这一空白。 我们计划完成我们的传感器原型的开发，使其成为功能齐全的集成测试设备 并验证其性能。我们已经建立并证明了这种化学物质的化学和功能 传感器，我们已经将传感器集成到一个微流体盒中，它可以收集测量到的血量 从一个手指棒，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 微流控芯片采集一定数量的全血样本，裂解红细胞，然后 通过传感器，然后是手动洗涤步骤。在此阶段中，我们将首先完成 在制造后将氰化物传感器和裂解/洗涤缓冲试剂集成到微流控墨盒中 改进和优化盒式磁带，以解决剩余的性能挑战。测试以获得资格 并验证一次性阅读器将首先使用我们的原型台式阅读器完成。同时，我们的主要努力 将重点放在手持阅读器的发展上。在概念阶段之后，阿尔法版本的现场-可部署 将与墨盒一起开发和测试仪器，以确认功能并确定以下区域 进步。此外，Alpha阅读器将在一项针对预期用户、消防员的可用性研究中进行评估 和EMT的，以评估其性能并获得反馈，以改进 系统。我们阅读器的硬件和软件开发将按计划进入试生产阶段 在本项目结束前进行阶段和广泛的完整系统测试。预制系统 性能，一次性和仪器，将通过暴露的氰化物样本进行彻底评估 兔子和猪作为动物模型，并添加了人类血液。这一阶段的核查研究将包括 确认灵敏度/特异度(≥95%)、检出限(≤20微米)、动态范围(20-150微米)、 准确度/重复性(≤20%变异性)、分析时间(≤5分钟)以及通常需要的其他研究 等待FDA批准。在这一阶段结束后，将启动具有生产等值测试的临床研究。

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.