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

描述（由申请人提供）：本提案的目的是评估一种全集成设备的可行性，该设备用于全血氰化物中毒的快速和早期诊断，使用维生素B12前体cobinamide与氰化物结合时的光谱位移作为指标。氰化物是一种非常有效和迅速起作用的毒药，对人类致命的剂量只有50毫克。目前还没有便携式快速检测全血氰化物的方法。该提案的主要目标是通过使用手指棒的全血样本进行快速测试，以证明组合酰胺-氰化物化学的可行性。从样品收集到有效结果的总分析时间将少于5分钟。该提案的创新之处在于将大麻酰胺化学与血液分离技术、流体路径设计和检测区域结合在一起，所有这些都集成在一个紧凑的一次性设备中，可以用手持可视化系统进行解释。这种新型快速检测血液氰化物的可行性工作将首先集中在设备设计的几种技术方法上。追求湿化学设计的平行路径，试剂在设备中干燥的设计，以及它们的组合将被考虑，比较和评估。在所有提出的方法中，全血样本将被添加到设备中，随后是一个集成的全血处理步骤，以产生血浆样本。在全血分离和裂解之前或之后，样品将与机载试剂混合，干燥或潮湿，可能还有缓冲液。然后，处理后的样品-试剂混合物通过各种流体路径通过该装置移动到检测区域。为了使样品和试剂之间有足够的混合时间，将评估不同的流体路径、混合室、聚合物时间门和可溶解膜，以最大限度地提高相互作用时间。将在检测区域使用手持或小型便携式读取器测量氰化物与cobinamide结合时cobinamide的光谱位移。在成功完成第一阶段项目后，我们打算进一步开发我们的概念/原型分析，完成设备设计，并通过第二阶段提案将完成的测试与阅读器仪器集成。该项目第二阶段的具体目标包括验证以组合胺为基础的测量血液中氰化物的装置。结果将与测定生物样品中氰化物的两种既定方法进行比较。将确定设备的灵敏度、特异性、准确性、精密度和可重复性。我们将设计更多的研究以获得监管机构的批准，随后我们将商业化该测试。如果成功，这项测试将为医学界和急救人员提供一种快速、可靠和经济的方法来确定氰化物中毒。

DESCRIPTION (provided by applicant): The objective of this proposal is to assess the feasibility of a fully integrated device for the rapid and early diagnosis of cyanide poisoning in whole blood using the spectral shift of the vitamin B12 precursor cobinamide upon binding with cyanide as an indicator. Cyanide is an extremely potent and rapid acting poison with as little as 50 mg fatal to humans. Currently there are no portable rapid tests for the detection of cyanide in whole blood available. The primary goal of this proposal is to demonstrate feasibility of the cobinamide-cyanide chemistry in a rapid test using a whole blood sample from a finger-stick. The total assay time from the sample collection to a valid result will be less than 5 minutes. The innovation in this proposal is to incorporate the cobinamide chemistry with blood separation technology, fluid path designs, and a detection area, all integrated in a compact, disposable device which can be interpreted with a handheld visualization system. Feasibility work of this novel rapid assay for detecting blood cyanide through cobinamide will initially focus on several technical approaches for the device design. Parallel paths to pursue a wet chemistry design, a design where the reagents are dried in the device, and a combination thereof will be considered, compared and evaluated. In all of the proposed methods the whole blood sample will be added to the device, followed by an integrated whole blood processing step to yield a plasma sample. Either prior to or immediately after the whole blood separation and lysis, the sample will be mixed with the on-board reagent, dried or wet, and possibly buffers. The processed sample-reagent mixture then moves through the device through various fluid paths to a detection area. To allow for sufficient mixing time between the sample and the reagents, different fluid paths, mixing chambers, polymeric time-gates, and dissolvable films will be evaluated with the goal of maximizing the interaction time. The spectral shift of cobinamide upon binding of cyanide to cobinamide will be measured in the detection area using a handheld or small, portable reader instrument. Upon successful completion of this Phase 1 project we intend to further develop our concept/prototype assay, complete the device design and integrate the finished test with a reader instrument through a Phase 2 proposal. Specific aims for a Phase 2 of this project include the validation of the cobinamide-based device for measuring cyanide in blood. Results will be compared to two established methods for measuring cyanide in biological samples. The sensitivity, specificity, accuracy, precision, and reproducibility of the devices will be determined. Additional studies will be designed to obtain regulatory approval and subsequently we will commercialize the test. If successful, this test will provide the medical community and first responders with a fast, reliable and economic alternative to determine cyanide poisoning. PUBLIC HEALTH RELEVANCE: Cyanide poisoning has been recognized as a threat from smoke inhalation and potentially through weapons of mass destruction, but there are currently no methods available to rapidly detect cyanide in blood from at-risk patients. The objective of the proposed work is to assess the feasibility of a fully integrated method for the rapid and early detection and diagnosis of cyanide poisoning in whole blood using the spectral shift of the vitamin B12 precursor cobinamide upon binding with cyanide as an indicator. Successful accomplishment of the goals of this project will be the basis for the development of a test to detect and measure cyanide blood poisoning which will allow healthcare providers to treat patients quickly and effectively.