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

描述（由申请方提供）：本提案的目的是评估一种完全集成的设备的可行性，该设备用于快速和早期诊断全血中的氰化物中毒，使用维生素B12前体可宾酰胺在与氰化物结合时的光谱位移作为指示剂。氰化物是一种非常有效和快速作用的毒药，对人类来说只有50毫克致命。目前还没有用于检测全血中氰化物的便携式快速测试。本提案的主要目的是证明在使用手指针刺全血样本进行快速检测时，Cobinamide-氰化物化学的可行性。从样本采集到有效结果的总测定时间将少于5分钟。该提案的创新之处在于将可宾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，该设备可以用手持式可视化系统进行解释。这种新的快速检测血氰化物的可行性工作，通过cobinamide最初将集中在几个技术方法的设备设计。将考虑、比较和评估追求湿化学设计的平行路径、试剂在设备中干燥的设计及其组合。在所有提出的方法中，将全血样本添加到器械中，然后进行整合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立即将样本与机载试剂（干燥或湿润）以及可能的缓冲液混合。然后，处理过的样品-试剂混合物通过各种流体路径移动通过装置到达检测区域。为了使样品和试剂之间有足够的混合时间，将对不同的流体路径、混合室、聚合物时间门和可溶解膜进行评价，以最大限度地延长相互作用时间。使用手持式或小型便携式读数仪在检测区域测量氰化物与氯苯氨酰胺结合后氯苯氨酰胺的光谱位移。在成功完成第1阶段项目后，我们打算进一步开发我们的概念/原型试验，完成器械设计，并通过第2阶段提案将完成的测试与读数仪仪器集成。该项目第2阶段的具体目标包括验证用于测量血液中氰化物的基于钴胺的设备。结果将比较两个既定的方法来测量生物样品中的氰化物。将确定器械的灵敏度、特异性、准确度、精密度和再现性。将设计更多的研究以获得监管机构的批准，随后我们将使该测试商业化。如果成功，该测试将为医学界和急救人员提供一种快速，可靠和经济的替代方法来确定氰化物中毒。 公共卫生相关性：氰化物中毒被认为是吸入烟雾和可能通过大规模杀伤性武器造成的威胁，但目前还没有方法可以快速检测高危患者血液中的氰化物。拟议工作的目的是评估一个完全集成的方法的可行性，用于快速和早期检测和诊断全血中的氰化物中毒，使用维生素B12前体cobinamide与氰化物结合后的光谱位移作为指标。该项目目标的成功实现将是开发检测和测量氰化物血液中毒的测试的基础，这将使医疗保健提供者能够快速有效地治疗患者。

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.