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

描述)：这项提议的目标是完成一种现场可部署的、完全集成的设备的开发，用于快速和早期诊断全血中的氰化物中毒。维生素B12前体可比胺与氰化物结合后的特定光谱位移将用于测量样品中氰化物的量。氰化物是一种威力极强、见效迅速的毒药，对人类的致命性只有50毫克。目前还没有便携的快速检测全血中氰化物的方法。然而，急救人员和急救人员需要在现场快速测量全血中的氰化物水平，以便对接触过氰化物的人进行解毒剂治疗，我们的测试旨在填补这一空白。在可行性过程中，我们证明了使用我们的概念设备可以检测到兔全血中低水平的氰化物。我们通过在膜上固定改性的可比酰胺--可比酰胺聚合物，将可比酰胺化学集成到这个原型装置中。为了实现这一点，我们必须开发一种化学修饰，通过这种化学修饰，当氰化物与聚合物连接后，钴酰胺在结合氰化物时保持其特有的光谱位移。在我们的原型装置中，全血被裂解，然后通过固定的可比酰胺聚合物，然后进行洗涤步骤。用手持式分光光度计测量了可比酰胺与氰化物结合后的光谱位移。我们现在建议完成这一原型设备的开发，使其成为功能齐全的集成测试设备。首先，我们将最终确定所有的检测组件，并优化用于测试的化学成分 以最大限度地提高分析灵敏度。与此同时，我们将开发一个样本集合 该模块将使我们能够重复采集测量数量的全血，裂解红细胞，并将样本送到测试设备。一旦仪器和样品模块达到原型阶段，我们就开始开发一种便携式仪器，它可以解释测试结果，并向用户提供样品中氰化物的浓度。原型设备将进行校准，然后使用接触氰化物的兔子和猪的样本进行评估，这些动物模型已经证明成功，并已被科学和监管机构接受。最终将对生产设备、样品采集模块和仪器进行广泛的测试和验证。这一阶段的验证研究将包括确认敏感性、特异性、准确性、精密度和重复性，以及FDA批准通常所需的其他研究。临床研究，包括关键的动物研究，不在本项目的范围内，将在本阶段完成后启动。

DESCRIPTION): The objective of this proposal is to complete the development of a field-deployable, fully integrated device for the rapid and early diagnosis of cyanide poisoning in whole blood. The specific spectral shift of the vitamin B12 precursor cobinamide after binding cyanide will be used to measure the amount of cyanide in the sample. 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. However, emergency personnel and first responders have a need to measure the levels of cyanide in whole blood quickly on site in order to treat exposed individuals with an antidote and our test is intended to fill that niche. During feasibility, we demonstrated that we can detect low levels of cyanide in whole blood from rabbits with our concept device. We integrated the cobinamide chemistry into this prototype device by immobilizing modified cobinamide, cobinamide-polymer, on a membrane. To achieve that we had to develop a chemical modification through which the cobinamide retains its characteristic spectral shift when binding cyanide after it has been linked to a polymer. In our prototype device, whole blood was lysed, and then passed through the immobilized cobinamide-polymer, followed by a wash step. The spectral shift of cobinamide upon binding cyanide was measured with a handheld spectrophotometer. We now propose to complete the development of this prototype device into a fully functional and integrated test device. First, we will finalize all the assay components and optimize the chemistry for the test in order to maximize the analytical sensitivity. In parallel we will be developing a sample collection module that will enable us to reproducibly collect a measured amount of whole blood, lyse the red cells and deliver the sample to the test device. Once the device and the sample module have reached the prototype stage, we are initiating the development of a portable instrument which can interpret the test and provide the user with a concentration of cyanide in the sample. The prototype devices will be calibrated and then evaluated with samples from cyanide exposed rabbits and pigs, animal models that have documented success and have been accepted by the scientific and regulatory community. Production devices, sample collection modules and instruments will finally be extensively tested and validated. Validation studies for this Phase wil include the confirmation of sensitivity, specificity, accuracy, precision, and reproducibility and other studies that are typically required for FDA clearance. Clinical studies, including pivotal animal studies are outside the scope of this project and will be initiated upon completion of this Phase.