Bench-top Reader and Aptamer-based Assay for Rapid, High-sensitivity Drug/Opiate Detection

项目总结/摘要 美国正在经历一场规模空前的流行病，海洛因的使用死灰复燃。 以及滥用处方阿片类药物如芬太尼，芬太尼， 衍生物、美沙酮、羟考酮和氢可酮。每10个药物过量中就有6个是 现在与阿片类药物有关，导致每天约90人死亡，每年的经济影响超过750亿美元。 此外，对检测许多其他小分子的需求迅速扩大， 治疗剂，在家里或潜在的大规模杀伤性武器，如沙林和有毒农药在偏远地区 或字段设置。为了开发一种通用工具来解决这种检测问题，我们建议改进 我们的补偿干涉读取器（CIR），并进一步开发了一个独特的和有效的，矩阵 不敏感的测定方法，以满足对阿片类药物的高灵敏度、快速检测的需求， 治疗剂和环境污染物。我们的努力将产生第一个这样的长凳- 顶级（和现场兼容）CIR和用户友好（交钥匙）检测系统，用于检测和/或 以ng/mL-pg/mL灵敏度定量广泛的靶点，包括阿片类药物、小剂量 分子药物和有机磷农药。基于近表面分析的分析 (NSA)在一个实施例中，由三个步骤组成：1）收集样品（如果需要，则稀释），2）填充预填充物， 涂覆的毛细管和3）用CIR读取“样品”。药物或代谢物检测结果 通过CIR测定的尿对芬太尼家族的敏感性预测为<300 pg/mL，a >10-1000倍 与最佳侧流试验（LFT）1和标准药物试验的当前截止值相比有所改进。 非侵入性、纳升体积、高特异性尿液或缓冲液-稀释剂测定将有助于 对环境样本、缉获的非法麻醉品、假药进行快速实地检测 有机磷神经毒剂（OPNA）。类似的微创测定是可行的， 微量的手指血这种新颖的方法论是通过三个方面的结合实现的。 专利技术，用于适体选择的多重SELEX方法， 测定和CIR以定量靶。第一阶段和第二阶段的目标旨在 使所提出的技术能够快速转化/商业化。

Project Summary/Abstract The United States is experiencing an epidemic of unprecedented scope with heroin use resurging and a troubling pervasive increase in abuse of prescription opioids such as fentanyl, fentanyl derivatives, methadone, oxycodone and hydrocodone. More than 6 out of 10 drug overdoses are now opioid-related, resulting in ~90 deaths/day and a more than $75B/year economic impact. Additionally, there is a rapidly expanding need to detect numerous other small molecules such as therapeutic agents, in the home or potential WMDs like sarin and toxic pesticides in the remote or field setting. To develop a general purpose tool to address such detection, we propose to refine our compensated interferometric reader (CIR) and further develop a unique and efficient, matrix insensitive, assay methodology to address the need for high sensitivity, rapid testing of opioids, therapeutics and environmental contaminants. Our efforts will produce a first of its kind bench- top (and field-compatible) CIR and user-friendly (turn-key) assay system for the detection and/or quantification at ng/mL-pg/mL sensitivity of a wide range of targets including opiates, small molecule drugs, and organophosphorus pesticides. The assay based on a near-surface assay (NSA) embodiment, consists of three-steps; 1) collect sample (dilute if needed), 2) fill the pre- coated capillary and 3) read the ‘sample’ with CIR. Results for drug or metabolite detection in urine by CIR is predicted to be <300 pg/mL sensitivity for the fentanyl family, a >10-1000-fold improvement over the best lateral flow tests (LFT)1 and the current cutoff for standard drug tests. The non-invasive, nanoliter volume, high specificity urine or buffer-diluent assays will facilitate rapid field-based detection of environmental samples, seized illicit narcotics, counterfeit drugs and organophosphorus nerve agents (OPNA). Similar minimally invasive assays are feasible with minute amounts of finger-stick blood. The novel methodology is enabled by the marriage of three patented technologies, a multiplex-SELEX approach for aptamer selection, an aptamer-probe assay, and CIR to quantify the target. Phase I and Phase II Aims proposed here are designed to enable rapid translation/commercialization of the proposed technology.