Sample-free onsite detection of cocaine using microneedles via laser-treated skin

使用微针通过激光处理的皮肤对可卡因进行无样品现场检测

• 批准号: 9044652
• 负责人: Mei X Wu
• 金额: $ 22.36万
• 依托单位: AFASCI, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-15 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9044652
• 关键词: Accident and Emergency department; Administrator; Animals; Area; Binding; Biological; Biological Assay; Biological Markers; Blood; Blood specimen; Boxing; Capillary Permeability; Cessation of life; Clinic; Clinical; Cocaine; Cocaine-Related Disorders; Collection; Color; Communities; Country; Crime; DNA; Data Analyses; Dermal; Dermis; Detection; Devices; Diagnosis; Dimensions; Dose; Drug Controls; Drug abuse; Economics; Emergency Situation; Emergency department visit; Engineering; Equipment; Ethical Issues; Family suidae; Feedback; Fluorescein-5-isothiocyanate; Fluorescence; General Hospitals; Goals; Gold; Grant; Hemoglobin; Heroin; Hospitals; Human; Illicit Drugs; In Situ; In Vitro; Institutes; Label; Lasers; Lead; Length; Light; Lighting; Marijuana; Massachusetts; Measurement; Measures; Medical; Methods; Modification; Mus; Needles; Oranges; Painless; Penetration; Pharmaceutical Preparations; Phase; Plasma; Policies; Protocols documentation; Public Health; Rattus; Reaction Time; Research; Risk; Rodent; Saliva; Sampling; Sensitivity and Specificity; Series; Shapes; Skin; Small Business Innovation Research Grant; Small Business Technology Transfer Research; Specificity; Specimen; Surface; Technology; Testing; Transdermal substance administration; United States; United States National Institutes of Health; Urinalysis; Urine; Variant; Visit; Workplace; analytical method; animal facility; aptamer; base; benzoylecgonine; cocaine use; commercialization; cost; cyanine dye 5; design; drug testing; drugged driving; experience; improved; in vitro Assay; in vitro testing; in vivo; injured; minimally invasive; novel; prevent; product development; protocol development; public health relevance; receptor; sample collection; screening; sensor; signal processing; social; specific biomarkers; success

 DESCRIPTION (provided by applicant): The abuse of illicit drugs such as cocaine, marijuana and heroin remains a critical public health concern throughout the country and is associated with staggering economic and social consequences. Cocaine remains the most frequently recorded illicit drug in emergency hospital visits and the leading cause of drug- related deaths in the United States. Since as high as 23% of fatally injured drivers were found positive results in drug tests, the Office of National Drug Control Policy emphasizes the major concern of drugged driving in the United States. Urinalysis is the most common type of cocaine test, though urine collection has challenged its practicality. There are other biological specimen-based cocaine tests that have been explored such as blood and saliva tests but are not practical since blood samples need high-level medical administrators and specialized analytic equipment. Another alternative is the minimally invasive diagnosis through skin using surface modified microneedles (MNs). However, since concentrations of most blood biomarkers in the skin are too low to be captured by the MN array, the test suffers from low sensitivity and high variation. Here we propose a novel probe-MN-based assay in conjunction of using brief laser illumination on a tiny skin area to accumulate blood biomarker, such as cocaine into the upper dermis. This sample accumulation method enables the biomarker to be readily detected and quantified in situ using the probe-MNs inserted into the laser- treated skin. The objective for the STTR Phase I is to characterize and validate sensitivity and specificity of the probe-MN array in vitro and in vivo, and compare detection reliability with conventional analysis of cocaine in plasma and in urine samples. The specific aims are as follows: Aim 1: Determine optimal design of cocaine-specific binding MN probe and characterize cocaine assay in vitro. We will develop a protocol of surface modification of MNs with Cy3-labeled cocaine-specific aptamer and Cy5- labeled neutralizer. Length of the MNs ranging from 50 - 300um will be optimized based on iterative feedback from in vivo test. Sensitivity, specificity, and response time of the MNs array will be evaluated by in vitro tests. Aim 2: Demonstrate the feasibility and consistence of upper dermal cocaine detection in mice and rats. The cocaine-specific MN probe will be evaluated in mice and rats to demonstrate feasibility of the proposed minimally invasive MN-based cocaine detection in conjunction of using a brief illumination of a clinically safe dose of laser on the skin. Sensitiviy, specificity, and reliability will be determined with respect to conventional assays of plasma and urinalysis of cocaine. The ultimate goal of the entire STTR project is to develop and commercialize the minimally invasive, sample- free, in situ cocaine detection toolkit consisting of device and cocaine-specific probe-MN arrays for prompt onsite cocaine detection in humans in community, work places, or emergency rooms and clinics.

 描述（由申请人提供）：可卡因、大麻和海洛因等非法药物的滥用仍然是全国范围内的一个重要的公共卫生问题，并与惊人的经济和社会后果相关。可卡因仍然是急诊医院就诊中最常见的非法药物。以及毒品相关死亡的主要原因 美国国家药物管制政策办公室强调，由于高达 23% 的致命伤害司机在药物测试中发现了阳性结果，因此尿液分析是美国最常见的可卡因测试类型。尽管尿液采集对其实用性提出了挑战，但还探索了其他基于生物样本的可卡因测试，例如血液和唾液测试，但由于血液样本需要高级医疗管理人员和专门的分析设备，因此并不实用。通过皮肤进行微创诊断然而，由于皮肤中大多数血液生物标志物的浓度太低，无法被 MN 阵列捕获，因此该测试的灵敏度较低且变异较大。在此，我们提出了一种基于探针的新型检测方法。结合在微小的皮肤区域上使用短暂的激光照射来积累血液生物标志物，例如将可卡因沉积到上层真皮中，这种样本积累方法使得使用插入激光的探针 MN 可以轻松地在原位检测和量化生物标志物。皮肤。 STTR 第一阶段的目标是在体外和体内表征和验证探针-MN 阵列的灵敏度和特异性，并将检测可靠性与血浆和尿液样本中可卡因的常规分析进行比较。具体目标如下： 1：确定可卡因特异性结合 MN 探针的最佳设计，并在体外表征可卡因测定。我们将开发一种使用 Cy3 标记的可卡因特异性适体和 Cy5 标记的中和剂对 MN 进行表面修饰的方案。 50 - 300um 将根据体内测试的迭代反馈进行优化​​ MNs 阵列的灵敏度、特异性和响应时间将通过体外测试进行评估 目标 2：证明小鼠上皮可卡因检测的可行性和一致性。可卡因特异性 MN 探针将在小鼠和大鼠中进行评估，以证明所提出的基于 MN 的微创可卡因检测结合使用临床安全剂量的短暂照明的可行性。激光在皮肤上的敏感性、特异性和可靠性将根据可卡因的常规血浆和尿液分析来确定。整个 STTR 项目的最终目标是开发和商业化微创、无样品、原位分析。可卡因检测工具包包括 设备和可卡因特异性探针 MN 阵列，用于在社区、工作场所或急诊室和诊所对人体进行快速现场可卡因检测。