Curbside Immunoassay Device for Quantitative Measurement of Cannabis Intoxication

用于定量测量大麻中毒的路边免疫测定装置

• 批准号: 9141902
• 负责人: MICHAEL JOHN HEFFERNAN
• 金额: $ 22.44万
• 依托单位: BREVITEST TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9141902
• 关键词: Air; Antibodies; Area; Biocompatible Materials; Biological Assay; Biotin; Blood; Blood specimen; Buffers; Cannabis; Caymans; Chemicals; Collaborations; Detection; Devices; Diagnosis; Drug Addiction; Drug Kinetics; Drug abuse; Enzymes; Exhibits; Foundations; Half-Life; Hour; Immunoassay; Intoxication; Laboratories; Lateral; Law Enforcement; Link; Liquid substance; Logistics; Magnetic nanoparticles; Marijuana; Measurement; Measures; Methods; Microfluidics; Movement; Optics; Oral; Pattern; Phase; Reaction; Reagent; Reproducibility; Research Personnel; Robotics; Route; Running; Safety; Saliva; Sampling; Sensitivity and Specificity; Serum; Site; Small Business Innovation Research Grant; Statutes and Laws; Streptavidin; Substance Use Disorder; THC concentration; Technology; Testing; Tetrahydrocannabinol; Time; Urine; Validation; Work; Workplace; antibody conjugate; base; design; drug testing; field study; innovation; innovative technologies; marijuana legalization; marijuana use; meter; minimally invasive; novel; particle; phase 2 study; programs; public health relevance; research clinical testing; tool

 DESCRIPTION (provided by applicant): Law enforcement officials, workplace drug testing programs, and researchers in the field of drug abuse lack rapid, easily deployable tools to measure present cannabis (marijuana) intoxication. The enzyme-linked immunosorbant assay (ELISA) is an established cannabis testing method for detecting delta-9-tetrahydrocannabinol (THC) levels in urine or blood; however, the logistics of obtaining urine or blood samples on site represents a major limitation of current THC ELISAs, particularly for curbside testing of drivers. The most feasible candidate sample for curbside testing is the oral fluid, as it is a minimally invasive sampling route and the pharmacokinetics of THC concentration in oral fluid (half-life of 1.5 hours) parallels that of serum; however, the primary obstacle to this approach is the lack of a rapid, quantitative THC immunoassay. Traditional plate-based ELISA tests can take 1 to 3 hours and require a laboratory setting, and lateral flow assays lack the accuracy needed for quantitative cannabis intoxication detection. BreviTest Technologies LLC is developing a portable analyzer to perform a rapid, quantitative ELISA test within a 10-minute run time. The sample is loaded into a discontinuous microfluidics cartridge, and a robotic mechanism controls the movement of antibody-conjugated magnetic nanoparticles through successive fluid wells, where enhanced mixing patterns substantially reduce the time required for each ELISA step. This innovative approach enables the implementation of a rapid, portable, fully automated ELISA test with accuracy comparable to a standard laboratory ELISA. In this SBIR Phase I project, BreviTest will partner with Cayman Chemical Company to develop a THC ELISA test using the BreviTest robotic analyzer, with the following Specific Aims: 1) Develop and produce discontinuous microfluidic cartridges with layman-operable sample port. The type and quantity of magnetic nanoparticles will be determined for optimal reagent mixing and transit parameters, and the design and validation will be completed for a sampling tool for metering 20 µL of sample fluid. Design of the cartridge will be optimized for stability, consistency, and compatibility with biological materials. 2) Develop curbside ELISA test to quantitatively measure THC in oral fluid. BreviTest will collaborate with Cayman Chemical to screen commercially available clones to generate an optimized antibody for THC detection. Antibodies will be conjugated to magnetic nanoparticles via streptavidin-biotin linkages, for use in the BreviTest analyzer. Optimization of reagent concentrations and analyzer parameters will be performed, along with statistical validation of dynamic range, specificity, and sensitivity. The completion of these proof-of-concept studies will demonstrate the ability of the BreviTest portable ELISA analyzer to measure THC concentration in oral fluid samples, at concentrations relevant to curbside intoxication analysis. This work will lay the foundation for further Phase II studies including fied and clinical testing of a rapid THC intoxication assay and additional validation testing towards a defined 510(k) clearance by the FDA.

 描述（由申请人提供）：执法官员、工作场所药物检测项目和药物滥用领域的研究人员缺乏快速、易于部署的工具来测量当前的大麻（大麻）中毒情况。建立了检测尿液或血液中 delta-9-四氢大麻酚 (THC) 水平的大麻测试方法；然而，现场获取尿液或血液样本的后勤工作是当前 THC 的主要限制。 ELISA，特别是对于驾驶员的路边测试，路边测试最可行的候选样本是口腔液，因为它是一种微创采样途径，并且口腔液中 THC 浓度的药代动力学（半衰期为 1.5 小时）与口腔液中的 THC 浓度相似。然而，这种方法的主要障碍是缺乏快速、定量的 THC 免疫测定，传统的基于板的 ELISA 测试可能需要 1 至 3 小时，并且需要实验室环境，而侧流测定则缺乏这种方法。 BreviTest Technologies LLC 正在开发一种便携式分析仪，可在 10 分钟运行时间内执行快速定量 ELISA 测试，将样品装入不连续的微流体盒中，并由机器人机构控制其运动。抗体偶联的磁性纳米粒子通过连续的流体孔，其中增强的混合模式大大减少了每个 ELISA 步骤所需的时间，这种创新方法能够准确地实施快速、便携式、全自动 ELISA 测试。在这个 SBIR 第一阶段项目中，BreviTest 将与开曼化学公司合作，使用 BreviTest 机器人分析仪开发 THC ELISA 测试，具体目标如下： 1) 开发和生产非连续微流体盒将确定磁性纳米粒子的类型和数量以获得最佳试剂混合和运输参数，并完成采样工具的设计和验证。计量 20 µL 样品液的设计将针对稳定性、一致性和兼容性进行优化。 2) 开发路边 ELISA 测试，以抗体测量口腔液中的 THC，BreviTest 将与 Cayman Chemical 合作筛选市售克隆，以生成针对 THC 检测的优化抗体将通过链霉亲和素-生物素连接与磁性纳米粒子结合。将优化试剂浓度和分析仪参数，并对动态范围、特异性和灵敏度进行统计验证。这些概念验证研究的完成将证明 BreviTest 便携式 ELISA 分析仪能够测量口腔液样本中 THC 浓度，其浓度与路边中毒分析相关。这项工作将为进一步的 II 期研究（包括现场研究）奠定基础。快速 THC 中毒测定的临床测试以及针对 FDA 定义的 510(k) 许可的额外验证测试。