A Rapid and Sensitive Test for Gamma-hydroxybutyrate

描述（由申请方提供）：将通过使用γ-羟基丁酸脱氢酶（GHB-DH）的特定酶法开发一种简单易用的尿液检测试纸。拟议的第一阶段研究将确定将不稳定的酶液体试剂形式转化为环境温度（20 - 25摄氏度）稳定的固相形式的可行性。具体目标是：（1）选择试剂处理材料和方法;（2）确定用于保护测试条中的生化物质的稳定剂和工艺;以及（3）构建和评估测试条的性能。为了证明可行性，干燥试剂垫必须具有在660 nm处反射率大于95%的近白色背景色，初始环境温度稳定性为一个月或更长时间，而液体试剂稳定性小于3小时，并且当以试纸形式放置时，在不到2分钟内运行测试并检测至少0.1 mg/mL GHB的样品。我们希望识别出具有低背景颜色（高反射率）且不含与所用酶反应的添加剂的试剂垫材料（目标1）。将尝试使用各种稳定剂，以确定在干燥过程中和干燥后保护酶的稳定剂。此外，将尝试不同的干燥条件，以确定产生最稳定的干燥垫的干燥条件（目标2）。最后，我们将测试和评估原型带，以确定它们是否符合可行性标准（目标3）。开展该项目是因为迫切需要一种能够快速检测体液中γ-羟基丁酸的诊断试剂盒。急诊室医生需要一种可靠和快速的检测方法来评估昏迷状态的患者。目前，他们没有这样的工具。除了ER应用外，该测试还具有潜在的执法应用。估计全球市场每年超过6000万美元。

DESCRIPTION (provided by applicant): A simple-to-use urine test strip is to be developed from a specific enzymatic method that uses gammahydroxybutyrate dehydrogenase (GHB-DH). The proposed Phase I research will determine the feasibility of converting the unstable enzymatic liquid reagent format to an ambient temperature (20 - 25 ¿ Celsius) stable solid phase format. The specific aims are: (1) select reagent process materials and method; (2) identify stabilizers and processes for protecting the biochemicals in the test strip; and (3) build and evaluate the performance of test strips. To demonstrate feasibility, the dried reagent pad must have a near-white background color of greater than 95% reflectance at 660 nm, an initial ambient temperature stability of one month or more versus the liquid reagent stability of less than 3 hours, and when placed in a strip format, run a test in less than 2 minutes and detect a sample of at least 0.1 mg/mL GHB. We expect to identify materials for the reagent pad that have low background color (high reflectance) and have no additives that will react with the applied enzymes (aim 1). A variety of stabilizing agents will be tried to identify those that will protect the enzymes during and after the drying process. Additionally, different drying conditions will be tried to identify the one that yields the most stable dried pad (aim 2). Finally, we will test and evaluate prototype strips to determine if they meet the feasibility criteria (aim 3). The project is being undertaken due to an urgent need for a diagnostic kit that can rapidly detect gammahydroxybutyrate in bodily fluids. Emergency room physicians need a reliable and rapid assay for evaluating patients that arrive in a comatose state. Currently, they do not have such a tool at their disposal. In addition to an ER application, the test also has potential law enforcement applications. The estimated world wide market is over $60 million per year.