A Rapid and Sensitive Test for Gamma-hydroxybutyrate

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 x 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.