Rapid tests for the detection of inhibitors of enzymes and human exposure to the same

Inventors

Vallejo, JR., Yli Remo • Milunic, David

Assignees

Fulgent Genetics Inc

Abstract

A device and method for the rapid on-site detection of inhibitors of enzymes, such as, acetylcholinesterase is described where the device contains 2 reaction zones containing a reporter enzyme substrate. One reaction zone is for the test sample while the other is for an onboard negative control. Sample and control fluids are preincubated with the enzyme in separate reaction containers, then an aliquot of each reaction mixture is added to designated reaction zones on the test device. A purpose-built reader or an illuminating device, such as, containing an incandescent light source, a diode, a UV light source or any other illumination source that is suitable for the reporter or mere visualization is used to determine the level of reporter.

Core Innovation

The invention relates to a system for detecting presence of an inhibitor of an esterase. The system includes a self-calibrated, modular, handheld device with a top piece and a bottom piece, where the bottom piece comprises a negative control reaction zone and a test reaction zone positioned below openings in the top piece when assembled. Each reaction zone comprises a porous matrix consisting essentially of an immobilized substrate of the esterase, and the negative control reaction zone and the test reaction zone are separate and exclusive.

In the reaction of the immobilized substrate and the esterase, a product of the reaction comprises a detectable reporter. A negative control reaction zone includes a negative control sample comprising detectable reporter, enabling comparison between the negative control reaction zone and the test reaction zone for a test sample suspected of comprising an inhibitor of an esterase. The invention further includes, separate from the device, two reaction containers, each container comprising a same amount of the esterase.

Additionally, the system includes a reader that detects the detectable reporter. The detected reporter from the test reaction zone is assessed relative to the negative control reaction zone, using the reader’s detection outputs to generate a fluorescence, colorimetric, or luminogenic readout and to support positive/negative determination, including use of ratio/threshold algorithms and avoidance of sample oxidation by using carboxylesterase in place of or in addition to acetylcholinesterase.

Claims Coverage

The disclosed material contains one independent claim. Its inventive coverage centers on a self-calibrated modular handheld device with separate, exclusive negative control and test reaction zones using porous matrices with immobilized esterase substrate, together with separate reaction containers holding matched esterase amounts and a reader to detect a reporter produced by esterase activity, for determining presence of an esterase inhibitor.

Overall claim coverage is directed to a self-calibrated modular handheld assay platform that compares reporter outputs between a negative control reaction zone and a separate test reaction zone using porous matrices with immobilized esterase substrate, where matched esterase is provided in two reaction containers and a reader detects the detectable reporter to determine presence of an esterase inhibitor.

Stated Advantages

Documented Applications