Hydrogen peroxide droplet-based assays
AssigneeAdvanced Liquid Logic, Inc.
InventorsAllen E. Eckhardt, Vijay Srinivasan, Vamsee K. Pamula
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Abstract
The present invention relates to providing improved hydrogen peroxide assays, as well as droplet actuators for conducting such assays. The droplet actuators of the invention may be used to conduct droplet-based hydrogen peroxide assays. They may also be associated with detectors for analyzing the results of the hydrogen peroxide assays of the invention. They may be provided as components of systems which control droplet operations and/or detection for conducting the hydrogen peroxide assays. Measurement by the detector may be used to quantify the presence of an analyte in a sample.
Claims32
A method of conducting a hydrogen peroxide assay for detecting an analyte, the method comprising:
- (a) combining hydrogen peroxide assay reagents and a sample into a droplet and conducting the assay in the droplet surrounded by oil in a droplet actuator to yield a detectable signal, the hydrogen peroxide reagents comprising an oxidase, a peroxidase, and a substrate which produces a signal molecule in the presence of a peroxidase and hydrogen peroxide, and the peroxidase provided in an amount effective to produce a substantially linear response, wherein the assay comprises a flash assay in which substantially all of the analyte is consumed, and wherein the sample and reagents for conducting the flash assay are surrounded by oil and are combined to form the droplet in the presence of a sensor sensing the signal, and wherein the combining is accomplished on a droplet actuator using electrowetting-mediated droplet operations;
- and
- (b) analyzing results of the assay based on measurement by the sensor.
A method of conducting a hydrogen peroxide assay, the method comprising:
- (a) combining in the presence of a sensor hydrogen peroxide assay reagents and a sample into a droplet and conducting the assay in the droplet surrounded by oil in a droplet actuator to yield a detectable signal, and wherein the reagents are selected to cause the assay to produce a substantially linear result, and wherein the sensor is sensing the signal upon the combining of the hydrogen peroxide assay reagents and a sample;
- and
- (b) analyzing results of the assay based on measurement by the sensor.
A method of conducting a hydrogen peroxide assay for detecting an analyte, the method comprising:
- (a) combining in the presence of a sensor hydrogen peroxide assay reagents and sample into a droplet and conducting the assay in the droplet surrounded by oil in a droplet actuator to yield a signal detectable by the sensor, wherein substantially all of the analyte is consumed in less than about 5 minutes, and wherein the sensor is sensing the signal upon the combining of the hydrogen peroxide assay reagents and sample;
- (b) detecting the signal;
- and
- (c) analyzing the signal to determine the quantity of analyte in the sample.
Dependencies are inferred from the claim text.