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US 2016/0252502 A1Published application
Detection assays and methods
AssigneeRegents of the University of Minnesota
InventorAbdennour Abbas
Life of this patent
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Abstract
This disclosure describes assay methods and kits for detecting a target. The methods and kits can be used to detect a target that is present in a sample at low concentration because the methods and kits amplify the signal indicating the presence of target in the sample. Generally, the methods and kits involve nanoparticle aggregation as a detectable signal that is enhanced by a trigger released from a vesicular compartment when the target is bound to a capture agent.
Claims33
A method for detecting a target, the method comprising:
- providing a capture agent that specifically binds to the target;
- contacting a sample that includes the target with the capture agent, thereby allowing the target to bind to the capture agent;
- removing unbound sample;
- providing a recognition unit that specifically binds to the target and comprises a vesicular compartment that contains a chromogenic trigger;
- contacting the recognition unit with the target bound to the capture agent, thereby allowing the recognition unit to specifically bind to the target;
- removing unbound recognition units;
- contacting a lysis agent with the vesicular compartment, thereby allowing the lysis agent to lyse the vesicular compartment, releasing the chromogenic trigger;
- and
- contacting a composition that comprises a chromogenic reagent with the released chromogenic trigger, thereby allowing the chromogenic trigger to induce the chromogenic reagent to produce a colorimetric signal.
6 more elements
A method for detecting a target, the method comprising:
- providing a capture agent that specifically binds to the target;
- contacting a sample that includes the target with the capture agent, thereby allowing the target to bind to the capture agent;
- removing unbound sample;
- providing a recognition unit that specifically binds to the target and comprises a primary vesicular compartment that contains an activatable lysis agent;
- contacting the recognition unit with the target bound to the capture agent, thereby allowing the recognition unit to specifically bind to the target;
- removing unbound recognition units;
- contacting the primary vesicular compartment with a activation agent that activates the lysis agent, thereby activating the lysis agent to lyse the primary vesicular compartment and release the lysis agent;
- providing a composition comprising secondary vesicular compartments containing a chromogenic trigger;
- contacting the secondary vesicular compartments with the lysis agent released from the primary vesicular compartment, thereby allowing the lysis agent to lyse at least a portion of the secondary vesicular compartments, releasing the chromogenic trigger;
- and
- contacting a composition that comprises a chromogenic reagent with the released chromogenic trigger, thereby allowing the chromogenic trigger to induce the chromogenic reagent to produce a colorimetric signal.
8 more elements
A method for detecting a target, the method comprising:
- providing a capture agent that specifically binds to the target;
- contacting a sample that includes the target with the capture agent, thereby allowing the target to bind to the capture agent;
- removing unbound sample;
- providing a recognition unit that specifically binds to the target and comprises a lysis agent;
- contacting the recognition unit with the target bound to the capture agent, thereby allowing the recognition unit to specifically bind to the target;
- removing unbound recognition units;
- providing a composition comprising vesicular compartments containing a chromogenic trigger;
- contacting the vesicular compartment composition with the lysis agent, thereby allowing the lysis agent to lyse at least a portion of the vesicular compartments, releasing the chromogenic trigger;
- and
- contacting a composition that comprises a chromogenic reagent with the released chromogenic trigger, thereby allowing the chromogenic trigger to induce the chromogenic reagent to produce a colorimetric signal.
7 more elements
A kit for detecting a target, the kit comprising:
- a recognition unit comprising:
- a recognition moiety that specifically binds to the target, and
- a signal initiation moiety that comprises:
- a lysis agent,
- a vesicular compartment that contains a chromogenic trigger, or
- a vesicular compartment that contains an activatable lysis agent, or
- a vesicular compartment that includes a compound that can generate a detectable signal;
- and
- a chromogenic reagent.
6 more elements
A method for detecting a target, the method comprising:
- providing a capture agent that specifically binds to the target, the capture agent displaceably bound to a lysis agent;
- contacting a sample that includes the target with the capture agent, thereby allowing the target to bind to the capture agent and displace the lysis agent;
- contacting a composition comprising vesicular compartments containing a chromogenic trigger with the displaced lysis agent, thereby allowing the displaced lysis agent to lyse at least a portion of the vesicular compartments, releasing the chromogenic trigger;
- and
- contacting a composition that comprises a chromogenic reagent with the released chromogenic trigger, thereby allowing the chromogenic trigger to induce the chromogenic reagent to produce a colorimetric signal.
2 more elements
Dependencies are inferred from the claim text.