Method and apparatus for the detection of pathogens, parasites, toxins and desired chemical compounds

• US 8,652,311 B1
• Filed: 04/01/2011
• Issued: 02/18/2014
• Est. Priority Date: 10/02/2006
• Status: Active Grant

• Current Assignee: Ebtisam Wilkins
• Original Assignee: Ebtisam Wilkins
• Inventors: Wilkins, Ebtisam
• Primary Examiner(s): Noguerola, Alex
• Application Number:
  US13/065,926
• Time in Patent Office: 1,054 Days
• Field of Search: 204/403.01, 204/403.02, 204409-412, 205/777.5, 205/778, 205/792
• US Class Current: 204/403.01
• CPC Class Codes: G01N 33/5438 Electrodes

Abstract

The present invention is directed to a method and apparatus for an immunoassay technique that uses amperometric measurements to rapidly analyze different pathogenic microorganisms, including bacteria, viruses, toxins, and parasites. A conductive membrane is used to provide support for antibody immobilization and serve as a working electrode. The proposed technique is adaptable for use with different materials so as to form a membrane having a pore size that is suited to the particular application. A compact and simple disposable element can be used. The immunoassay can be automated using microprocessor control so as to reduce the amount of human intervention in sample analysis.

Claims

1. A flow-through immuno-sensor with electrochemical detection capabilities, comprising:

a body;

an electrode assembly housed within the body, the electrode assembly comprising

a first substrate supporting a working electrode

a second substrate supporting a reference electrode and a counter electrode;

a detector configured to detect an electrical signal generated by the working electrode;

wherein the body also has a fluid inlet configured to introduce a fluid sample into the body;

a fluid path in communication with the fluid inlet, the fluid path being configured to allow fluid to encounter the working electrode, the counter electrode, and the reference electrode; and

an outlet channel;

wherein the working electrode comprises immobilized antibody or antigen and is configured for an enzyme reaction so forming a reaction region; and

wherein when test sample comprising a target analyte antibody or antigen is introduced to the reaction region via the inlet channel under sufficient conditions for the target analyte to bind to the immobilized antibody or antigen, unbound molecules in test sample exit the reaction region via the outlet channel, and thereafter when one or more detection molecules are introduced to the reaction region an enzymatic reaction will occur, the product of which is able to produce a detectable alteration in the working electrode'"'"'s signal output.

2. The flow-through immuno-sensor with electrochemical detection capabilities of claim 1, further comprising an immuno-selective membrane in fluidic communication with the fluid inlet and the working electrode.

3. The flow-through immuno-sensor with electrochemical detection capabilities of claim 1, wherein the working electrode is screen-printed on the first substrate.

4. The flow-through immuno-sensor with electrochemical detection capabilities of claim 1 wherein the antibody or antigen enables physical contact between the target analyte and the working electrode.

5. The flow-through immuno-sensor with electrochemical detection capabilities of claim 4 wherein the test sample is selected from the group consisting of blood, body fluid, animal fluid, environmental sample, and warfare agent.

6. The flow-through immuno-sensor with electrochemical detection capabilities of claim 4 wherein the working electrode comprises a coating of electro-conductive layer including conductive particles, wherein the conductive particles are carbon particles, graphite, or ULTI immuno-sorbent;

and wherein the coating further comprises a multi analyte-specific antibody or antigen.

7. The flow-through immuno-sensor with electrochemical detection capabilities of claim 1 wherein the working electrode comprises multi micro-channels and wherein the micro-channels are fluidly connected to the fluid inlet such that the fluid sample flows through the channels.

8. A multi-channel flow-through sensor with electrochemical detection capabilities, comprising:

a substantially planar body;

multiple fluid channels connected to the body, each fluid channel in fluidic communication with an electrode assembly, each electrode assembly comprising a substrate and a working electrode, along with a fluid inlet configured to introduce a fluid sample into the body from a fluid channel and to present a target analyte in the fluid sample to the working electrode such that an electrical signal generated by the working electrode is altered; and

a detector configured to detect the electrical signal generated by the working electrode;

wherein antibody or antigen is immobilized on the carbon particles deposited on top of the working electrode that enables physical contact between the target analyte and the working electrode.

9. A multi-channel flow-through sensor with electrochemical detection capabilities the sensor of claim 8 wherein the antibody or antigen produces a chemical reaction when exposed to the target analyte and the product of the chemical reaction alters the electrical signal generated by the working electrode.