Sensitive and rapid determination of antimicrobial susceptibility

• US 9,841,422 B2
• Filed: 07/11/2014
• Issued: 12/12/2017
• Est. Priority Date: 07/12/2003
• Status: Active Grant

• Current Assignee: Accelerate Diagnostics, Inc.
• Original Assignee: Accelerate Diagnostics, Inc.
• Inventors: Goldberg, David A., Howson, David C., Metzger, Steven W., Buttry, Daniel A., Saavedra, Steven Scott
• Primary Examiner(s): LAM, ANN Y
• Application Number:
  US14/329,387
• Publication Number:
  US 20140323340A1
• Time in Patent Office: 1,250 Days
• Field of Search: None
• CPC Class Codes: B82Y 10/00 Nanotechnology for information processing, storage or transmission, e.g. quantum computing or single electron logic
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 20/00 Nanooptics, e.g. quantum optics or photonic crystals
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C12Q 1/18 Testing for antimicrobial activity of a material
• CPC Class Codes: G01N 33/5438 Electrodes
• CPC Class Codes: G01N 33/561 Immunoelectrophoresis
• CPC Class Codes: G01N 33/569 for microorganisms, e.g. protozoa, bacteria, viruses

Abstract

The present invention relates to moving microorganisms to a surface, where they are grown in the presence and absence of antimicrobials, and by monitoring the growth of the microorganisms over time in the two conditions, their susceptibility to the antimicrobials can be determined. The microorganisms can be moved to the surface through electrophoresis, centrifugation or filtration. When the movement involves electrophoresis, the presence of oxidizing and reducing reagents lowers the voltage at which electrophoretic force can be generated and allows a broader range of means by which the target can be detected. Monitoring can comprise optical detection, and most conveniently includes the detection of individual microorganisms. The microorganisms can be stained in order to give information about their response to antimicrobials.

Claims

1. A system for the identification of individual microorganisms in a sample comprising:

an enclosed chamber comprising;

a first electrode disposed on a detection surface, wherein the first electrode is a transparent conductive surface;

a second electrode disposed on a second surface;

an input port configured to transport a fluid into the chamber;

an output port configured to transport the fluid out of the chamber;

a capture surface disposed on the first electrode, wherein the capture surface comprises a binding agent configured to bind the individual microorganisms in spatially discrete locations;

an electrical controller operably linked to the first and second electrodes and configured to control potential between the first and second electrodes;

an optical detector configured to detect the individual microorganisms bound to the capture surface; and

a storage controller configured to perform analysis of an image obtained by the optical detector.

2. The system of claim 1, wherein the storage controller comprises a computer configured with software suitable for performing image analysis.

3. The system of claim 1, wherein the storage controller is configured to register the locations of individual microorganisms bound to the capture surface.

4. The system of claim 1, wherein the storage controller is configured to determine the morphology of individual microorganisms bound to the capture surface.

5. The system of claim 1, further comprising a first indicator configured to bind to a first microorganism.

6. The system of claim 5, further comprising a second indicator configured to bind to a second microorganism.

7. The system of claim 6, wherein the first indicator and the second indicator are fluorophores distinguishable by the detector by excitation wavelength, emission wavelength, or both excitation and emission wavelengths.

8. The system of claim 2, wherein the system is configured to perform detection and image analysis automatically.

9. The system of claim 2, wherein the first electrode comprises an indium tin oxide electrode.