Apparatus and method for detecting microscopic organisms using bacteriophage

• US 8,092,990 B2
• Filed: 03/31/2006
• Issued: 01/10/2012
• Est. Priority Date: 03/31/2005
• Status: Active Grant

• Current Assignee: Colorado School of Mines
• Original Assignee: Colorado School of Mines
• Inventors: Voorhees, Kent J.
• Primary Examiner(s): Steele, Amber D.
• Assistant Examiner(s): Shen, Bin
• Application Number:
  US11/394,936
• Publication Number:
  US 20070059725A1
• Time in Patent Office: 2,111 Days
• Field of Search: 435/5
• US Class Current: 435/5
• CPC Class Codes: C12Q 1/04 Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor C12Q1/6897 takes precedence
• CPC Class Codes: G01N 33/54388 based on lateral flow
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• CPC Class Codes: G01N 33/56911 Bacteria

Abstract

Methods and apparatus are provided for detection of microorganisms in a sample. Methods and apparatus of the invention are based on the specificity that phage, for example bacteriophage, have for target microorganisms, for example bacterium. Typically, phage adsorption to target microorganisms act as signal, or a signal target, for the presence of the target microorganism. Typically, the phage are labeled with a detectable signal. Apparatus of the invention are directed toward concentrating the phage adsorbed microorganisms at a predetermined site for flag dependent observation.

Claims

1. A method of determining the presence of a target microorganism in a sample, the method comprising:

combining with the sample an amount of flagged phage capable of binding to an outside surface of the target microorganism to create a flag labeled target microorganism;

providing conditions and an amount of time sufficient to allow the flagged phage to bind to the outside surface of the target microorganism; and

assaying the phage exposed sample to detect the presence of the flag labeled microorganism, without detecting phage amplified within the target microorganism and, wherein presence of the flag bound to the outside surface of the target microorganism indicates presence of the microorganism,

wherein the detecting the presence of the flag labeled microorganism is by concentrating the flag labeled microorganism to enhance the capacity to detect the flag labeled microorganisms, and

wherein the concentrating of flag labeled microorganisms is on a flow strip device having one or more immobilization zones, wherein the flag labeled microorganisms are captured and concentrated within the one or more immobilization zones.

2. The method of claim 1 wherein the flagged phage is a phage labeled with colorimetric particles.

3. The method of claim 2 wherein the colorimetric particles are colloidal gold particles.

4. The method of claim 1 wherein the flagged phage is a phage labeled with a fluorescent tag.