John Baker Wheeler

US Patent:

8216780, Jul 10, 2012

Filed:

Jun 2, 2009

Appl. No.:

12/476796

Inventors:

Jonathan Drew Smith - Boulder CO, US
Breanna Leigh Dreiling - Longmont CO, US
Breanna Christine Smith - Lyons CO, US
John H. Wheeler - Boulder CO, US

Assignee:

MicroPhage (TM) Incorporated - Longmont CO

International Classification:

C12Q 1/70
C12Q 1/02

US Classification:

435 5, 435 29

Abstract:

A method of determining the presence or absence of a target microorganism in a sample to be tested, the method comprising: (a) combining with the sample an amount of bacteriophage capable of attaching to the target microorganism to create a bacteriophage-exposed sample; (b) providing conditions to the bacteriophage-exposed sample sufficient to allow the bacteriophage to attach to the target microorganism while inhibiting phage replication in a potentially cross-reactive, non-target microorganism; and (c) assaying the bacteriophage-exposed sample to detect the presence or absence of a bacteriophage marker to determine the presence or absence of the target microorganism; wherein the amount of the bacteriophage is between 10% to 70% of the threshold number of bacteriophage that the assay can detect, or between 1×10pfu/mL and 7×10pfu/mL.

US Patent:

20050003346, Jan 6, 2005

Filed:

Apr 12, 2004

Appl. No.:

10/823294

Inventors:

Kent Voorhees - Golden CO, US
John Rees - Golden CO, US
John Wheeler - Boulder CO, US
Angelo Madonna - Tooale UT, US

Assignee:

Colorado School Of Mines - Golden CO

International Classification:

C12Q001/70

US Classification:

435005000

Abstract:

A method for detecting one or more target bacteria in a raw sample where: 1) bacteriophage(s) specific to each target bacterium are added to the raw sample, 2) the test sample is incubated, and 3) the test sample is tested for the presence of each phage in sufficient numbers to indicate the presence of the associated target bacteria in the raw sample. In one embodiment, each phage is initially added to the raw sample in concentrations below the detection limit of the final phage detection process. In another embodiment, the parent phages are tagged in such a way that they can be separated from the progeny phage prior to the detection process. Preferred phage detection processes are immunoassay methods utilizing antibodies that bind specifically to each phage. Antibodies can be used that bind to the protein capsid of the phage. Alternatively, the phage can by dissociated after the incubation process and the sample tested for the presence of individual capsid proteins or phage nucleic acids. The invention can be used to test target bacteria for antibiotic resistance.

US Patent:

20050250096, Nov 10, 2005

Filed:

Jun 7, 2005

Appl. No.:

11/147800

Inventors:

John Wheeler - Boulder CO, US
Jon Rees - Golden CO, US
G. Gaisford - Denver CO, US

Assignee:

Microphage, Incorporated - Longmont CO

International Classification:

C12Q001/70
C12Q001/04
C12Q001/68

US Classification:

435005000, 435034000, 435006000

Abstract:

A method of detecting the presence or absence of a target microorganism in a sample to be tested comprising: combining with the sample, bacteriophage capable of infecting the target microorganism to create a bacteriophage exposed sample; providing conditions to the bacteriophage exposed sample sufficient to allow the bacteriophage to infect the target microorganism to create injected bacteriophage nucleic acid, additional bacteriophage nucleic acid, intermediate bacteriophage protein, or additional bacteriophage protein; and assaying the nucleic acid or protein to determine the presence or absence of the target microorganism. The microorganism is preferably dissociated or lysed prior to the completion of the bacteriophage replication process. The sample is an insitu sample and the microorganism is isolated from the insitu matrix. The assaying comprises comparing the test nucleic acid or protein level to a reference level.

US Patent:

20070178450, Aug 2, 2007

Filed:

Jan 25, 2007

Appl. No.:

11/698673

Inventors:

John H. Wheeler - Boulder CO, US
Jon C. Rees - Longmont CO, US
Gregory S. Gaisford - Denver CO, US

Assignee:

MicroPhage (TM) Incorporation - Longmont CO

International Classification:

C12Q 1/70
C12Q 1/68

US Classification:

435 5, 435 6

Abstract:

A predetermined amount of parent bacteriophage capable of infecting a target microorganism is added to a sample to create a bacteriophage-exposed sample; the sample is incubated for a defined incubation time and assayed to determine the level of a bacteriophage or bacterial marker in the sample; and if the measured marker level has increased, then the initial concentration of the microorganism exceeds a specific threshold value. An antibiotic in different concentrations is added to different and separate portions of the sample and tested to determine if the bacteriophage marker is present and thereby determine the Minimum Inhibitory Concentration (MIC) of a given antibiotic. The antibiotic preferably is an antibiotic that inhibits DNA replication or protein synthesis.

US Patent:

20080286757, Nov 20, 2008

Filed:

Sep 15, 2006

Appl. No.:

12/066806

Inventors:

G. Scott Gaisford - Denver CO, US
John H. Wheeler - Boulder CO, US
Jon C. Rees - Longmont CO, US
Scott D. Conlin - Boulder CO, US

Assignee:

MicroPhage Incorporated - Longmont CO

International Classification:

C12Q 1/70

US Classification:

435 5

Abstract:

A sample is tested for the presence of bacteria, such as in an automatic blood culturing apparatus. If bacteria are determined to be present, a bacteriophage-based bacteria identification process is performed to identify the bacteria present. A plurality of bacteria detection processes, such as a blood culture test and Gram stain test may be carried out prior to the bacteria identification process. A bacteriophage-based antibiotic resistance test or antibiotic susceptibility test is also conducted on the sample.

US Patent:

20090246752, Oct 1, 2009

Filed:

Dec 30, 2008

Appl. No.:

12/346656

Inventors:

Kent Voorhees - Golden CO, US
Jon Rees - Longmont CO, US
John H. Wheeler - Boulder CO, US
Angelo Madonna - Tooele UT, US

Assignee:

MicroPhage TM Incorporated - Longmont CO

International Classification:

C12Q 1/70

US Classification:

435 5

Abstract:

A method for detecting one or more target bacteria in a raw sample where: 1) bacteriophage(s) specific to each target bacterium are added to the raw sample, 2) the test sample is incubated, and 3) the test sample is tested for the presence of each phage in sufficient numbers to indicate the presence of the associated target bacteria in the raw sample. In one embodiment, each phage is initially added to the raw sample in concentrations below the detection limit of the final phage detection process. In another embodiment, the parent phages are tagged in such a way that they can be separated from the progeny phage prior to the detection process. Preferred phage detection processes are immunoassay methods utilizing antibodies that bind specifically to each phage. Antibodies can be used that bind to the protein capsid of the phage. Alternatively, the phage can by dissociated after the incubation process and the sample tested for the presence of individual capsid proteins or phage nucleic acids. The invention can be used to test target bacteria for antibiotic resistance.

US Patent:

20090286225, Nov 19, 2009

Filed:

Mar 11, 2009

Appl. No.:

12/402337

Inventors:

John H. Wheeler - Boulder CO, US
Michael Fiechtner - Poway CA, US
Jonathan D. Smith - Boulder CO, US
Duane Bush - Fort Collins CO, US
Alene A. Campbell - Boulder CO, US
Breanna C. Smith - Lyons CO, US

Assignee:

MicroPhageTM Incorporated - Longmont CO

International Classification:

C12Q 1/70
C12M 1/00

US Classification:

435 5, 4352877, 4352872, 4353031

Abstract:

Bacteriophage are combined with a test sample in an incubator, and the bacteriophage-exposed test sample is conjugated and applied to a sample pad in contact with a lateral flow strip to determine the presence or absence of a target bacterium. The conjugation may be performed in the sample pad or prior to application of the bacteriophage-exposed test sample to the pad. The incubator comprises a bacteriophage container and an incubation container separated by a valve. The test sample may be inserted into the incubation chamber using a swab or a rod with a piercing tip and a sample collection eye. The valve comprises a breakable stem. An antibiotic may be added to the test sample to determine the antibiotic resistance or susceptibility of the bacterium.