Bacteriophage-based biosensor for microbial detection

Patent metadata

Field	Value
Publication number	US-9921219-B2
Application number	US-201514837621-A
Country	US
Kind code	B2
Filing date	Aug 27, 2015
Priority date	Aug 27, 2014
Publication date	Mar 20, 2018
Grant date	Mar 20, 2018

Abstract

A biosensor for detecting a microorganism in a sample includes a polymer matrix and an immobilized layer of bacteriophages on the surface of the polymer matrix, wherein interaction of the bacteriophage with the microorganism causes a detectable physicochemical change in the polymer matrix. A method of detecting a microorganism in a sample using the biosensor includes the steps of contacting the sample with the biosensor; allowing the bacteriophage to bind to the microorganism in the sample; and detecting a physicochemical change in the polymer matrix, the change being indicative of the presence of the microorganism.

First claim

What is claimed is:

1. A biosensor for detecting a microorganism in a sample, the biosensor comprising a polymer matrix and an immobilized layer of bacteriophages on the surface of the polymer matrix, wherein the bacteriophages comprise a phage carrying a reporter gene that codes for a depolymerase that is expressed when the phage infects the microorganism and the polymer matrix comprises a material detectably responsive to the depolymerase by degradation of the material.

2. The biosensor of claim 1 , wherein the bacteriophage comprises a native or genetically modified phage.

3. The biosensor of claim 1 , wherein the depolymerase is a polyhydroxyalkanoate (PHA) depolymerase.

4. The biosensor of claim 1 , wherein the polymer matrix comprises at least one polymer, a blend of polymers, a co-polymer, or a composite material.

5. The biosensor of claim 4 , wherein the composite comprises a degradable polymer and a non-degradable polymer.

6. The biosensor of claim 4 , wherein the polymer matrix defines an optical grating.

7. The biosensor of claim 4 , wherein the polymer matrix comprises a porous material.

8. The biosensor of claim 4 , wherein the polymer matrix comprises a material that becomes porous upon exposure to a degradation-inducing substance.

9. The biosensor of claim 4 , wherein the polymer matrix is loaded with a dye.

10. The biosensor of claim 4 , wherein the polymer matrix is loaded with a plurality of conducting particles, or formed as a substrate for a radio frequency identification (RFID) antenna, or a coating for an antenna of a RFID tag.

11. The biosensor of claim 4 , where the polymer matrix is patterned at the nano-, micro- or millimeter-scale, or a combination thereof, the polymer matrix being a pure polymer, a co-polymer, or a polymer blended with a non-degradable polymer or non-polymeric material.

12. The biosensor of claim 1 , wherein the degradable material is selected from polylactic acid, polyhydroxyalkanoate, polycaprolactone.

13. The biosensor of claim 4 , wherein the polymer matrix is in the form of a free-standing film or coating on a carrier layer.

14. A method of detecting a microorganism in a sample, comprising the steps of:
(a) contacting the sample with the biosensor of claim 1 ;
(b) allowing bacteriophage to bind to the microorganism in the sample; and
(c) detecting a physicochemical change in the polymer matrix caused by interaction of the bacteriophage with the microorganism.

Assignees

• Univ Alberta

Inventors

• Elias Anastasia
• Sauvageau Dominic
• Storms Zachary
• Wang Chanchan
• Anbukarasu Preetam
• Martinez-Tobon Diana

Classifications

• G01N33/54373

  involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings · CPC title

• G01N33/56911 Primary

  Bacteria · CPC title