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US 11,686,729 B2Granted
Bacteriophage-based electrochemical bacterial sensors, systems, and methods
AssigneeUniversity of Georgia Research Foundation, Inc.
InventorsRamaraja P. Ramasamy, Yan Zhou
Life of this patent
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Abstract
The present disclosure includes methods and systems of detecting bacteria in a sample using phage-functionalized sensors, methods of enriching a sample with phage-functionalized magnetic particles, phage-functionalized magnetic particles and methods of making phage-functionalized magnetic particles.
Claims22
A system for detecting bacteria in a sample, the method system comprising:
- magnetic particles functionalized with a first plurality of bacteriophages specific for a target strain of bacteria to be detected, wherein the bacteriophages are immobilized to the magnetic particle such that a majority of the phage are oriented in a head-in configuration relative to the magnetic particle such that a head of the bacteriophage is coupled to the magnetic particle and the tail of the bacteriophage is free to attach to bacterial cells of the target strain in a sample containing the target bacteria;
- a magnet configured to provide a magnetic field to the sample and to separate magnetic particle-coupled bacterial cells from other components in the sample to provide an enriched sample comprising a greater concentration of the target bacteria than the original sample;
- a bacteriophage-modified electrode comprising:
- an electrode material,
- a layer of multi-walled, carbon nanotubes (MWCNTs) on a surface of the electrode material thereby providing a MWCNT-modified electrode, wherein the MWCNT-modified electrode further comprises:
- an applied positive potential sufficient to provide a net positive surface charge to the MWCNT-modified electrode, and
- a second plurality of bacteriophages specific for the target strain of bacteria, the second plurality of bacteriophages immobilized to the MWCNTs via a linker, wherein the net positive surface charge of the MWCNT modified electrode is sufficient to facilitate charge-directed immobilization of the second plurality of bacteriophages such that the majority of the second plurality of bacteriophages on the electrode are oriented in a head-in, tail-out configuration relative to the electrode surface such that a head of the bacteriophage is coupled to the multi-walled, carbon nanotubes via the linker, wherein the second plurality of bacteriophages are capable of binding the target bacteria in the enriched sample;
- and
- a three-electrode electrochemical cell, in which the bacteriophage-modified electrode is a working electrode, wherein the system is configured to produce a detectable impedimetric signal upon binding of target bacteria to the phage-modified electrode.
6 more elements
A system for detecting bacteria in a sample, the method comprising:
- magnetic particles functionalized with a first plurality of bacteriophages specific for a target strain of bacteria to be detected, wherein the bacteriophages are immobilized to the magnetic particle such that a majority of the phage are oriented in a head-in configuration relative to the magnetic particle such that a head of the bacteriophage is coupled to the magnetic particle and the tail of the bacteriophage is free to attach to bacterial cells of the target strain in a sample containing the target bacteria;
- a magnet configured to provide a magnetic field to the sample and to separate magnetic particle-coupled bacterial cells from other components in the sample to provide an enriched sample comprising a greater concentration of the target bacteria than the original sample;
- a bacteriophage-modified electrode comprising:
- an electrode material,
- a layer of multi-walled, carbon nanotubes (MWCNTs) on a surface of the electrode material thereby providing a MWCNT-modified electrode, wherein the MWCNT-modified electrode further comprises:
- a cationic polymer and an applied positive potential, such that a combined positive charge from the cationic polymer and the applied positive potential provides a net positive surface charge, and
- a second plurality of bacteriophages specific for the target strain of bacteria, the second plurality of bacteriophages immobilized to the MWCNTs via a linker, wherein the net positive surface charge of the MWCNT modified electrode is sufficient to facilitate charge-directed immobilization of the second plurality of bacteriophages to the electrode surface via the linker in a head-in, tail out configuration relative to the electrode surface such that a head of the bacteriophage is coupled to the multi-walled, carbon nanotubes via the linker, wherein the second plurality of bacteriophages are capable of binding the target bacteria in the enriched sample;
- and
- a three-electrode electrochemical cell, in which the bacteriophage-modified electrode is a working electrode, wherein the system is configured to produce a detectable impedimetric signal upon binding of target bacteria to the phage-modified electrode.
6 more elements
Dependencies are inferred from the claim text.