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US 12,571,730 B2Granted
Label-free bacterial detection
AssigneeMiami University
InventorsKevin Yehl, Patrick Needham
Life of this patent
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Abstract
Disclosed herein are systems and methods for quantifying bacteriophage virulence, measuring phage-host dynamics and parameters, including phage-host range, phage interactions with biological samples and with immune systems, and label-free bacterial detection/diagnostics, that are amenable to automation, high-throughput, and functional in complex media. In some embodiments, a label-free interferometry system transduces the light reflected by a sensor and any molecules attached thereto to a real-time signal comprising a sensorgram from which infectivity parameters such as binding kinetics and lysis time can be derived.
Claims20
A method of measuring phage-host dynamics and parameters using bacteriophage infection of a bacteria comprising:
- providing a sensor with a tip;
- attaching and immobilizing to the sensor a plurality of bacteriophages having a phage-host range to obtain a functionalized sensor;
- irradiating the functionalized sensor with light and detecting and obtaining in real-time an at least one wavelength of white light reflected by the tip of the functionalized sensor with the bacteriophages attached thereto and corresponding to a baseline interference pattern measurement;
- contacting the functionalized sensor with a capture medium comprising a liquid mixture further comprising an at least one host;
- irradiating the functionalized sensor with light, such that if the bacteriophages bind to the at least one host, the binding of the bacteriophages to the at least one host shifts the baseline interference pattern measurement;
- detecting and obtaining in real-time a modified interference pattern measurement, wherein the modified interference pattern measurement comprises a measurement of the at least one wavelength of white light reflected from the tip of the functionalized sensor as shifted by the bacteriophages and the at least one host attached thereto;
- comparing the modified interference pattern measurement to the baseline interference pattern measurement to produce a sensorgram;
- wherein a binding signal and a lysis signal are detected, wherein the binding signal comprises an amount of the at least one host bound to the bacteriophages and the lysis signal comprises an amount of the at least one host bound to the bacteriophages while the at least one host is being lysed by the bacteriophages;
- screening the phage-host range from the sensorgram.
6 more elements
A system for measuring phage-host dynamics and parameters using bacteriophage infection of a bacteria comprising:
- a sensor having a tip;
- a host attached and immobilized to the sensor tip to form a functionalized sensor;
- wherein when the functionalized sensor is irradiated with light, an at least one wavelength of white light is reflected by the tip of the functionalized sensor with the host attached thereto, corresponding to a baseline interference pattern measurement;
- a capture medium comprising a liquid mixture of bacteriophages provided to the functionalized sensor, the bacteriophages having a phage-host range;
- wherein if the host is within the phage-host range of the bacteriophages, the bacteriophages bind to the host;
- wherein the binding of the bacteriophages to the host shifts the baseline interference pattern measurement to form a modified interference pattern measurement, wherein the modified interference pattern measurement comprises a measurement of the at least one wavelength of white light reflected from the tip of the functionalized sensor with as shifted by the host and the bacteriophages attached thereto;
- a comparison of the modified interference pattern measurement to the baseline interference pattern measurement, the comparison comprising a sensorgram;
- wherein, if the host is within the phage-host range of the bacteriophages, a binding signal and a lysis signal are detected, wherein the binding signal comprises an amount of the at least one host bound to the bacteriophages and the lysis signal comprises an amount of the at least one host bound to the bacteriophages while the at least one host are is being lysed by the bacteriophages, and the phage-host range is screened.
5 more elements
A method for label-free bacterial detection using bacteriophage infection of a bacteria comprising:
- providing a sensor with a tip;
- attaching and immobilizing to the sensor a plurality of bacteriophages having a phage-host range to obtain a functionalized sensor;
- irradiating the functionalized sensor with light and detecting and obtaining in real-time an at least one wavelength of white light reflected by the tip of the functionalized sensor with the bacteriophages attached thereto and corresponding to a baseline interference pattern measurement;
- contacting the functionalized sensor with a capture medium comprising a liquid mixture;
- irradiating the functionalized sensor with light, such that if the capture medium comprises an at least one host within the phage-host range of the bacteriophages, the bacteriophages bind to the at least one host, and the binding of the bacteriophages to the at least one host shifts the baseline interference pattern measurement;
- detecting and obtaining in real-time a modified interference pattern measurement, wherein the modified interference pattern measurement comprises a measurement of the at least one wavelength of white light reflected from the tip of the functionalized sensor as shifted by the bacteriophages and the at least one host attached thereto;
- comparing the modified interference pattern measurement to the baseline interference pattern measurement to produce a sensorgram;
- wherein, in the presence of at least one host within the phage-host range of the bacteriophages, a binding signal and a lysis signal are detected, wherein the binding signal comprises an amount of the at least one host bound to the bacteriophages and the lysis signal comprises an amount of the at least one host bound to the bacteriophages while the at least one host is being lysed by the bacteriophages.
5 more elements
Dependencies are inferred from the claim text.