DEVICES AND METHODS FOR TARGET ANALYTE DETECTION IN LIQUID SAMPLES

• US 20170233785A1
• Filed: 10/16/2015
• Published: 08/17/2017
• Est. Priority Date: 10/16/2014
• Status: Active Grant

• Current Assignee: The General Hospital Corporation (Mass General Brigham, Inc.)
• Original Assignee: The General Hospital Corporation (Mass General Brigham, Inc.)
• Inventors: Palanisami, Akilan, Hasan, Tayyaba, Khan, Shazia
• Granted Patent: US 10,465,226 B2
• Time in Patent Office: Days
• CPC Class Codes: B01L 2300/042 Caps; Plugs
• CPC Class Codes: B01L 2300/044 pierceable, e.g. films, membranes
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0874 Three dimensional network
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/12 Specific details about materials
• CPC Class Codes: B01L 2400/049 vacuum
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502746 characterised by the means for controlling flow resistance, e.g. flow controllers, baffles B01L3/502738 takes precedence
• CPC Class Codes: C12Q 1/14 Streptococcus; Staphylococcus
• CPC Class Codes: G01N 33/56938 Staphylococcus
• CPC Class Codes: G01N 33/582 with fluorescent label

Abstract

One aspect of the present disclosure relates to a device for detecting a target analyte in a liquid sample. The device can comprise a housing. The housing can include an inlet for receiving a liquid sample, an outlet for removing a volume of the liquid sample from the device, a filter associated with the outlet and being sized and dimensioned to retain a target analyte on a surface thereof, and a flow system comprising at least one channel that is in communication with the inlet and the outlet. At least a portion of the at least one channel can be located substantially adjacent the surface of the filter and be shaped and dimensioned to reduce the amount of unreacted fluorescent probe available to create the background interference during detection of the target analyte.

Claims

1. A device for detecting a target analyte in a liquid sample, the device comprising:

a housing that includes;

an inlet for receiving a liquid sample;

an outlet for removing a volume of the liquid sample from the device;

a filter associated with the outlet and being sized and dimensioned to retain a target analyte on a surface thereof; and

a flow system comprising at least one channel that is in communication with the inlet and the outlet, at least a portion of the at least one channel being located substantially adjacent the surface of the filter and shaped and dimensioned to reduce the amount of unreacted fluorescent probe available to create background fluorescence during detection of the target analyte.

2. The device of claim 1, wherein a diameter of the portion is less than about 50 microns.

3. The device of claim 1, further including a release mechanism coupled to the housing and in communication with the at least one channel, the release mechanism being configured to permit selective release of the retained target analyte off of the surface of the filter for detection of the target analyte.

4. The device of claim 3, wherein the release mechanism includes a plug member that is slidably disposed within a second outlet of the housing, the second outlet being in fluid communication with the at least one channel.

5. The device of claim 3, wherein the release mechanism is a syringe fluidly coupled to the second outlet.

6. The device of claim 1, wherein the target analyte is a pathogen or biomolecule associated therewith.

7. The device of claim 1, being configured as a multi-channel device and further comprising:

a second inlet for receiving a second liquid sample;

a second outlet for removing a volume of the second liquid sample from the device;

a second filter associated with the second outlet and being sized and dimensioned to retain a target analyte on a surface thereof; and

a second flow system comprising at least one channel that is in communication with the second inlet and the second outlet.

8. The device of claim 7, wherein the second inlet, the second outlet, the second filter, and the second flow system are arranged about the device in parallel to the inlet, the outlet, the filter, and the flow system.

9. The device of claim 7, further including a second release mechanism coupled to the housing and in communication with the at least one channel of the second flow system.

10. The device of claim 1, wherein the at least one channel is partitioned into a first channel and a second channel by a filter having oppositely disposed first and second surfaces, the first surface of the filter being adapted to collect and concentrate the target analyte when a liquid sample containing the target analyte is flowed through the first and second channels.

11. The device of claim 1, wherein at least one of the filter, a wall defining the inlet, and a wall defining the flow channel is partially or entirely coated with the fluorescent probe.

12. A method for detecting a target analyte in a liquid sample, the method comprising the steps of:

introducing, through an inlet of a device, a liquid sample either before, during, or after introduction of a detection reagent through the inlet, the device including a flow system comprising at least one channel that is in communication with the inlet and an outlet, at least a portion of the at least one channel being located substantially adjacent the surface of the filter and shaped and dimensioned to reduce the amount of unreacted fluorescent probe available to create the background fluorescence;

removing, from an outlet of the device, a volume of the liquid sample to cause a target analyte, if present, to be retained on a surface of a filter associated with the outlet;

exposing the target analyte to a detection reagent specific to the target analyte; and

determining the presence of the target analyte in the liquid sample.

13. The method of claim 12, wherein the target analyte is a target pathogen or biomolecule associated therewith.

14. The method of claim 12, wherein the detection reagent is a fluorescent probe specific to the target pathogen or biomolecule associated therewith.

15. The method of claim 14, wherein the fluorescent probe is beta-Lactamase Enzyme Activated Fluorophore (β

-LEAF) and the target pathogen is S. aureus.

16. The method of claim 12, wherein the target analyte, if present, is concentrated on the surface of the filter associated with the outlet such that the concentration is greater than the concentration of the target analyte in the liquid sample.

17. A method for detecting a target pathogen, or biomolecule associated therewith, in a liquid sample, the method comprising the steps of:

introducing, through an inlet of a device, a liquid sample either before, during, or after introduction of a fluorescent probe specific to the target pathogen, or biomolecule associated therewith, through the inlet;

removing, from an outlet of the device, a volume of the liquid sample to cause a target pathogen, or biomolecule associated therewith, if present, to be retained on a surface of a filter associated with the outlet;

actuating a release mechanism of the device to cause release of the retained target pathogen, or biomolecule associated therewith, off of the surface of the filter; and

determining, by fluoroscopy, the presence of the target pathogen, or biomolecule associated therewith, in the liquid sample;

whereby release of the target pathogen, or biomolecule associated therewith, reduces background fluorescence associated with the fluorescent probe during the determining step.

18. The method of claim 17, wherein the fluorescent probe is β

-LEAF and the target pathogen is S. aureus.

19. The method of claim 17, wherein the target pathogen, if present, is concentrated on the surface of the filter associated with the outlet such that the concentration is greater than the concentration of the target pathogen in the liquid sample.

20. The method of claim 17, wherein the step of actuating a release mechanism further includes advancing a plug member, which is slidably disposed in a second outlet of the housing, axially away from the housing so that the liquid sample flows through the flow system towards the second outlet.