MULTIPLEXED INSTRUMENT-FREE BAR-CHART SPINCHIP INTEGRATED WITH NANOPARTICLE-MEDIATED APTASENSORS FOR VISUAL QUANTITATIVE DETECTION OF MULTIPLE PATHOGENS

• US 20200132675A1
• Filed: 07/17/2019
• Published: 04/30/2020
• Est. Priority Date: 07/17/2018
• Status: Active Application

• Current Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Inventors: Li, XiuJun, Dominguez, Delfina, Wei, Xiafeng
• Application Number:
  US16/514,521
• Publication Number:
  US 20200132675A1
• Time in Patent Office: Days
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: C12Q 1/6813 Hybridisation assays
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 1/6832 Enhancement of hybridisation reaction
• CPC Class Codes: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: G01N 33/5091 for testing the pathological state of an organism
• CPC Class Codes: G01N 33/54326 Magnetic particles
• CPC Class Codes: G01N 33/54346 Nanoparticles
• CPC Class Codes: G01N 33/553 Metal or metal coated
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• CPC Class Codes: G01N 33/56911 Bacteria

Abstract

A point-of-care testing (POCT) quantitative pathogen detection device is provided, without the aid of any detectors. In an illustrative embodiment, a POCT pathogen detection device includes an inlet microwell for receiving a substance, the inlet microwell connected to a distribution channel to distribute the substance to an analyzing component. The device also includes an analyzing component. that includes a first pathogen detection component. The first pathogen detection component includes a platinum nanoparticle-labeled magnetic DNA-probe configured to propel a dye through a bar-chart channel when a pathogen in the substance reacts platinum nanoparticle-labeled magnetic DNA-probe. The platinum nanoparticle-labeled magnetic DNA-probe is configured to react with a first pathogen. The device also includes at least one magnet configured to keep unreacted platinum nanoparticle-labeled magnetic DNA-probe in a sample recognition microwell, thereby inhibiting propulsion of the dye into the bar-chart channel when the pathogen is not detected.

Claims

1. A point-of-care testing (POCT) device for quantitative pathogen detection, comprising:

an inlet microwell for receiving a substance, the inlet microwell connected to a distribution channel to distribute the substance to an analyzing component;

the analyzing component comprising a first pathogen detection component, the first pathogen detection component comprising a platinum nanoparticle-labeled magnetic DNA-probe configured to recognize a first pathogen and then generate a fragmentary DNA-platinum nanoparticles, wherein the fragmentary DNA-platinum nanoparticles are configured to react with a substrate to generate gas to propel a dye through a bar-chart channel when a pathogen in the substance reacts with the platinum nanoparticle-labeled magnetic DNA-probe, wherein the platinum nanoparticle-labeled magnetic DNA-probe is configured to react with the first pathogen; and

at least one magnet configured to keep unreacted platinum nanoparticle-labeled magnetic DNA-probe in an amplification microwell, thereby inhibiting propulsion of the dye into the bar-chart channel when the pathogen is not detected.

2. The device of claim 1, wherein the distribution channel comprises a plurality of distribution channels and the analyzing component comprises a plurality of pathogen detection components, wherein each of the pathogen detection component is configured to detect a different pathogen, and wherein each of the pathogen detection components comprises the platinum nanoparticle-labeled magnetic DNA-probe configured to propel the dye through the bar-chart channel when the pathogen in the substance reacts with the platinum nanoparticle-labeled magnetic DNA-probe, wherein each of the platinum nanoparticle-labeled magnetic DNA-probes is configured to react with a different pathogen.

3. The device of claim 2, wherein each of the plurality of pathogen detection components is configured to receive a portion of a single sample received at the inlet microwell by rotating a spin unit (Status “

ON”

) to connect a common inlet to all bar-chart detection units.

4. The device of claim 2, wherein each of the plurality of pathogen detection components is isolated from the other ones of the plurality of pathogen detection components by rotating a spin unit (Status “

OFF”

) to disconnect a common inlet from the bar-chart channels such that a reaction to the pathogen in a first pathogen detection component does not propel the dye in a second pathogen detection component into the bar-chart channel corresponding to the second pathogen detection component.

5. The device of claim 2, wherein the at least one magnet comprises a magnet for each of a plurality of analyzing components.

6. The device of claim 1, wherein the platinum nanoparticle-labeled magnetic DNA-probe comprises a DNA hybridization between magnetic beads-complementary DNA and aptamer DNA-platinum nanoparticles.

7. The device of claim 1, further comprising:

a spin unit.

8. A multiplexed bar-chart spinchip for instrument free visual quantitative detection of multiple pathogens for point-of-care testing (POCT), comprising:

a layer-one sheet comprising a layer-one first surface and a layer-one second surface, the layer-one sheet further comprising a layer-one inlet connected to a plurality of layer-one branched channels and a plurality of layer-one exhaust outlets;

a layer-two sheet comprising a layer-two first surface and a layer-two second surface, the layer-two first surface in contact with the layer-one second surface, the layer-two sheet further comprising a plurality of layer-two sample inlets, a plurality of layer-two substrate inlets, a plurality of layer-two indicator inlets, a plurality of layer-two exhaust outlets, and a plurality of layer-two outlets;

a layer-three sheet comprising a layer-three first surface and a layer-three second surface, the layer-three first surface bonded to the layer-two second surface, a first surface of layer-three sheet further comprising a plurality of sample recognition microwells, a plurality of catalytic amplification microwells, a plurality of indicator microwells, a plurality of “

T”

-phase exchange channels, a plurality of connection channels, and a plurality of bar-chart channels, wherein each of the sample recognition microwells comprises a platinum nanoparticle-labeled magnetic DNA-probe, wherein the platinum nanoparticle-labeled magnetic DNA-probe in one of the sample recognition microwells is different from at least one of the other platinum nanoparticle-labeled magnetic DNA-probes in another one of the sample recognition microwells, wherein each of the sample recognition microwells is connected with a corresponding one of the catalytic amplification microwells and a corresponding one of the indicator microwells by a corresponding one of the “

T”

-phase exchange channels, wherein each of the connection channels is configured to specially connect the corresponding one of the sample recognition microwells and the corresponding one of the catalytic amplification microwells, wherein the plurality of sample recognition microwells, the plurality of catalytic amplification microwells, the plurality of indicator microwells, the plurality of “

T”

-phase exchange channels, the plurality of connection channels, and the plurality of bar-chart channels form a plurality of parallel microfluidic units;

a layer-four sheet comprising a layer-four first surface and a layer-four second surface, the layer-four first surface contacting the layer-three second surface, the layer-four second surface comprising a plurality of hollow microwells;

a plurality of magnets, each of the magnets residing in one of the hollow microwells; and

a layer-five sheet comprising a layer-five first surface and a layer-five second surface, the layer-five first surface contacting the layer-four second surface thereby securing the magnets in the hollow microwells;

wherein the magnets keep unreacted platinum nanoparticle-labeled magnetic DNA-probe in a corresponding one of the sample recognition microwells; and

wherein a dye is forced through one of the bar-chart channels when a sample is introduced with a pathogen corresponding to a particular platinum nanoparticle-labeled magnetic DNA-probe.

9. The multiplexed bar-chart spinchip of claim 8, wherein the platinum nanoparticle-labeled magnetic DNA-probes each comprise a DNA hybridization between magnetic beads-complementary DNA and aptamer DNA-platinum nanoparticles.

10. The multiplexed bar-chart spinchip of claim 8, wherein a distance that the dye moves is proportional to an analyte concentration, thus enabling quantitative detection of pathogens, without the aid of any detectors.

11. The multiplexed bar-chart spinchip of claim 8, wherein multiple different types of pathogens are quantitatively detected simultaneously from a single assay.

12. A point-of-care testing (POCT) device for quantitative pathogen detection, comprising:

an inlet microwell for receiving a substance;

an analyzing component configured to propel a dye through a bar-chart channel when a pathogen in the substance reacts with a catalyst-mediated magnetic DNA-probe; and

at least one magnet configured to keep unreacted catalyst-mediated magnetic DNA-probe in an amplification microwell, thereby inhibiting propulsion of the dye into the bar-chart channel when the pathogen is not detected;

wherein the inlet microwell is connected to a distribution channel to distribute the substance to the analyzing component.

13. The POCT device of claim 12, further comprising a catalyst nanoparticle-mediated magnetic DNA-probe, wherein the catalyst nanoparticle-mediated magnetic DNA-probe is configured to recognize the pathogen and generate fragmentary DNA-catalyst nanoparticles.

14. The POCT device of claim 13, wherein the fragmentary DNA-catalyst nanoparticles are configured to react with a substrate to generate gas to propel the dye through the bar-chart channel when the pathogen in the substance reacts with the catalyst nanoparticle-labeled magnetic DNA-probe.

15. The POCT device of claim 12, wherein the catalyst nanoparticle-labeled magnetic DNA-probe is configured to react with a first pathogen, and wherein the catalyst nanoparticle-labeled magnetic DNA-probe is configured to not react with a second pathogen.

16. The POCT device of claim 12, wherein the catalyst comprises platinum.

17. A method of fabricating a point-of-care testing (POCT) device for quantitative pathogen detection, the method comprising:

forming an inlet microwell for receiving a substance, the inlet microwell connected to a distribution channel to distribute the substance to an analyzing component;

forming the analyzing component comprising a first pathogen detection component, the first pathogen detection component comprising a platinum nanoparticle-labeled magnetic DNA-probe configured to recognize a first pathogen and then generate a fragmentary DNA-platinum nanoparticles, wherein the fragmentary DNA-platinum nanoparticles are configured to react with a substrate to generate gas to propel a dye through a bar-chart channel when a pathogen in the substance reacts with platinum nanoparticle-labeled magnetic DNA-probe, wherein the platinum nanoparticle-labeled magnetic DNA-probe is configured to react with the first pathogen; and

forming a layer comprising at least one magnet configured to keep unreacted platinum nanoparticle-labeled magnetic DNA-probe in an amplification microwell, thereby inhibiting propulsion of the dye into the bar-chart channel when the pathogen is not detected.