Microfluidics based assay device

• US 9,341,620 B2
• Filed: 02/07/2012
• Issued: 05/17/2016
• Est. Priority Date: 02/07/2011
• Status: Active Grant

• Current Assignee: Lumiradx UK Ltd
• Original Assignee: Multi-Sense Technologies Limited (LumiraDx Ltd. (Cayman Islands))
• Inventors: Lowe, Phillip, McGuigan, Brian, Keatch, Steven Alexander
• Primary Examiner(s): BROWN, MELANIE YU
• Application Number:
  US13/983,650
• Publication Number:
  US 20140017709A1
• Time in Patent Office: 1,561 Days
• US Class Current: 1/1
• CPC Class Codes: B01L 2200/04 Exchange or ejection of cartridges, containers or reservoirs
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/161 Control and use of surface tension forces, e.g. hydrophobic, hydrophilic
• CPC Class Codes: B01L 2400/043 magnetic forces
• CPC Class Codes: B01L 2400/0688 surface tension valves, capillary stop, capillary break
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• 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 1/001 Enzyme electrodes
• CPC Class Codes: C12Q 1/006 for glucose
• CPC Class Codes: C12Y 101/03004 Glucose oxidase (1.1.3.4)
• CPC Class Codes: G01N 2333/904 acting on CHOH groups as donors, e.g. glucose oxidase, lactate dehydrogenase (1.1)
• CPC Class Codes: G01N 27/3277 being a redox reaction, e.g. detection by cyclic voltammetry voltammetry per se G01N27/42, G01N27/48
• CPC Class Codes: G01N 33/54326 Magnetic particles
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/5438 Electrodes

Abstract

A subtractive corrective assay device and methodology, whereby ail required binding and label detection reagents are initially located within the detection zone. Application of a magnetic field is used to selectively remove bound label from the detection zone by means of paramagnetic particles. The relationship between measured label concentration before and after the application of a magnetic field within the detection zone is used to accurately measure analyte concentration within the sample.

Claims

1. A method of conducting an assay on a sample, the method comprising the steps of:

introducing a fluid sample into a microfluidic cartridge such that any analyte present in the sample is capable of being bound by an analyte binding agent and a label present in the cartridge, the cartridge comprising;

a sample port for introducing said sample into the cartridge,

a substrate comprising one or more microfluidic channels disposed therein, a binding agent within said channel(s) for binding any of said analyte within the sample, and a label for use in detecting an amount of the analyte present in the sample; and

a detection area within or wholly comprising an area in which analyte, analyte binding agent, and label binding occurs to produce analyte binding agent/analyte/label complexes ,

detecting a total level of the label that is complexed or uncomplexed to the analyte present in the detection area in order to obtain a first reference/control value;

removing any analyte binding agent/analyte/label complexes from the detection area;

detecting a level of any uncomplexed label and any unreacted label that remains in the detection area after removal of said analyte binding agent/analyte/label complexes; and

determining said analyte concentration by subtracting the level of uncomplexed/unreacted label from the first reference/control value.

2. The method according to claim 1, wherein introduction of the sample to the cartridge causes at least a portion of the binding agent and the label to be brought into solution or suspension within the detection area.

3. The method according to claim 2, wherein the detection is carried out electrochemically.

4. The method according to claim 3, wherein an oxidising potential or a reduction potential is used to measure a level of the label.

5. The method according to claim 4,

wherein the oxidising potential or the reduction potential is applied to reduce any background effects, such as to eliminate or minimise effect of electrochemical interferents which may be present in the sample,

wherein the effect of the electrochemical interferents may include generating a background signal or minimising a current generated by an enzyme during the binding incubation phase.

6. The method according to claim 3, wherein glucose oxidase is employed as the label.

7. The method according to claim 6, further comprising the step of:

using a peroxidise and a mediator wherein the mediator is capable of conversion between an oxidised form and a reduced form and is capable of electrochemical detection.

8. The method according to claim 1, wherein the binding agent is coupled or otherwise bound to a particle which is capable of being complexed with the analyte present in the sample and which can be removed, when complexed or uncomplexed with the analyte, from the detection area, by suitable means.

9. The method according to claim 8, wherein the particle is a paramagnetic particle and the suitable means is a magnet or an electromagnet.

10. The method according to claim 1, wherein the sample is a sample of whole blood.

11. The method according to claim 1, wherein detection is carried out directly on the sample, without any separation, washing step, or dilution of the sample by a further fluid being carried out.

12. The method according to claim 1, wherein a decrease in a signal before and after removal of the bound analyte/label from the detection area is detected, and wherein the signal is proportional to the amount of the analyte present in the sample.

13. The method according to claim 7, wherein the peroxidase is HRP.

14. The method according to claim 7, wherein the mediator is ferrocyanide or ABTS.