In vitro diagnostic assay methods

Patent metadata

Field	Value
Publication number	US-2022011314-A1
Application number	US-201917273641-A
Country	US
Kind code	A1
Filing date	Sep 5, 2019
Priority date	Sep 6, 2018
Publication date	Jan 13, 2022
Grant date	—

Abstract

An in vitro diagnostic assay method, comprising: providing a sample which may comprise target analyte(s); contacting the sample with a fluorescent label whose fluorescence can be externally modulated, such that the fluorescent label is associated with target analyte(s), if present, to form a fluorescent label-analyte complex; and detecting the fluorescent label-analyte complex; and device, kit and solid phase for performing an in vitro diagnostic assay method.

First claim

1 . An in vitro diagnostic assay method, comprising:
providing a sample which may comprise target analyte(s); contacting the sample with a fluorescent label whose fluorescence can be externally modulated, such that the fluorescent label is associated with target analyte(s), if present, to form a fluorescent label-analyte complex; and detecting the fluorescent label-analyte complex.

2 . Assay method according to claim 1 , further comprising:
a) exciting the fluorescent label using a suitable light source, and detecting the resulting fluorescence; b) modulating the fluorescence of the fluorescent label and detecting the resulting fluorescence to obtain a fluorescence time series comprising the fluorescence results from steps a) and b); and c) optionally repeating the modulation step one or more times and detecting the resulting fluorescence to obtain a fluorescence time series comprising the fluorescence results from steps a), b) and c).

3 . Assay method according to claim 2 , which comprises applying a frequency domain analysis to the fluorescence time series acquired in step b) or c), optionally wherein the frequency domain analysis comprises applying a lock-in amplification algorithm or Fourier transform to the fluorescence time series.

4 . (canceled)

5 . Assay method according to claim 1 , wherein the fluorescent label comprises a material having a nitrogen vacancy centre, wherein said material having a nitrogen vacancy centre comprises fluorescent nanodiamonds (FNDs).

6 . Assay method according to claim 5 , wherein the FNDs comprise a hydrophilic layer comprising a hydrophilic polymer, optionally wherein the hydrophilic polymer comprises polyglycerol (PG) or polyethylene glycol (PEG).

7 . (canceled)

8 . Assay method according to claim 5 , wherein the FNDs comprise probe(s) suitable for associating with the target analyte(s), optionally wherein said probe(s) comprise a peptide, protein, antibody, nucleic acid, oligonucleotide, aptamer, DNA, RNA, carbohydrate, small molecule (for example hormone, steroid, drug, small molecule less than 100 kDa) or cell.

9 . (canceled)

10 . Assay method according to claim 1 , wherein the assay method comprises a microfluidic flow assay, ELISA, qPCR, microarray, isothermal amplification, dot-blot, flow cytometry or fluorescence-activated cell sorting (FACS).

11 . Assay method according to claim 1 , wherein the sample is applied to a solid phase comprising a multiwell plate, beads, column, photonic device, microfluidic chip or capillary bed for microfluidic flow assay.

12 . Assay method according to claim 1 , which is a lateral flow assay or vertical flow assay.

13 . Assay method according to claim 2 , wherein the fluorescence of the fluorescent label is modulated by a microwave field or a magnetic field, optionally wherein the fluorescent label comprises a nitrogen vacancy centre, and a microwave resonator with resonant frequency of 1-40 GHz is used to enhance the coupling between the label and the applied modulated microwaves.

14 . (canceled)

15 . A device for performing an in vitro diagnostic assay, wherein said device comprises:
a solid phase according to claim 23 ; an excitation light source; a fluorescence modulator; and a fluorescence detector.

16 . Device according to claim 15 , further comprising a controller/processing unit, preferably configured to apply frequency-domain analysis to the results obtained from fluorescence detection.

17 . Device according to claim 15 , wherein:
the fluorescence detector comprises a portable reader; and/or the fluorescence modulator comprises a microwave signal generator; and optionally, a microwave resonator with resonant frequency of 1-40 GHz.

18 . (canceled)

19 . Device according to claim 15 , wherein:
the fluorescent label comprises a material having a nitrogen vacancy centre, wherein said material having a nitrogen vacancy centre comprises fluorescent nanodiamonds (FNDs) as defined in any one of claims 5 to 9 ; and/or wherein the solid phase comprises a multiwell plate, beads, column, photonic device, microfluidic chip or capillary bed for microfluidic flow assay.

20 . Device according to claim 15 , comprising a lateral flow assay or vertical flow assay.

21 . A kit for performing an in vitro diagnostic assay, wherein said kit comprises:
a solid phase comprising a fluorescent label whose fluorescence can be externally modulated, or a solid phase and a separate liquid suspension or solution comprising a fluorescent label whose fluorescence can be externally modulated.

22 . A kit according to claim 21 , further comprising:
an excitation light source; a fluorescence modulator; and a fluorescence detector.

23 . A solid phase comprising a fluorescent label whose fluorescence can be externally modulated, wherein said solid phase is selected from a multiwell plate, beads, column, photonic device, microfluidic chip or capillary bed for microfluidic flow assay.

24 . Solid phase according to claim 23 , which is a lateral flow assay or vertical flow assay.

25 . Solid phase according to claim 23 , wherein the fluorescent label comprises a material having a nitrogen vacancy centre, wherein said material having a nitrogen vacancy centre comprises fluorescent nanodiamonds (FNDs).

Assignees

• Ucl Business Ltd

Inventors

• Miller Ben
• Bezinge Leonard
• Dobson Peter
• Dold Gavin
• Morton John
• Mckendry Rachel

Classifications

• G01N21/6428 Primary

  Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53) · CPC title

• G01N33/587

  Nanoparticles · CPC title

• G01N33/582 Primary

  with fluorescent label · CPC title

• G01N2021/6439

  with indicators, stains, dyes, tags, labels, marks · CPC title