Personal glucose meters for detection and quantification of a broad range of analytes

Patent metadata

Field	Value
Publication number	US-8945943-B2
Application number	US-201113699578-A
Country	US
Kind code	B2
Filing date	May 26, 2011
Priority date	May 26, 2010
Publication date	Feb 3, 2015
Grant date	Feb 3, 2015

Abstract

A general methodology for the development of highly sensitive and selective sensors that can achieve portable, low-cost and quantitative detection of a broad range of targets using only a personal glucose meter (PGM) is disclosed. The method uses recognition molecules that are specific for a target agent, enzymes that can convert an enzyme substrate into glucose, and PGM. Also provided are sensors, which can include a solid support to which is attached a recognition molecule that permits detection of a target agent, wherein the recognition molecule specifically binds to the target agent in the presence of the target agent but not significantly to other agents as well as an enzyme that can catalyze the conversion of a substance into glucose, wherein the enzyme is attached directly or indirectly to the recognition molecule, and wherein in the presence of the target agent the enzyme can convert the substance into glucose. The disclosed sensors can be part of a lateral flow device. Methods of using such sensors for detecting target agents are also provided.

First claim

We claim:

1. A lateral flow device, comprising:
a first solid support to which is attached a first nucleic acid molecule that specifically binds to a target agent but not significantly to other agents;
an enzyme attached to second nucleic acid molecule, wherein the second nucleic acid molecule is complementary to a portion of the first nucleic acid molecule, thereby generating an enzyme conjugate, wherein the enzyme conjugate is hybridized to the first nucleic acid molecule, wherein the enzyme catalyzes the conversion of a substance into glucose, and wherein in the presence of the target agent the enzyme conjugate is released and separated from the first nucleic acid molecule, and the enzyme converts the substance into glucose; and
the substance that is converted into glucose by the enzyme, wherein the substance is soluble and attached to a second solid support.

2. The lateral flow device of claim 1 , wherein the first solid support comprises a bead and a conjugation pad, wherein the first nucleic acid molecule is attached directly to the bead, and wherein the bead is adsorbed to the conjugation pad.

3. The lateral flow device of claim 1 , wherein the first and/or the second solid support comprises nitrocellulose.

4. The lateral flow device of claim 1 , wherein the target agent comprises a metal ion, microbe, cytokine, hormone, cell, nucleic acid molecule, spore, protein, recreational drug, or toxin.

5. The lateral flow device of claim 1 , wherein:
the enzyme is an invertase, sucrase, or sucrase-isomaltase and the substance is sucrose,
the enzyme is a maltase and the substance is maltose,
the enzyme is a trehalase and the substance is trehalose,
the enzyme is an amylase and the substance is starch, or
the enzyme is a cellulase and the substance is cellulose.

6. The lateral flow device of claim 1 , wherein:
the first solid support comprises a conjugation pad;
the second solid support comprises a membrane; and
the lateral flow device further comprises:
a sample or wicking pad; and
an absorption pad.

7. The lateral flow device of claim 6 , wherein the enzyme is invertase and the substance that is converted into glucose is sucrose.

8. The lateral flow device of claim 1 , wherein the first nucleic acid is an enzyme strand and the second nucleic acid is a substrate strand of a DNAzyme or aptazyme.

9. The lateral flow device of claim 4 , wherein the metal ion is a heavy metal.

10. The lateral flow device of claim 9 , wherein the heavy metal is mercury (Hg 2+ ), cadmium (Cd 2+ ), arsenic (As 2+/3+ ), chromium (Cr 5+/6+ ), thallium (Tl +/3+ ), uranium (UO 2 2+ ), plutonium (Pu 3+/4+ , PuO 2+ , PuO 2 2+ , PuO 5 2+ ), or lead (Pb 2+ ).

11. The lateral flow device of claim 4 , wherein the metal ion is a nutritional metal.

12. The lateral flow device of claim 11 , wherein the nutritional metal is calcium (Ca 2+ ), iron (Fe 3+/4+ ), cobalt (Co 2+ ), magnesium (Mg 2+ ), manganese (Mn 2+ ), molybdenum (MoO 4 2+ ), zinc (Zn 2+ ), cadmium (Cd 2+ ), or copper (Cu +/2+ ).

13. A kit comprising:
one or more lateral flow devices of claim 1 ; and
one or more of a buffer or a chart for correlating detected glucose level and amount of target agent present.

14. A method for detecting a target agent, comprising:
contacting one or more lateral flow devices of claim 1 with a sample under conditions sufficient to allow the target agent in the sample to flow through the lateral flow device and bind to the first nucleic acid molecule present on the first solid support;
forming a target agent-nucleic acid complex, wherein formation of the target agent-nucleic acid complex results in the release of the enzyme conjugate from the first nucleic acid molecule;
allowing the enzyme of the enzyme conjugate to interact with the substance that is converted into glucose, thereby generating glucose; and
detecting glucose, wherein detection of glucose indicates the presence of the target agent in the sample, and an absence of detected glucose indicates the absence of the target agent in the sample.

15. The method of claim 14 , further comprising quantifying the target agent, wherein a level of glucose detected indicates an amount of target agent present.

16. The method of claim 14 , wherein
the enzyme comprises invertase, sucrase, or sucrase-isomaltase and the substance that the enzyme converts into glucose comprises sucrose, or
the enzyme comprises maltase and the substance that the enzyme converts into glucose comprises maltose, or
the enzyme comprises trehalase and the substance that the enzyme converts into glucose comprises trehalose, or
the enzyme comprises cellulase and the substance that the enzyme converts into glucose comprises cellulose, or
the enzyme comprises amylase and the substance that the enzyme converts into glucose comprises starch.

17. The method of claim 14 , wherein the glucose is detected using a personal glucose meter.

18. A lateral flow device, comprising:
a first region of a solid support to which is attached a first nucleic acid molecule that specifically binds to a target agent but not significantly to other agents;
an enzyme attached to a second nucleic acid molecule, wherein the second nucleic acid molecule is complementary to a portion of the first nucleic acid molecule, thereby generating an enzyme-conjugate, wherein the enzyme-conjugate is hybridized to the first nucleic acid molecule, wherein the enzyme catalyzes the conversion of a substance into glucose, and wherein in the presence of the target agent the enzyme-conjugate is released and separated from the first nucleic acid molecule, and the enzyme converts the substance into glucose; and
the substance that is converted into glucose by the enzyme, wherein the substance is soluble and attached to a second region of the solid support.

19. The lateral flow device of claim 18 , wherein the first nucleic acid is an enzyme strand and the second nucleic acid is a substrate strand of a DNAzyme or aptazyme.

20. The lateral flow device of claim 18 , wherein the target agent comprises a metal ion, microbe, cytokine, hormone, cell, nucleic acid molecule, spore, protein, recreational drug, or toxin.

21. The lateral flow device of claim 18 , wherein the solid support comprises a membrane, and wherein the nucleic acid molecule is attached directly to the membrane.

22. The lateral flow device of claim 18 , wherein:
the enzyme is an invertase, sucrase, or sucrase-isomaltase and the substance is sucrose,
the enzyme is a maltase and the substance is maltose,
the enzyme is a trehalase and the substance is trehalose,
the enzyme is an amylase and the substance is starch, or
the enzyme is a cellulase and the substance is cellulose.

23. A method for detecting a target agent, comprising:
contacting one or more lateral flow devices of claim 18 with a sample under conditions sufficient to allow the target agent in the sample to flow through the lateral flow device and bind to the first nucleic acid molecule present on the solid support;
forming a target agent-nucleic acid complex, wherein formation of the target agent-nucleic acid complex results in the release of the enzyme-conjugate from the first nucleic acid molecule;
allowing the enzyme of the enzyme-conjugate to interact with the substance that is converted into glucose, thereby generating glucose; and
detecting glucose, wherein detection of glucose indicates the presence of the target agent in the sample, and an absence of

Assignees

• Lu Yi
• Xiang Yu
• Univ Illinois

Inventors

• Lu Yi
• Xiang Yu

Classifications

• C12Q1/54 Primary

  involving glucose or galactose · CPC title

• C12Q1/34

  involving hydrolase · CPC title

• G01N33/558 Primary

  using diffusion or migration of antigen or antibody {(immunochromatographic test strips G01N33/54387)} · CPC title