Test device and method for quantitative measurement of an analyte in a liquid

• US 6,042,543 A
• Filed: 10/14/1997
• Issued: 03/28/2000
• Est. Priority Date: 03/11/1997
• Status: Expired due to Term

• Current Assignee: University of Minnesota
• Original Assignee: Regents of The University of Minnesota (University of Minnesota)
• Inventors: Hansen, Leland G., Warwick, Warren J.
• Primary Examiner(s): Hindenburg, Max
• Assistant Examiner(s): Natnithithadha, Navin
• Application Number:
  US08/950,523
• Time in Patent Office: 896 Days
• Field of Search: 600/362, 600/346, 600/348, 600/367, 600/309, 600/345, 600/347, 600/361, 600/365, 422/56, 422/58, 436/165
• US Class Current: 600/362
• CPC Class Codes: A61B 5/4266 sweat secretion
• CPC Class Codes: G01N 33/84 involving inorganic compounds or pH

Abstract

A device and method for determining the concentration of an analyte, particularly a chloride ion, in a liquid sample are particularly useful to screen for cystic fibrosis in an individual. In a preferred embodiment, the device takes the form of a patch comprising an absorbent material sandwiched between two liquid-impermeable layers. Chloride ions in the liquid sample complex with a chloride-detecting agent dispersed throughout the absorbent material to form a visible precipitate, preferably silver chloride. A pH indicator is also dispersed throughout the absorbent material, allowing the position of the water front to be monitored thereby permitting calculation of chloride concentration from a partially filled patch.

Claims

1. A device of substantially flat construction for application to the skin of a patient, collection of a test sample of sweat, and quantitative determination of the chloride concentration in the test sample of sweat, comprising:

a top transparent liquid-impermeable layer;

a bottom liquid-impermeable layer for contacting the skin comprising a centrally located inlet and a plurality of channels extending radially from the inlet for collecting the sweat and directing the sweat toward the inlet; and

a middle layer between the top and bottom layers comprising an absorbent material having substantially uniformly dispersed throughout at least a portion thereof (i) a liquid-detecting agent capable of detectably delimiting the liquid front formed as the liquid portion of the test sample migrates radially outwardly from the inlet through the substantially flat absorbent material, and (ii) a partitioning agent comprising a species capable of complexing with chloride ions present in the test sample to form a detectable precipitate disposed radially inwardly of said liquid front.

2. The device of claim 1 wherein the partitioning agent comprises silver chromate.

3. The device of claim 1 wherein the liquid-detecting substance comprises a pH indicator.

4. The device of claim 1 wherein the pH indicator comprises phenol red.

5. The device of claim 1 wherein the alkaline substance comprises sodium bicarbonate.

6. The device of claim 1 wherein the bodily fluid is sweat obtained from a patient suspected of being afflicted with cystic fibrosis.

16. The device of claim 1, wherein the inlet comprises an alkaline material capable of ensuring that the pH of the liquid sample entering the device is at least about pH 7.0.

27. A method for performing a diagnostic test for cystic fibrosis comprising:

inducing the production of sweat on the skin of a patient;

applying the bottom layer of the device of claim 1 to the skin of the patient such that the sweat enters the device through the inlet and diffuses through a portion of the absorbent material to yield a precipitation area comprising a detectable precipitate comprising the chloride ions, and a fill area detectably delimited by the liquid front;

obtaining a measurement of the precipitation area;

obtaining a measurement of the fill area; and

evaluating the precipitation area measurement and the fill area measurement to determine the concentration of chloride ions in the sweat, wherein the concentration of chloride ions in the sweat is used to diagnose the presence or absence of cystic fibrosis.

28. A diagnostic kit for screening for cystic fibrosis comprising at least one of the device of claim 1 and instructions for use of the device.

29. The diagnostic kit of claim 28 further comprising a known amount of a chloride salt, and at least one device of claim 1 to serve as a quality control.

30. The diagnostic kit of claim 28 further comprising an aqueous solution comprising a known concentration of a chloride salt, and at least one device of claim 1 to serve as a quality control.

31. The diagnostic kit of claim 30 wherein the chloride salt is sodium chloride.

32. The diagnostic kit of claim 28, further comprising a standard curve, a mathematical equation, or both, for use in determining chloride concentration in a test sample using a measurement of a test sample precipitation area, wherein the test sample precipitation area comprises a precipitate comprising chloride and the substance capable of visibly reacting with chloride.

7. A device for quantitatively determining the concentration of a chloride ion containing analyte in a liquid sample comprising:

a first liquid-impermeable layer;

a second liquid-impermeable layer comprising an inlet for the introduction of the liquid sample; and

a third layer between the first and second liquid-impermeable layers comprising an absorbent material comprising (i) a liquid-detecting agent capable of detectably delimiting the liquid front formed as the liquid migrates radially outwardly from the inlet through the absorbent material, and (ii) a partitioning agent capable of detectably removing the chloride ions in the analyte from the liquid sample through complexing with chloride ions present in the liquid of said sample.

8. The device of claim 7, wherein at least one of the first and second liquid impermeable layers is transparent.

9. The device of claim 7, wherein the partitioning agent is distributed substantially uniformly throughout at least a portion of the absorbent material.

10. The device of claim 7, wherein the partitioning agent comprises a species capable of forming a detectable complex comprising the analyte.

11. The device of claim 10, wherein the detectable complex comprises a precipitate comprising the analyte.

12. The device of claim 7, wherein the partitioning agent is bound to the absorbent material.

13. The device of claim 12, wherein the bound partitioning agent comprises a species capable of binding the analyte to form a detectable complex bound to the absorbent material.

14. The device of claim 7, wherein the analyte is a chloride ion.

15. The device of claim 14, wherein the partitioning agent comprises silver chromate.

17. A device for quantitatively determining the concentration of chloride ion present in an analyte in a liquid sample comprising:

a first liquid-impermeable layer;

a second liquid-impermeable layer comprising an inlet for the introduction of the liquid sample; and

a third layer between the first and second liquid-impermeable layers comprising an absorbent material comprising (i) a measurable fill area delimited by a detectable water front formed by a liquid sample introduced into the device through the inlet for radial outward migration therefrom; and

(ii) a measurable detection area comprising a component capable of species specific complexing with chloride ions for forming an immobilized or insoluble detectable complex comprising the analyte.

21. The method of claim 17, further comprising (c) inserting into the inlet a paste or plug comprising an alkaline substance capable of ensuring that the pH of a liquid sample entering the device is at least about pH 7.0.

18. A method for preparing a device for quantitatively determining the concentration of chloride ions present in an analyte in a liquid sample comprising:

(a) distributing throughout at least a portion of an absorbent material each of;

(i) a liquid-detecting agent, and

(ii) a partitioning agent capable of detectably removing chloride present in the analyte from the liquid sample; and

(b) positioning the absorbent material between a first liquid-impermeable layer and a second liquid-impermeable layer comprising an inlet for introduction of the liquid sample, wherein the liquid-detecting substance is capable of detectably delimiting the liquid front formed as the liquid containing chloride ion migrates from the inlet through the absorbent material.

19. The method of claim 18, wherein the partitioning agent comprises silver chromate.

20. The method of claim 19, wherein the silver chromate is formed in situ in the absorbent material.

22. A method for determining the concentration of an analyte in a liquid sample comprising:

providing a device comprising a first liquid-impermeable layer, a second liquid-impermeable layer comprising an inlet, and a third layer between the first and second liquid-impermeable layers, said third layer comprising an absorbent material comprising each of a liquid-detecting agent and a partitioning agent;

introducing an amount of the liquid sample into the inlet of the device such that the liquid diffuses through the absorbent material so as to contact both the liquid-detecting agent and the partitioning agent, wherein contact of the liquid with the liquid-detecting agent allows detection of a water front delimiting a fill area of the absorbent material, and wherein contact of the liquid with the partitioning agent causes formation of a detectable complex comprising the analyte thereby effecting removal of the analyte from the liquid, the area of formation of said detectable complex constituting a detection area of the absorbent material;

obtaining a measurement of the fill area;

obtaining a measurement of the detection area; and

evaluating the fill area measurement and the detection area measurement to determine the concentration of the analyte in the liquid sample.

23. The method of claim 22, wherein the detectable complex comprises a precipitate comprising the analyte, and wherein the detection area is a precipitation area.

24. The method of claim 22, wherein the evaluation step comprises evaluating the fill area measurement and the detection area measurement in view of an empirically derived standard curve to determine the concentration of the analyte in the liquid sample.

25. The method of claim 22, wherein a computer is used to obtain the detection area measurement and the fill area measurement.

26. The method of claim 22, wherein a computer is used to evaluate the detection area measurement and the fill area measurement.

33. A device of substantially flat construction for application to the skin of a patient, collection of a test sample of sweat, and determination of the chloride concentration in the test sample of sweat, comprising:

a top transparent liquid-impermeable layer;

a bottom liquid-impermeable layer for contacting the skin comprising a centrally located inlet and a plurality of channels extending radially from the inlet for collecting the sweat and directing the sweat toward the inlet;

a middle layer between the top and bottom layers comprising an absorbent material having substantially uniformly dispersed throughout at least a portion thereof (i) a liquid-detecting agent capable of detectably delimiting the liquid front formed as the test sample migrates from the inlet through the substantially flat absorbent material, and (ii) a partitioning agent comprising a species capable of complexing with chloride ions to form a detectable precipitate; and

wherein said inlet comprises an alkaline substance capable of ensuring that the pH of the sweat entering the device is at least about pH 7.0.

34. A device for determining the concentration of an analyte in a liquid sample comprising:

a first liquid-impermeable layer;

a second liquid-impermeable layer comprising an inlet for the introduction of the liquid sample;

a third layer between the first and second liquid-impermeable layers comprising an absorbent material comprising (i) a liquid-detecting agent capable of detectably delimiting the liquid front formed as the liquid migrates from the inlet through the absorbent material, and (ii) a partitioning agent capable of detectably removing the analyte from the liquid sample; and

the liquid-detecting agent comprises a pH indicator.

35. A method for preparing a device for determining the concentration of an analyte in a liquid sample comprising:

(a) distributing throughout at least a portion of an absorbent material each of

(i) a liquid-detecting agent, and

(ii) a partitioning agent capable of detectably removing the analyte from the liquid sample;

(b) positioning the absorbent material between a first liquid-impermeable layer and a second liquid-impermeable layer comprising an inlet for introduction of the liquid sample, wherein the liquid-detecting substance is capable of detectably delimiting the liquid front formed as the liquid migrates from the inlet through the absorbent material; and

(c) the liquid-detecting substance comprises a pH indicator.