Minimum procedure system for the determination of analytes

• US 4,935,346 A
• Filed: 08/13/1986
• Issued: 06/19/1990
• Est. Priority Date: 08/13/1986
• Status: Expired due to Term

• Current Assignee: Lifescan Incorporated (Duv Holding Corporation)
• Original Assignee: Lifescan Incorporated (Duv Holding Corporation)
• Inventors: Phillips, Roger, Underwood, Ray, McGarraugh, Geoffery, Jurik, Frank
• Primary Examiner(s): Kepplinger, Esther M.
• Assistant Examiner(s): Spiegel, Carol A.
• Application Number:
  US06/896,418
• Time in Patent Office: 1,406 Days
• Field of Search: 435/4, 435/14, 435/25, 435/28, 435/805, 435/808, 435/810, 436/24, 436/46, 436/63, 436/95, 436/166, 436/169, 422/56, 422/57, 422/58, 422/61, 422/68
• US Class Current: 435/14
• CPC Class Codes: C12Q 1/54 involving glucose or galactose
• CPC Class Codes: G01N 2021/4757 Geometry 0/45° or 45/0°
• CPC Class Codes: G01N 21/314 with comparison of measurements at specific and non-specific wavelengths dual wavelength spectrometry G01J3/427
• CPC Class Codes: G01N 21/4738 Diffuse reflection precedence is given to G01N21/55 - G01N21/57 if specular component is taken into consideration, e.g. also for testing fluids, fibrous materials
• CPC Class Codes: G01N 21/8483 Investigating reagent band test-element handling not specific to a test method G01N33/4875; analytical elements specific to chemical analysis of biological material G01N33/52; autometer with reagent band G01N35/04
• CPC Class Codes: G01N 33/52 Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72
• CPC Class Codes: G01N 33/521 Single-layer analytical elements
• CPC Class Codes: G01N 33/523 the element being adapted for a specific analyte
• CPC Class Codes: G01N 33/66 involving blood sugars, e.g. galactose
• CPC Class Codes: G01N 35/1016 Control of the volume dispensed or introduced
• CPC Class Codes: Y02A 90/10 Information and communication technologies [ICT] supporting adaptation to climate change, e.g. for weather forecasting or climate simulation
• CPC Class Codes: Y10S 435/805 Test papers
• CPC Class Codes: Y10S 435/808 Optical sensing apparatus
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10T 436/112499 with sample on test slide
• CPC Class Codes: Y10T 436/144444 Glucose

Abstract

A method for determining the presence of an analyte in a fluid is described along with various components of an apparatus specifically designed to carry out the method. The method involves taking a reflectance reading from one surface of an inert porous matrix impregnated with a reagent that will interact with the analyte to produce a light-absorbing reaction product when the fluid being analyzed is applied to another surface and migrates through the matrix to the surface being read. Reflectance measurements are made at two separate wavelengths in order to eliminate interferences, and a timing circuit is triggered by an initial decrease in reflectance by the wetting of the surface whose reflectance is being measured by the fluid which passes through the inert matrix. The method and apparatus are particularly suitable for the measurement of glucose levels in blood without requiring separation of red blood cells from serum or plasma.

Claims

1. In a method for determining glucose in a blood sample employing a membrane and a signal-producing system which reacts with glucose to produce a light-absorptive dye product, said system being bound to the membrane, and in which the amount of said dye product is determined by means of a reflectance measurement from a surface of said membrane, an improvement which comprises:

applying an unmeasured whole blood sample to a first surface of a single-layer, substantially reflective, porous, hydrophilic membrane having pores of a size sufficient to exclude red blood cells and which contains said signal-producing system;

making said reflectance measurement on a second surface of said membrane other than the surface to which said sample is applied without removing excess sample or red blood cells from said first surface; and

determining the concentration of glucose in said sample from said reflectance measurement.

2. A method according to claim 1, wherein said signal-producing system produces a dye product which absorbs light at a wavelength different from a wavelength at which said red blood cells absorb and said reflectance measurement is made at two different wavelengths, one absorption wavelength reflectance measurement to correct for background absorbance due to red blood cells and the other reflectance measurement at the absorption wavelength of said dye product.

3. A method according to claim 2, wherein said two wavelengths are at about 635 and 700 nm.

4. A method according to claim 1, wherein said signal producing system comprises glucose oxidase, peroxidase, 3-methyl-2-benzothiazolinone hydrazone/3-(dimethylamino)benzoic acid in a weight ratio of about 1:

1 to about 4;

1 as an indicator, and a buffer providing a pH of 3.8 to 5.0.

5. The method of claim 4, wherein said buffer comprises 5-15 weight % citrate buffer dried in said membrane.

6. A method according to claim 1 wherein said membrane comprises polyamide and said signal producing system comprises glucose oxidase, peroxidase, and 3-methyl-2-benzothiazolinone hydrazone/3-(dimethylamino)benzoic acid.

7. The method of claim 1, wherein said pores have an average diameter of from about 0.1 to about 3.0 μ

m.

8. The method of claim 1, wherein said membrane consists essentially of polyamide and has a thickness of from about 0.01 to about 0.5 mm.

9. The method of claim 1, wherein said membrane consists essentially of polyamide and has a pore volume of less than 10 μ

l in said second surface.

10. A method for determining glucose comprising the sequential steps of:

(a) applying a whole blood sample to an application site on a reagent element wherein said reagent element comprises a single-layer, substantially reflective, porous, hydrophilic matrix which filters out red blood cells and to which is bound a signal-producing system comprising glucose oxidase, peroxidase and a dye indicator, which signal-producing system reacts with glucose to form a reaction dye product;

(b) allowing the sample to migrate to a reading site on said membrane different from said application site;

(c) monitoring reflectance at said reading site for a decrease in reflectance indicative of sample presence in said reading site in order to initiate timing of an incubation period;

and

(d) determining the change in reflectance at said reading site during the incubation period as a measure of dye product formed to determine the amount of glucose in said sample

wherein

all reflectance measurements at said reading site are performed without removing excess sample or red blood cells from said application site and at least one measurement is taken at a wavelength at which light is absorbed by said dye product.

11. A method according to claim 10, wherein said dye indicator is 3-methyl-2-benzothiazolinonehydrazone/3-(dimethylamino)benzoic acid.

12. A method according to claim 10, wherein said hydrophilic membrane comprises a polyamide.

13. A method according to claim 12, wherein said hydrophilic membrane is positively charged.

14. The method of claim 10, wherein said determination of the amount of glucose in said sample further includes determining the change in reflectance at a second wavelength to provide a correction for background absorbance at the dye product absorbing wavelength due to an interfering substance in said sample.

15. A method of determining analyte concentration in a liquid, which comprises:

quantitatively measuring baseline reflectance from a first surface of a reagent element comprising an inert, porous, hydrophilic, substantially reflective, single-layer matrix having pores of a size sufficient to exclude red blood cells and a reagent system which interacts with said analyte to produce a light-absorbing reaction product, said reagent system being impregnated in the pores of said matrix, prior to application of said liquid to said reagent element;

applying said liquid to a second surface of said reagent element and allowing said liquid to migrate from said second surface to said first surface;

quantitatively measuring reaction reflectance from said first surface of said reagent element without removing excess sample or non-migrating components of said sample from said second surface;

quantitatively measuring reflectance of interfering substances from said first surface of said reagent element using a wavelength of light reflected by interfering substances and different from the wavelength of light used to measure said reaction product reflectance in order to correct for background reflectance at the reaction product wavelength caused by interfering substances; and

calculating a value expressing said analyte concentration from said reflectance measurements.

16. The method of claim 15, wherein said matrix comprises a polyamide.

17. The method of claim 15, wherein the average diameter of the pores in said matrix is from 0.2 to 1.0 μ

m and said liquid is whole blood.

18. The method of claim 17, wherein said analyte is glucose, and said reagent produces a light-absorbing reaction product upon reacting with glucose.