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USRE42924E1 - Electrochemical biosensor test strip - Google Patents

️Tue Nov 15 2011

USRE42924E1 - Electrochemical biosensor test strip - Google Patents

Electrochemical biosensor test strip Download PDF

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Publication number

USRE42924E1

USRE42924E1 US10/692,031 US69203103A USRE42924E US RE42924 E1 USRE42924 E1 US RE42924E1 US 69203103 A US69203103 A US 69203103A US RE42924 E USRE42924 E US RE42924E Authority

United States

Prior art keywords

test

test strip

insulating substrate

capillary channel

roof

Prior art date

1997-12-05

Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)

Expired - Lifetime

Application number

US10/692,031

Inventor

William F. Crismore

Nigel A. Surridge

Daniel R. McMinn

Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)

Roche Diabetes Care Inc

Original Assignee

Roche Operations Ltd

Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)

1997-12-05

Filing date

2003-10-23

Publication date

2011-11-15

2003-10-23 Application filed by Roche Operations Ltd filed Critical Roche Operations Ltd

2003-10-23 Priority to US10/692,031 priority Critical patent/USRE42924E1/en

2004-08-16 Assigned to ROCHE DIAGNOSTICS OPERATIONS, INC. reassignment ROCHE DIAGNOSTICS OPERATIONS, INC. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: ROCHE DIAGNOSTICS CORPORATION

2007-06-13 Assigned to CORANGE INTERNATIONAL LIMITED reassignment CORANGE INTERNATIONAL LIMITED ASSIGNOR HEREBY CONFIRMS THE ASSIGNMENT TO CORANGE INTERNATIONAL LIMITED OF AN UNDIVIDED ONE-HALF INTEREST IN EACH OF THE APPLICATIONS Assignors: ROCHE DIAGNOSTICS OPERATIONS, INC.

2007-12-21 Assigned to BOEHRINGER MANNHEIM CORPORATION reassignment BOEHRINGER MANNHEIM CORPORATION ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: EARL, ROBERT KITCHEL, HEALD, BRIAN A., SURRIDGE, NIGEL A., DELK, R. DALE, DIEBOLD, ERIC R., BURKE, DAVID W., BODENSTEINER, RICHARD J., CRISMORE, WILLIAM F., MCMINN, DANIEL R., HO, JIAXIONG JASON

2007-12-21 Assigned to ROCHE DIAGNOSTICS OPERATIONS, INC. reassignment ROCHE DIAGNOSTICS OPERATIONS, INC. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: ROCHE DIAGNOSTICS CORPORATION

2007-12-21 Assigned to ROCHE DIAGNOSTICS CORPORATION reassignment ROCHE DIAGNOSTICS CORPORATION MERGER (SEE DOCUMENT FOR DETAILS). Assignors: BOEHRINGER MANNHEIM CORPORATION, ROCHE DIAGNOSTIC SYSTEMS, INC.

2009-05-05 Assigned to ROCHE OPERATIONS LTD. reassignment ROCHE OPERATIONS LTD. CHANGE OF NAME (SEE DOCUMENT FOR DETAILS). Assignors: CORANGE INTERNATIONAL LIMITED

2011-11-15 Publication of USRE42924E1 publication Critical patent/USRE42924E1/en

2011-11-15 Application granted granted Critical

2015-06-23 Assigned to ROCHE DIABETES CARE, INC. reassignment ROCHE DIABETES CARE, INC. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: ROCHE DIAGNOSTICS OPERATIONS, INC.

2017-12-05 Anticipated expiration legal-status Critical

Status Expired - Lifetime legal-status Critical Current

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Classifications

- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/001—Enzyme electrodes
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3271—Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood
- G01N27/3272—Test elements therefor, i.e. disposable laminated substrates with electrodes, reagent and channels
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
- G01N33/5438—Electrodes
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S435/00—Chemistry: molecular biology and microbiology
- Y10S435/817—Enzyme or microbe electrode

Definitions

This invention relates to a biosensor and its use in the detection or measurement of analytes in fluids.
test strips including electrochemical biosensor test strips, for measuring the amount of an analyte in a fluid.
test strips have been used by diabetics and health care professionals for monitoring their blood glucose levels.
the test strips are usually used in conjunction with a meter, which measures light reflectance, if the strip is designed for photometric detection of a dye, or which measures some electrical property, such as electrical current, if the strip is designed for detection of an electroactive compound.
test strip When the test strip is a capillary fill device, that is, when the chemical reaction chamber of the test strip is a capillary space, particular problems can occur with filling the chamber smoothly and sufficiently with the liquid sample to be tested. Due to the smallness of the capillary space and the composition of materials used to make the test strip, the test sample may hesitate entering the capillary reaction chamber. Further, insufficient sample may also be drawn into the capillary reaction chamber, thereby resulting in an inaccurate test result. It would be very useful if such problems could be minimized.
the electrochemical, biosensor test strip of the present invention provides solutions to these above-stated problems found in prior art test strips.
the first new feature is an indentation along one edge of the test strip for easy identification of the sample application port for vision impaired persons or for use in zero or low lighting conditions.
the test strip has a capillary test chamber, and the roof of the test chamber includes the second new feature of the biosensor test strip.
the second new feature is a transparent or translucent window which operates as a “fill to here” line, thereby identifying when enough test sample (a liquid sample, such as blood) has been added to the test chamber to accurately perform a test.
the window defines the minimum sample amount, or dose, required to accurately perform a test, and, therefore, represents a visual failsafe which reduces the chances of erroneous test results due to underdosing of a test strip.
FIG. 1 is an exploded view of a preferred embodiment of the present invention.
FIG. 4 is a cross sectional view of the test strip of FIG. 2 through line 28 — 28 .
calibration curve 30 ( FIG. 6 ) may be constructed. This calibration will be stored in the Read Only Memory (ROM) key of the meter and will be applicable to a particular lot of test strips. Lines 31 and 32 in FIG. 6 represent other hypothetical calibration curves for two other different lots of test strips. Calibration for these biosensor lots would generate slightly different values for C and d in the above algorithm.
ROM Read Only Memory

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Health & Medical Sciences (AREA)
Life Sciences & Earth Sciences (AREA)
Chemical & Material Sciences (AREA)
Immunology (AREA)
Engineering & Computer Science (AREA)
Molecular Biology (AREA)
General Health & Medical Sciences (AREA)
Organic Chemistry (AREA)
Physics & Mathematics (AREA)
Biochemistry (AREA)
Hematology (AREA)
Analytical Chemistry (AREA)
Wood Science & Technology (AREA)
Microbiology (AREA)
Zoology (AREA)
Biophysics (AREA)
Pathology (AREA)
General Physics & Mathematics (AREA)
Biotechnology (AREA)
Urology & Nephrology (AREA)
Biomedical Technology (AREA)
Proteomics, Peptides & Aminoacids (AREA)
Cell Biology (AREA)
Electrochemistry (AREA)
Food Science & Technology (AREA)
Medicinal Chemistry (AREA)
General Engineering & Computer Science (AREA)
Bioinformatics & Cheminformatics (AREA)
Chemical Kinetics & Catalysis (AREA)
Genetics & Genomics (AREA)
Investigating Or Analysing Biological Materials (AREA)
Investigating Or Analyzing Materials By The Use Of Electric Means (AREA)
Apparatus Associated With Microorganisms And Enzymes (AREA)
Treatments Of Macromolecular Shaped Articles (AREA)
Polishing Bodies And Polishing Tools (AREA)
Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Measurement Of The Respiration, Hearing Ability, Form, And Blood Characteristics Of Living Organisms (AREA)

Abstract

An electrochemical biosensor test strip with four new features. The test strip includes an indentation for tactile feel as to the location of the strips sample application port. The sample application port leads to a capillary test chamber, which includes a test reagent. The wet reagent includes from about 0.2% by weight to about 2% by weight polyethylene oxide from about 100 kilodaltons to about 900 kilodaltons mean molecular weight, which makes the dried reagent more hydrophilic and sturdier to strip processing steps, such as mechanical punching, and to mechanical manipulation by the test strip user. The roof of the capillary test chamber includes a transparent or translucent window which operates as a “fill to here” line, thereby identifying when enough test sample (a liquid sample, such as blood) has been added to the test chamber to accurately perform a test. The test strip may further include a notch located at the sample application port. The notch reduces a phenomenon called “dose hesitation”.

Description

Notice: More than one reissue application has been filed for the reissue of U.S. Pat. No. 5,997,817. The reissue applications are application Ser. Nos. 10/692,031 (the present application), 10/008,788, 10/409,721 and 10/693,305, all of which are divisional reissues of U.S. Pat. No. 5,997,817.

FIELD OF THE INVENTION

This invention relates to a biosensor and its use in the detection or measurement of analytes in fluids.

BACKGROUND OF THE INVENTION

The prior art includes test strips, including electrochemical biosensor test strips, for measuring the amount of an analyte in a fluid.

Particular use of such test strips has been made for measuring glucose in human blood. Such test strips have been used by diabetics and health care professionals for monitoring their blood glucose levels. The test strips are usually used in conjunction with a meter, which measures light reflectance, if the strip is designed for photometric detection of a dye, or which measures some electrical property, such as electrical current, if the strip is designed for detection of an electroactive compound.

However, test strips that have been previously made present certain problems for individuals who use them. For example, test strips are relatively small and a vision impaired diabetic may have great difficulty properly adding a sample of blood to the sample application area of the test strip. It would be useful for the test strip to be made so that vision impaired persons could easily dose the test strip.

When the test strip is a capillary fill device, that is, when the chemical reaction chamber of the test strip is a capillary space, particular problems can occur with filling the chamber smoothly and sufficiently with the liquid sample to be tested. Due to the smallness of the capillary space and the composition of materials used to make the test strip, the test sample may hesitate entering the capillary reaction chamber. Further, insufficient sample may also be drawn into the capillary reaction chamber, thereby resulting in an inaccurate test result. It would be very useful if such problems could be minimized.

Finally, test strips, especially those used by diabetics for measuring blood glucose are mass produced. Processes, such as mechanical punching, used to make these test strips can cause a test reagent that has been dried onto a surface of the testing area to crack or break, thereby causing reagent loss or improper placement of the reagent within the strip. It would also be useful to design a test reagent that could withstand processing steps, such as mechanical punching.

The electrochemical, biosensor test strip of the present invention provides solutions to these above-stated problems found in prior art test strips.

SUMMARY OF THE INVENTION

The invention is an improved electrochemical biosensor test strip with four new, highly advantageous features.

The first new feature is an indentation along one edge of the test strip for easy identification of the sample application port for vision impaired persons or for use in zero or low lighting conditions.

The test strip has a capillary test chamber, and the roof of the test chamber includes the second new feature of the biosensor test strip. The second new feature is a transparent or translucent window which operates as a “fill to here” line, thereby identifying when enough test sample (a liquid sample, such as blood) has been added to the test chamber to accurately perform a test. The window defines the minimum sample amount, or dose, required to accurately perform a test, and, therefore, represents a visual failsafe which reduces the chances of erroneous test results due to underdosing of a test strip.

The length and width of the window are shorter than the length and width of the capillary test chamber. The window is dimensioned and positioned so that it overlays the entire width of the working electrode and at least about 10% of the width of the counter or reference electrode of the biosensor test strip. Preferably, the area of the roof surrounding the window is colored in a way that provides good color contrast between the sample, as observed through the window, and the roof area surrounding the window for ease of identifying sufficient dosing of the strip.

The third new feature of the test strip is the inclusion of a notch, or multiple notches, located at the sample application port. A notch is created in both the first insulating substrate and the roof of the strip. These notches are dimensioned and positioned so that they overlay one another in the test strip. These notches reduce a phenomenon called “dose hesitation”. When a sample is added to the sample application port of a notchless strip, the sample can hesitate in its introduction into the capillary test chamber. This “dose hesitation” adds to the testing time. When the test strip includes a notch, dose hesitation is reduced. Further, including the notch in both the first insulating substrate and the roof makes it possible for the test sample to approach the sample application port from a wide variety of angles. The angle of approach for the test sample would be more limited if the notch were only in the roof.

Finally, the fourth new feature of the test strip is a reagent that includes polyethylene oxide from about 100 kilodaltons to about 900 kilodaltons mean molecular weight at concentrations from about 0.2% (weight:weight) to about 2% (weight:weight), which makes the dried reagent more hydrophilic and sturdier. With the inclusion of polyethylene oxide, the test reagent can more readily withstand mechanical punching during strip assembly and mechanical manipulation by the user of the test strip. Further, the dried reagent, which will include from about 1.75% (weight:weight) to about 17.5% (weight:weight) polyethylene oxide, can easily redissolve, or resuspend, when an aqueous test sample is added to the strip's test chamber.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1

is an exploded view of a preferred embodiment of the present invention.

FIG. 2

shows a fully assembled, preferred test strip.

FIGS. 3a–3i

represent a preferred method of making the inventive test strip.

FIG. 4

is a cross sectional view of the test strip of

FIG. 2

through

line

28—28.

FIG. 5

is a cross sectional view of the test strip of

FIG. 2

through

line

29—29.

FIG. 6

illustrates hypothetical calibration curves for different lots of test strips.

DESCRIPTION OF THE INVENTION

The components of a preferred embodiment of the present inventive biosensor are shown in

FIGS. 1

, 2, 4 and 5. The biosensor includes first

insulating substrate

1, which has

first surface

22 and

second surface

23.

Insulating substrate

1 may be made of any useful insulating material. Typically, plastics, such as vinyl polymers, polyimides, polyesters, and styrenics provide the electrical and structural properties which are desired. First

insulating substrate

1 further includes

indentation

notch

3, and

vent hole

4. Because the biosensor shown in

FIG. 1

is intended to be mass produced from rolls of material, necessitating the selection of a material which is sufficiently flexible for roll processing and at the same time sufficiently stiff to give a useful stiffness to the finished biosensor, a particularly preferred first

insulating substrate

1 is 7 mil thick MELINEX 329 plastic, a polyester available from ICI Films (3411 Silverside Road, PO Box 15391, Wilmington, Del. 19850).

As shown in

FIG. 1

, electrically

conductive tracks

5 and 6 are laid down onto

first surface

22 of first

insulating substrate

Track

5 may be a working electrode, made of electrically conducting materials such as palladium, platinum, gold, carbon, and titanium.

Track

6 may be a counter electrode, made of electrically conducting materials such as palladium, platinum, gold, silver, silver containing alloys, nickel-chrome alloys, carbon, titanium, and copper. Noble metals are preferred because they provide a more constant, reproducible electrode surface. Palladium is particularly preferred because it is one of the more difficult noble metals to oxidize and because it is a relatively inexpensive noble metal.

Preferably, electrically

conductive tracks

5 and 6 are deposited on an insulative backing, such as polyimide or polyester, to reduce the possibility of tearing the electrode material during handling and manufacturing of the test strip. An example of such conductive tracks is a palladium coating with a surface resistance of less than 5 ohms per square on UPILEX polyimide backing, available from Courtalds-Andus Performance Films in Canoga Park, Calif.

Electrically

conductive tracks

5 and 6 represent the electrodes of the biosensor test strip. These electrodes must be sufficiently separated so that the electrochemical events at one electrode do not interfere with the electrochemical events at the other electrode. The preferred distance between

electrodes

5 and 6 is about 1.2 millimeters (mm).

In the test strip shown in

FIG. 1

, electrically

conductive track

5 would be the working electrode, and electrically

conductive track

6 would be a counter electrode or reference electrode.

Track

6 would be a reference electrode if made of typical reference electrode materials, such as silver/silver chloride. In a preferred embodiment,

track

5 is a working electrode made of palladium, and

track

6 is a counter electrode that is also made of palladium and is substantially the same size as the working electrode.

Three electrode arrangements are also possible, wherein the strip includes an additional electrically conductive track located between

conductive track

6 and vent

hole

4. In a three electrode arrangement,

conductive track

5 would be a working electrode,

track

6 would be a counter electrode, and the third electrode between

track

6 and vent

hole

4 would be a reference electrode.

Overlapping

conductive tracks

5 and 6 is second

insulating substrate

7. Second insulating

substrate

7 is made of a similar, or preferably the same, material as first insulating

substrate

Substrate

7 has a

first surface

8 and a

second surface

Second surface

9 is affixed to the surface of

substrate

1 and

conductive tracks

5 and 6 by an adhesive, such as a hot melt glue. An example of such glue is DYNAPOL S-1358 glue, available from Hüls America, Inc., 220 Davidson Street, PO Box 6821, Somerset, N.J. 08873.

Substrate

7 also includes

first opening

10 and

second opening

11. First opening 10 exposes portions of

conductive tracks

5 and 6 for electrical connection with a meter, which measures some electrical property of a test sample after the test sample is mixed with the reagent of the test strip.

Second opening

11 exposes a different portion of

conductive tracks

5 and 6 for application of

test reagent

12 to those exposed surfaces of

tracks

5 and 6. (In

FIG. 1

, the entire width of

conductive tracks

5 and 6 are exposed by opening 11. However, it is also possible to expose only a portion of the width of

conductive track

6, which is either a counter electrode or a reference electrode, as long as at least about 10% of the width is exposed by opening 11.) Additionally, second insulating

substrate

7 includes

indentation

19, which coincides with

indentation

2 as shown in

FIG. 1

Test reagent

12 is a reagent that is specific for the test to be performed by the test strip.

Reagent

12 may be applied to the entire exposed surface area of

conductive tracks

5 and 6 in the area defined by

second opening

11. Other applications of

reagent

12 in this region are also possible. For example, if

conductive track

6 in this region of the strip has a reference electrode construction, such as silver/silver chloride, then test

reagent

12 may only need to cover the exposed area of working

electrode

5 in this region. Further, the entire exposed area of an electrode may not need to be covered with test reagent as long as a well defined and reproducible area of the electrode is covered with reagent.

Overlaying a portion of

first surface

8 and

second opening

11 is

roof

13.

Roof

13 includes

indentation

14 and

notch

15.

Indentation

14 and notch 15 are shaped and positioned so that they directly

overlay indentations

2 and 19, and

notch

Roof

13 may be made of a plastic material, such as a transparent or translucent polyester foil from about 2 mil to about 6 mil thickness.

Roof

13 has

first surface

16 and

second surface

17.

Second surface

17 of

roof

13 is affixed to

first surface

8 of second insulating

substrate

7 by a suitable adhesive, such as 3 M 9458 acrylic, available from 3M, Identification and Converter Systems Division, 3M Center, Building 220-7W-03, St. Paul, Minn. 55144.

Preferably,

roof

13 further includes transparent or

translucent window

18.

Window

18 is dimensioned and positioned so that when

roof

13 is affixed to second insulating

substrate

7, the window overlays the entire width of

conductive track

5 and at least about ten percent of the width of

conductive track

Second surface

17 of

roof

13, the edges of opening 11, and

first surface

22 of insulating substrate 1 (and

conductive tracks

5 and 6 affixed to

first surface

22 of substrate 1) define a capillary testing chamber. The length and width of this capillary chamber are defined by the length and width of

opening

11 and the height of the chamber is defined by the thickness of second

insulting substrate

A preferred test strip may be manufactured as shown by the process illustrated by

FIGS. 3a–3i

. A sheet of insulative substrate material 21 (

MELINEX

329, 7 mil thickness, available from ICI) is coated on one side with hotmelt adhesive (DYNAPOL S-1358, available from Hills). (

FIG. 3a

)

Sheet

21 is cut along

line

24, thereby forming first insulating

substrate

1, coated with adhesive on

first surface

22, and second insulating

substrate

7, coated with adhesive on

second surface

2. (

FIGS. 3b and 3c

)

First opening

10 and

second opening

11 are created in

substrate

7 by die punching. (

FIG. 3d

) Next, electrically

conductive tracks

5 and 6, made of palladium on Upilex backing (available from Courtalds-Andus Performance Films), are unspooled from reels precut to about 1.5 millimeters width and laid down on

surface

22 of

substrate

1 so that the Upilex backing is adjacent to surface 22.

Surface

9 of

substrate

7 is laid adjacent to surface 22 of

substrate

1 and to

conductive tracks

5 and 6, thereby forming the sandwich structure shown in

FIG. 3e

. This sandwich structure is heat sealed.

test reagent

12 is then dispensed into

opening

11 and dried. (

FIG. 3f

) After

reagent

12 is dried, vent

hole

4 is created by a die punch. (

FIG. 3g

) Next,

roof

13, which includes

hydrophilic coating

25 and

window

18, is laid down over

opening

11 in a manner such that

window

18 overlaps the entire width of

conductive track

5 and about one half of the width of

conductive track

Roof

13 is released from a release liner and adhesively affixed to

surface

8 as shown in

FIG. 3h

Finally, individual test strips are punched out by a die punch as shown in

FIG. 3i

. The die punch may punch out test strips with or without

notch

15. If

notch

15 is included, the preferred angle of the vertex is 105°. Other angles, such as from about 45° to about 105°, are also possible for

notch

15. Further, notch 15 may be a single notch or multiple notches.

As noted above,

test reagent

12 is dispensed into the area of the test strip defined by

cutout

11. In the manufacturing process described above, it is preferred to provide corona treatment of

opening

11 before

test reagent

12 is applied. The application of corona treatment serves to increase the surface energy of the portion of

surface

22 and

conductive tracks

5 and 6 exposed by opening 11, encouraging uniform spreading of

reagent

12, and to pre-clean the portion of

conductive tracks

5 and 6 exposed by opening 11. Pre-cleaning of

conductive tracks

5 and 6 has been found to significantly improve the performance of the test strip. Corona treatment may be applied at Watt densities ranging from about 20 to about 90 watts per centimeter per second (W/cm/s) with an arc gap of about 1 millimeter (0.040 inch).

In the preferred method, the corona treatment is applied in blanket form over the surfaces shown in

FIG. 3e

at the above described watt densities. The treatment is most effective if applied within 5 minutes of

reagent

12 application and is typically practiced within 45 seconds of

reagent

12 application.

It is advantageous to reduce the effects of corona treatment on

surface

8 in order to ensure that

reagent

12 will fully coalesce in

opening

11 and does not have a greater affinity for

surface

8 than for the portion of

surface

22 and

conductive tracks

5 and 6 exposed by opening 11. A corona dissipation process, which allows for the selective reduction of the effects of a blanket corona treatment process, is incorporated to reduce the effects of the treatment on areas of the web (the sheet of test strips being processed) outside of

opening

11. This corona dissipation process consists of applying, a thin film of deionized water such that the water contacts surface 8, but will not contact

openings

10 and 11. Application of the thin film of water, which is preferably from about 1.5 microns to about 3.0 microns thickness (about 9.1 grams of water per square meter), may be accomplished via wick pad, flexographic print, or other commercially available coating application methods. The thin film of water is then dried from the surface, using forced convection or infrared methods just prior to application of

reagent

12. The net effect of this treatment is that the surface energy of

surface

8 is effectively reduced to less than 62 dyne prior to the application of

reagent

12 while the surface of area within opening 11 is maintained at it's post corona treatment surface energy.

In the preferred embodiment,

test reagent

12 is formulated for the measurement of glucose in a human blood sample. A protocol for the preparation of a liter of a preferred glucose reagent utilizing the enzyme quinoprotein (pyrroloquinoline quinone (PQQ)-containing) glucose dehydrogenase and the redox mediator ferricyanide is shown immediately below. (Quinoprotein glucose dehydrogenase is Enzyme Commission No. 1.1.99.17.)

Step 1: Prepare a solution of NATROSOL in deionized water. This is accomplished by adding 0.45 grams (g) of NATROSOL-250M (a microcrystalline hydroxyethylcellulose available from Aqualon) to 414 g of deionzied water while stirring at a speed of no less than 250 revolutions per minute (rpm) for a period of no less than 30 minutes. Mixing is best accomplished with an overhead rotating impeller using a three or four bladed turbine type propeller. The selection of propeller size and configuration is largely based on the radius of the mixing vessel being used. The selected propeller will typically have a radius greater than 75% of the radius of the mixing vessel.
Step 2: To the solution from Step 1, 5.6 g of AVICEL RC-591F (a microcrystalline cellulose available from FMC Corp.) is dispersed by gradually adding this AVICEL to the solution while mixing at a speed of no less than 570 rpm for no less than 60 minutes.
Step 3: To the mixture from Step 2, 8.4 g polyethylene oxide (300 kilodalton mean molecular weight) is added gradually while mixing at a speed of no less than 690 rpm for a period of no less than 45 minutes.
Step 4: A buffer solution is prepared by adding 12.1 g of monobasic potassium phosphate (anhydrous) and 21.3 g of dibasic potassium phosphate (anhydrous) to 450 g of deionized water.
Step 5: A50 g aliquot of the buffer solution is removed from the preparation of Step 4. To this 50 g aliquot, 12.5 mg of coenzyme PQQ (available from Fluka) is added. This solution is stirred until the coenzyme is completely dissolved. (A magnetic stir bar and magnetic stir plate are preferred for enzyme preparation.)
Step 6: To the solution from Step 5, 1.21 million units of the apoenzyme of quinoprotein glucose dehydrogenase is added gradually while stirring at a low speed (less than 400 rpm on a magnetic stir plate) to prevent foaming. The resulting solution is mixed for no less than 2 hours to allow the association of the enzyme and coenzyme to stabilize, thereby resulting in a solution of quinoprotein glucose dehydrogenase.
Step 7: To the buffer solution from Step 4, 59.1 g of potassium ferricyanicle is added. Next, 6.2 g of sodium succinate is added. The resulting solution is mixed until all solutes are completely dissolved. After dissolution, the pH of the solution is assessed and is required to be approximately 6.76 plus or minus 0.05.
Step 8: The solution from Step 7 is gradually incorporated into the mixture from Step 3, while mixing at a rate of no less than 190 rpm.
Step 9: To the mixture from Step 8, 20 g trehalose is added, while mixing at a rate of no more than 190 rpm for a period of not less than 10 minutes.
Step 10 : 0.35 g of TRITON X-100 surfactant, available from Boehringer Mannheim Biochemicals, is added to the mixture from Step 9, while mixing at a rate of no more than 190 rpm. This mixture must continue mixing for no less than 5 minutes.
Step 11: The enzyme solution from Step 6 is added to the mixture from Step 10 and the now complete reagent is mixed at a rate of no less than 190 rpm for a period of no less than 30 minutes.
Step 12: The reagent can now be filtered, as needed by the manufacturing equipment, by passing it through a 100 micron sieve bag or through a 100 micron filter integral to a pumping system.

The apoenzyme of quinoprotein glucose dehygrogenase, specified above, is obtained from Boehringer Mannheim GmbH in Germany (Boehringer Marinheim GmbH identification number 1464221). Alternatively, this apoenzyme may be obtained from Acinetobacter Calcoaceticus by the following protocol, recited in Duine et al., FEBS Letters, vol. 108, no. 2, pps. 443–46.

Acinetobacter Calcoaceticus are grown on a mineral salt medium supplemented with 0.02 molar (M) sodium succinate or 0.10 M ethanol at 22° C. with good aeration. The cells are harvested at the end of the logarithmic phase and a wet-cell yield of ˜4 g/l can be obtained.

Frozen cells (10 g) are thawed and mixed with 15 milliliters (ml) of 36 millimolar (mM) Tris/39 mM glycine buffer. After adding 6 milligrams (mg) lysozyme, the suspension is stirred at room temperature for 15 min. and centrifuged for 10 min. at 48,000×g. The supernatant is discarded and the pellet extracted twice with 36 mM Tris/39 mM glycine buffer, containing 1% TRITON X-100 surfactant. The supernatants of the centrifugation steps are combined and used immediately.

The cell-free extract is added to a DEAE-Sephacel column (13×2.2 centimeters (cm)), equilibrated with 36 mM Tris/39 mM glycine buffer, containing 1% TRITON X-100 surfactant and the column is washed with the same buffer. The enzyme does not adhere to the column material and the combined active fractions are titrated with 2 M acetic acid to pH 6.0. This solution is added immediately to a column of CM-Sepharose CL-6 B (5×1 cm), equilibrated with 5 mM potassium phosphate (pH 6.0). After washing the column with the same buffer until no TRITON X-100 surfactant is present in the eluate, the enzyme is eluted with 0.1 M potassium phosphate (pH 7.0).

The enzyme is then dialyzed against 0.1 M sodium acetate (pH 4.5), containing 3 M potassium bromide at 4° C. for 72 hours. The enzyme is then dialyzed against 0.02 M potassium phosphate (pH 7.0) for 12 hours, resulting in the apoenzyme.

In the preferred test strip, opening 11 is about 3.2 millimeters by about 6.7 millimeters. In the preferred embodiment of a glucose test strip, 4.5 microliters of test reagent made by the above protocol is added to

opening

11. (See

FIG. 3f

) This amount of reagent will substantially cover the exposed surfaces of

conductive tracks

5 and 6 in

opening

11.

Test reagent

12 is then dried at about 70° C. for about 1 to 2 minutes.

The resulting, preferred, dried glucose reagent film will contain from about 2,000 to about 9,000 units of enzyme activity per gram of reagent. The preferred reagent will contain the following additional components per gram of reagent:

- 62.2 milligrams (mg) polyethylene oxide
- 3.3 mg NATROSOL 250 M
- 41.5 mg AVICEL RC-591 F
- 89.4 mg monobasic potassium phosphate
- 157.9 mg dibasic potassium phosphate
- 437.3 mg potassium ferricyanide
- 46.0 mg sodium succinate
- 148.0 mg trehalose
- 2.6 mg TRITON X-100 surfactant.

Importantly, including from about 0.2% by weight to about 2% by weight polyethylene oxide having a mean molecular weight from about 100 kilodaltons to about 900 kilodaltons, and preferably about 0.71% by weight polyethylene oxide having a mean molecular weight of 300 kilodaltons, in the wet reagent referred to above provides a test reagent that, when dried, is sturdier to strip processing steps, such as mechanical punching, sturdier to mechanical manipulation by test strip user, and that will redissolve or resuspend when an aqueous sample, such as human blood, is added to it. After drying, the percentage of polyethylene oxide ranges from about 1.75% (weight:weight) to about 17.5% (weight:weight). In the preferred, dried reagent, the percentage of polyethylene oxide is about 6.2% (weight:weight).

The preferred, dried, glucose reagent film thickness will be such that, in combination with the inherent properties of the test chemistry, the sensitivity of the test to interference from hematocrit variation is mitigated. In this preferred embodiment of the invention, the film thickness (as gauged by the ratio of wet reagent dispense volume to the surface area exposed by opening 11) is such that 4.5 microliters of reagent is dispensed into an area of approximately 22.5 square millimeters (the preferred area of opening 11). Including polyethylene oxide from about 100 kilodaltons to about 900 kilodaltons mean molecular weight in a film with the thickness described above, results in a sensor possessing a reduced sensitivity to hematocrit variation when glucose is measured from a human blood sample.

After

test reagent

12 is dried in

opening

11,

roof

13 is laid over

opening

11 and adhesively affixed to

surface

8 as described above.

Roof

13 itself is made in a separate process according to procedures described below.

Preferably,

roof

13 is made of MELINEX 561 polyester foil, having a thickness of 5 mil. A substantially opaque ink is printed on

first surface

16 in

pattern

27 such that

window

18 remains transparent or translucent. The window is positioned and dimensioned so that when the roof is affixed to

surface

8, it will align with opening 11 as shown in

FIG. 3h

second surface

17, an adhesive system is laminated in order that the roof may be ultimately affixed to

surface

8. This adhesive system can conveniently be an acrylic adhesive such as available from many commercial sources, but preferably part number 9458 from 3M Inc.

In addition, prior to placing the roof on

surface

8, a piece of coated transparent or translucent plastic, preferably a polyethylene terephthalate (PET), such as Melinex S plastic from about 0.001 to about 0.004 inch thick, is placed against the adhesive system on

second surface

17, and aligned with, and extending beyond the dimensions of

window

18. This coated plastic is

hydrophilic coating

25.

Coating

25 is specifically chosen to impart a hydrophilic nature to the internal surface of the capillary test chamber to encourage flow of an aqueous sample, such as blood, into the test chamber.

Coating

25 can be chosen from many available coatings designed to present a hydrophilic surface, but product number ARCARE 8586, available from Adhesives Research, Inc., is preferred.

Coating

25 also acts to prevent direct contact of the roof's adhesive to

reagent

12.

Finally,

roof

13 is placed onto

surface

8. (See

FIG. 3h

) It is at this stage that the transparent or

translucent window

18 defined by the absence of printed ink on

roof

13 must align with opening 11 as shown in

FIG. 3h

. The dimensions of transparent or

translucent window

18 should be chosen such that a substantial fraction of the width (greater than about 75%) of the underlying capillary channel is visible through

window

18. The orthogonal dimension of

window

18 should expose the entire width of the working

electrode

5. Therefore, when a sample, such as blood, is introduced into the capillary test chamber, through

sample application port

20, it is possible for a user of reasonable visual acuity to determine if the window is entirely full of the sample. By choosing the window dimensions as just stated it is possible to provide feedback for the user of the test strip that the strip has been sufficiently dosed with a test sample. Visual confirmation of the window being full provides assurance that a sufficient area of the working electrode is covered with sample and that a sufficient part of the counter or

reference electrode

6 is also covered. This coverage of the electrodes by the test sample is important to achieving an accurate test in a capillary-fill electrochemical biosensor. This visual confirmation of sufficient dosing of the test strip provides a safeguard against erroneous test results due to undetected underdosing of the test strip.

Completed

test strips

26 are used in conjunction with a meter capable of measuring some electrical property of the test sample after addition of the test sample to sample

application port

20. (See

FIG. 2

) The electrical property being measured may be, for example, electrical current, electrical potential, electrical charge, or impedance. An example of measuring changes in electrical potential to perform an analytical test is illustrated by U.S. Pat. No. 5,413,690, the disclosure of which is hereby incorporated by reference.

An example of measuring electrical current to perform an analytical test is illustrated by U.S. Pat. Nos. 5,288,636 and 5,508,171, the disclosures of which are hereby incorporated by reference.

In the preferred embodiment,

test strip

26 is connected to a meter, which includes a power source (a battery). Improvements in such meters and a biosensor system can be found in U.S. Pat. Nos. 4,999,632; 5,243,516; 5,366,609; 5,352,351; 5,405,511; and 5,438,271, the disclosures of which are hereby incorporated by reference.

Many analyte-containing fluids may be analyzed by the electrochemical test strip of the present invention. For example, analytes in human body fluids, such as whole blood, blood serum, urine and cerebrospinal fluid may be measured. Also, analytes found in fermentation products and in environmental substances, which potentially contain environmental contaminants, may be measured.

For determining the concentration of glucose in a human blood sample with the preferred test strip recited above, wherein

tracks

5 and 6 are palladium of substantially the same size and the glucose reagent is the reagent specified above, a blood sample may be added to

sample application port

20. The sample will be drawn into the test chamber by capillary action. Once inside the test chamber, the blood sample will mix with

test reagent

12. After an incubation period of some desired time, for example, 30 seconds, a potential difference will be applied by the power source of the meter between

tracks

5 and 6. In the preferred embodiment, the applied potential difference is 300 millivolts. Current may be measured at any time from 0.5 seconds to about 30 seconds after the potential difference of 300 millivolts is applied. The measured current may be correlated to the concentration of glucose in the blood sample.

The current measured during the assay of an analyte from a fluid sample may be correlated to the concentration of the analyte in the sample by application of an algorithm by the current measuring meter. The algorithm may be a simple one, as illustrated by the following example:
[Analyte]=Ci_7.5+d
wherein [Analyte] represents the concentration of the analyte in the sample (see

FIG. 6

), i_7.5is the current (in microamps) measured at 7.5 seconds after application of the potential difference applied between the electrodes, C is the slope of line 30 (

FIG. 6

), and d is the axis intercept (

FIG. 6

By making measurements with known concentrations of analyte, calibration curve 30 (

FIG. 6

) may be constructed. This calibration will be stored in the Read Only Memory (ROM) key of the meter and will be applicable to a particular lot of test strips.

Lines

31 and 32 in

FIG. 6

represent other hypothetical calibration curves for two other different lots of test strips. Calibration for these biosensor lots would generate slightly different values for C and d in the above algorithm.

In a preferred method for analysis of glucose from a sample of human whole blood, current measurements are made at 0.5 second intervals from 3 seconds to 9 seconds after the potential difference is applied between the electrodes. These current measurements are correlated to the concentration of glucose in the blood sample.

In this example of measuring glucose from a blood sample, current measurements are made at different times (from 3 seconds to 9 seconds after application of the potential difference), rather than at a single fixed time (as described above), and the resulting algorithm is more complex and may be represented by the following equation:
[Glucose]=C₁i₁+C₂i₂+C₃i₃+ . . . C_ni_n+d,
wherein i₁is the current measured at the first measurement time (3 seconds after application of the 300 millivolt potential difference), i₂is the current measured at the second measurement time (3.5 seconds after application of the 300 millivolt potential difference), i₃is the current measured at the third measurement time (4 seconds after application of the 300 millivolt potential difference), i_nis the current measured at the n^thmeasurement time (in this example, at the 13^thmeasurement time or 9 seconds after application of the 300 millivolt potential difference), C₁, C₂, C₃, and C_nare coefficients derived from a muiltivariate regression analysis technique, such as Principle Components Analysis or Partial Least Squares, and d is the regression intercept (in glucose concentration units).

Alternatively, the concentration of glucose in the sample being measured may be determined by integrating the curve generated by plotting current, i, versus measurement time over some time interval (for example, from 3 seconds to 9 seconds after application of the 300 millivolt potential difference), thereby obtaining the total charge transferred during the measurement period. The total charge transferred is directly proportional to the concentration of glucose in the sample being measured.

Further, the glucose concentration measurement may be corrected for differences between environmental temperature at the time of actual measurement and the environmental temperature at the time calibration was performed. For example, if the calibration curve for glucose measurement was constructed at an environmental temperature of 23° C., the glucose measurement is corrected by using the following equation:
[Glucose]_corrected=[Glucose]_measured×(1−K(T−23° C.)),
wherein T is the environmental temperature (in ° C.) at the time of the sample measurement and K is a constant derived from the following regression equation:
Y=K(T−23),
wherein

Y = [ Glucose ] measured ⁢ ⁢ at ⁢ ⁢ 23 ⁢ ° ⁢ ⁢ C . - [ Glucose ] measured ⁢ ⁢ at ⁢ ⁢ T° ⁢ ⁢ C . [ Glucose ] measured ⁢ ⁢ at ⁢ ⁢ T° ⁢ ⁢ C .

In order to calculate the value of K, each of a multiplicity of glucose concentrations is measured by the meter at various temperatures, T, and at 23° C. (the base case). Next, a linear regression of Y on T−23 is performed. The value of K is the slope of this regression.

Various features of the present invention may be incorporated into other electrochemical test strips, such as those disclosed in U.S. Pat. Nos. 5,120,420; 5,141,868; 5,437,999; 5,192,415; 5,264,103; and 5,575,895, the disclosures of which are hereby incorporated by reference.

Claims (69)

1. A test strip, having an indentation along an edge for tactile identification of a sample application port, said test strip comprising:

a first insulating substrate having first and second surfaces, an indentation along an edge and a vent hole;

at least two electrically conductive tracks affixed to the first surface of the first insulating substrate;

a second insulating substrate having first and second surfaces, an indentation along an edge, and first and second openings, the second surface being affixed to the conductive tracks and the first surface of the first insulating substrate, the first opening exposing a portion of the conductive tracks for electrical connection to a meter capable of measuring an electrical property, the second opening being located along said edge and exposing a different portion of the conductive tracks and the vent hole;

a test reagent overlaying at least a portion of the conductive tracks exposed by the second opening; and

a roof having first and second surfaces and an indentation along an edge, the second surface of the roof being affixed to the first surface of the second insulating substrate and positioned so that the second surface of the roof and the surface of the first insulating substrate form opposing walls of a capillary fill chamber with a sample application port at said edge of the second insulating substrate, wherein the second opening in the second insulating substrate and the indentations in the first insulating substrate, the second insulating substrate, and the roof are aligned to thereby provide for tactile identification of the sample application port.

2. The test strip of

claim 1

, wherein the second surface of the roof includes a hydrophilic coating.

3. The test strip of

claim 1

, wherein the test reagent includes

reaction components appropriate for performing a test and from about 1.75% by weight to about 17.5% by weight polyethylene oxide having a mean molecular weight from about 100 kilodaltons to about 900 kilodaltons,

wherein the reagent will redissolve or resuspend upon addition of an aqueous test sample to the reagent.

4. The test strip of

claim 1

, wherein the test reagent includes reaction components appropriate for performing a test, and a dissolvable or suspendable film forming mixture including from about 0.2% by weight to about 2% by weight polyethylene oxide having a mean molecular weight from about 100 kilodaltons to about 900 kilodaltons,

wherein the test reagent may be applied to the test strip in a wet form, may be subsequently dried, and then redissolved or resuspended upon addition of an aqueous test sample to the dried reagent.

5. The test strip of

claim 4

, wherein the second surface of the roof includes a hydrophilic coating.

6. The test strip of

claim 1

, wherein the roof has a solid transparent or translucent window, which is dimensioned and positioned so that the window overlays the entire width of the electrically conductive track that is closest to the indentation of the first insulating substrate and at least about ten percent of the width of the other electrically conductive track.

7. The test strip of

claim 6

, wherein the second surface of the roof includes a hydrophilic coating.

8. The test strip of

claim 6

, wherein the test reagent includes

wherein the reagent will redissolve or resuspend upon addition of an aqueous test sample to the reagent.

9. The test strip of

claim 6

wherein the test reagent may be applied to the test strip in a wet form, may be subsequently dried, and then redissolved or resuspended upon addition of an aqueous test sample to the dried reagent.

10. The test strip of

claim 9

, wherein the second surface of the roof includes a hydrophilic coating.

11. The test strip of

claim 1

, further comprising:

a first notch along the indentation in the first insulating substrate, and a notch along the indentation in the roof, both first and second notches being positioned so that they overlay one another.

12. The test strip of

claim 11

, wherein the second surface of the roof includes a hydrophilic coating.

13. The test strip of

claim 11

, wherein the test reagent includes

wherein the reagent will redissolve or resuspend upon addition of an aqueous test sample to the reagent.

14. The test strip of

claim 11

wherein the test reagent may be applied to the test strip in a wet form, may be subsequently dried, and then redissolved or resuspended upon addition of an aqueous test sample to the dried reagent.

15. The test strip of

claim 14

, wherein the second surface of the roof includes a hydrophilic coating.

16. The test strip of

claim 11

wherein the roof has a solid transparent or translucent window, which is dimensioned and positioned so that the window overlays the entire width of the electrically conductive track that is closest to the indentation of the first insulating substrate and at least about ten percent of the width of the other electrically conductive track.

17. The test strip of

claim 16

, wherein the second surface of the roof includes a hydrophilic coating.

18. The test strip of

claim 16

wherein the test reagent may be applied to the test strip in a wet form, may be subsequently dried, and then redissolved or resuspended upon addition of an aqueous test sample to the dried reagent.

19. The test strip of

claim 18

, wherein the second surface of the roof includes a hydrophilic coating.

20. The test strip of

claim 16

, wherein the test reagent includes reaction components appropriate for the test, and a dissolvable or suspendable film forming mixture including from about 0.2% weight to about 2% by weight polyethylene oxide having a mean molecular weight of 300 kilodaltons.

21. The test strip of

claim 20

, wherein the polyethylene oxide is about 0.71% by weight.

22. The test strip of

claim 16

, wherein the test reagent includes

wherein the reagent will redissolve or resuspend upon addition of an aqueous test sample to the reagent.

23. The test strip of

claim 22

, wherein the mean molecular weight of the polyethylene oxide is 300 kilodaltons.

24. The test strip of

claim 23

, wherein the amount of polyethylene oxide, in the reagent is about 6.2% by weight.

25. A test strip comprising:

a first insulating substrate having first and second surfaces, a notch along an edge, and a vent hole;

at least two electrically conductive tracks affixed to the first surface of the first insulating substrate;

a second insulating substrate having first and second surfaces and first and second openings, the second surface being affixed to the conductive tracks and the first surface of the first insulating substrate, the first opening exposing a portion of the conductive tracks for electrical connection to a meter capable of measuring an electrical property, the second opening being located along an edge of the second insulating substrate and exposing a different portion of the conductive tracks, the notch in the first insulating substrate, and the vent hole;

a test reagent overlaying at least a portion of the conductive tracks exposed by the second opening; and

a roof having first and second surfaces and a notch along an edge, the second surface of the roof being affixed to the first surface of the second insulating substrate and positioned so that 1) the second surface of the roof and the first surface of the first insulating substrate form opposing walls of a capillary fill chamber with a sample application port at said edge of the second insulating substrate, and 2) the notch in the roof overlays the notch in the first insulating substrate;

whereby the notch in the roof and the notch in the first insulating substrate will cause a liquid aqueous sample, when touched to the sample application port, to flow into the capillary chamber without significant hesitation.

26. A test strip, comprising:

a first insulating substrate having first and second surfaces and a vent hole;

at least two electrically conductive tracks affixed to the first surface of the first insulating substrate;

a test reagent overlaying at least a portion of the conductive tracks exposed by the second opening; and

a roof having first and second surfaces and a solid transparent or translucent window, the second surface of the roof being affixed to the first surface of the second insulating substrate and positioned so that it overlays the second opening of the second insulating substrate and so that the second surface of the roof and the first surface of the first insulating substrate form opposing walls of a capillary fill chamber with a sample application port at said edge of the second insulating substrate, and the transparent or translucent window being dimensioned and positioned so that the window extends from the sample application port, and overlays the entire width of one of the electrically conductive tracks and at least about ten percent of the width of the other electrically conductive track.

27. A test strip, having an indentation along an edge for tactile identification of a sample application port, said test strip comprising:

a first insulating substrate having first and second surfaces and an indentation along an edge;

at least two electrically conductive tracks affixed to the first surface of the first insulating substrate;

a second insulating substrate having first and second surfaces, an indentation along an edge and an opening, the second surface being affixed to the conductive tracks and the first surface of the first insulating substrate, the second insulating substrate configured to expose a portion of the conductive tracks for electrical connection to a meter capable of measuring an electrical property, the opening being located along said edge and exposing a different portion of the conductive tracks;

a test reagent overlaying at least a portion of the conductive tracks exposed by the opening;

a vent hole communicating with the capillary fill chamber;

wherein the opening in the second insulating substrate and the indentations in the first insulating substrate, the second insulating substrate, and the roof are aligned to thereby provide for tactile identification of the sample application port.

28. The test strip of

claim 27

29. The test strip of

claim 27

further comprising a first notch along the indentation of the first insulating substrate, and a notch along the indentation in the roof, both first and second notches being positioned so that they overlay one another.

30. The test strip of

claim 29

31. A test strip comprising:

a first insulating substrate having first and second surfaces and a notch along an edge;

at least two electrically conductive tracks affixed to the first surface of the first insulating substrate;

a second insulating substrate having first and second surfaces and an opening, the second surface being affixed to the conductive tracks and the first surface of the first insulating substrate, the second insulating substrate configured to expose a portion of the conductive tracks for electrical connection to a meter capable of measuring an electrical property, the opening being located along an edge of the second insulating substrate and exposing a different portion of the conductive tracks, aid overlaying the notch in the first insulating substrate;

a test reagent overlaying at least a portion of the conductive tracks exposed by the opening;

a vent hole communicating with the capillary fill chamber;

32. A test strip comprising:

a first insulating substrate having first and second surfaces;

at least two electrically conductive tracks affixed to the first surface of the first insulating substrate;

a test reagent overlaying at least a portion of the conductive tracks exposed by the opening;

a roof having first and second surfaces and a solid transparent or translucent window, the second surface of the roof being affixed to the first surface of the second insulating substrate and positioned so that it overlays the opening of the second insulating substrate and so that the second surface of the roof and the first surface of the first insulating substrate form opposing walls of a capillary fill chamber with a sample application port at said edge of the second insulating substrate, and the transparent or translucent window being dimensioned and positioned so that the window extends from the sample application port, and overlays the entire width of one of the electrically conductive tracks and at least about ten percent of the width of the other electrically conductive track; and

a vent hole communicating with the capillary fill chamber.

33. An electrochemical test strip for conducting testing for the concentration of glucose in a blood sample, comprising:

a strip body including an edge surface extending about the perimeter of said strip body, said strip body defining a capillary channel and a vent in fluid communication with the capillary channel, said strip body comprising a sample application port open at a location along the edge surface, the capillary channel extending from the sample application port to at least the vent;

at least working and counter electrodes spaced from each other and positioned within the capillary channel at a location spaced from the perimetric edge surface;

a test reagent adjacent at least the working electrode; and

visualization means associated with the capillary channel for enabling a user to visually identify when a sufficient amount of blood sample has been added to the capillary fill chamber to accurately perform a test, said visualization means including a solid, transparent or translucent viewing material extending from at least adjacent the sample application port and overlying at least a portion of the capillary channel including said working electrode and at least a portion of said counter electrode,

said visualization means further includes a fill line extending across the capillary channel at a location intermediate the length of the capillary channel at a position such that movement of the blood sample to the fill line indicates sufficient filling of the test strip for conducting a test.

34. The test strip of claim 33 in which said fill line is formed by an opaque portion overlying a portion of the capillary test chamber.

35. The test strip of claim 34 in which the fill line extends at a location between the working electrode and the vent.

36. The test strip of claim 35 in which said fill line is formed by an opaque portion overlying a portion of the capillary test chamber.

37. The test strip of claim 33 in which said strip body includes opposed sides of the capillary channel, the sides being parallel and extending in a straight line from the sample application port, and orthogonal to the perimetric edge surface, to at least one of the electrodes, the fill line extending across the capillary channel in an orientation orthogonal to the opposed sides of the capillary channel.

38. The test strip of claim 37 in which said strip body further includes opaque portions generally aligned with the opposed sides of the capillary channel from adjacent the sample application port to at least one of the electrodes.

39. The test strip of claim 38 in which the opaque portions are spaced apart to reveal greater than about 75% of the width of the capillary channel.

40. The test strip of claim 33 in which said strip body includes a first substrate, a second substrate and a roof, the second substrate being positioned intermediate the first substrate and the roof and including an opening, the opening of the second substrate together with the first substrate and the roof defining the capillary channel.

41. The test strip of claim 40 in which said test strip includes conductive tracks connected with said working and counter electrodes, the first substrate having first and second surfaces, the working and counter electrodes being affixed to the first surface of the first substrate, the second substrate having first and second surfaces and an opening, the second surface of the second substrate being affixed to the first surface of the first substrate, the second substrate configured to expose a portion of the conductive tracks for electrical connection to a meter capable of measuring an electrical property, the opening being located along a perimetric edge surface of the second substrate and exposing said electrodes, and a roof having first and second surfaces and including a solid, transparent or translucent viewing material, the second surface of the roof being affixed to the first surface of the second substrate and positioned so that it overlays the opening of the second substrate and so that the second surface of the roof and the first surface of the first substrate form opposing walls of the capillary channel, the transparent or translucent viewing material extending from at least adjacent to the sample application port and overlying the entire width of one of the electrodes and at least about ten percent of the width of the other electrode.

42. The test strip of claim 40 in which the second substrate defines opposed sides of the capillary channel, the sides being parallel and extending in a straight line from the sample application port, and orthogonal to the perimetric edge surface, to at least one of the electrodes.

43. The test strip of claim 42 in which said test strip further includes opaque portions generally aligned with the opposed sides of the capillary channel from adjacent the sample application port to at least one of the electrodes, the fill line extending across the capillary channel in an orientation orthogonal to the opposed sides of the capillary channel.

44. The test strip of claim 43 in which the opaque portions are defined by the roof.

45. The test strip of claim 40 in which the opening of the second substrate defines opposed sides of the capillary channel, said visualization means including opaque portions generally aligned with the opposed sides of the capillary channel extending from adjacent the sample application port to at least one of the electrodes, the opaque portions being located in the area adjacent the capillary channel, the opaque portions having a color which contrasts with the color of the sample as observed through the viewing material,

whereby a user is able to visually locate the sample within the capillary channel by observation through the viewing material and is able to determine when the sample has filled the capillary channel at least up to the fill line.

46. The test strip of claim 45 in which the opposed sides of the capillary channel are parallel and extend in a straight line from the sample application port, and orthogonal to the perimetric edge surface, to at least one of the electrodes, and the fill line extends across the capillary channel in an orientation orthogonal to the opposed sides of the capillary channel.

47. An electrochemical test strip for conducting testing for the concentration of an analyte in a blood sample, comprising:

at least working and counter electrodes spaced from each other and positioned within the capillary channel at a location spaced from the perimetric edge surface; and

a test reagent adjacent at least the working electrode;

a solid, transparent or translucent viewing material extending from at least adjacent the sample application port and overlying at least a portion of the capillary channel, said strip body defining a viewing area comprising a portion of the viewing material allowing continuous visualization of the capillary channel from a portion thereof at or generally adjacent the sample application port, up to and including said working electrode and at least a portion of said counter electrode,

the viewing area being positioned and dimensioned such that blood introduced to the capillary channel through the sample application port and filling the viewing area at least up to a portion of said counter electrode under the viewing area is required for the test strip to have a sufficient blood sample to conduct a test,

said strip body further including a fill line extending across the viewing area at a location intermediate the length of the capillary channel at a position such that movement of the blood sample to the fill line indicates sufficient filling of the test strip for conducting a test.

48. The test strip of claim 47 in which said fill line is formed by an opaque portion overlying a portion of the capillary test chamber.

49. The test strip of claim 48 in which the fill line extends at a location between the working electrode and the vent.

50. The test strip of claim 49 in which said fill line is formed by an opaque portion overlying a portion of the capillary test chamber.

51. The test strip of claim 47 in which said strip body includes opposed sides of the capillary channel, the sides being parallel and extending in a straight line from the sample application port, and orthogonal to the perimetric edge surface, to at least one of the electrodes, the fill line extending across the capillary channel in an orientation orthogonal to the opposed sides of the capillary channel.

52. The test strip of claim 51 in which said strip body further includes opaque portions generally aligned with the opposed sides of the capillary channel from adjacent the sample application port to at least one of the electrodes.

53. The test strip of claim 52 in which the opaque portions are spaced apart to reveal greater than about 75% of the width of the capillary channel.

54. The test strip of claim 47 in which said strip body includes a first substrate, a second substrate and a roof, the second substrate being positioned intermediate the first substrate and the roof and including an opening, the opening of the second substrate together with the first substrate and the roof defining the capillary channel.

55. The test strip of claim 54 in which said test strip includes conductive tracks connected with said working and counter electrodes, the first substrate having first and second surfaces, the working and counter electrodes being affixed to the first surface of the first substrate, the second substrate having first and second surfaces and an opening, the second surface of the second substrate being affixed to the first surface of the first substrate, the second substrate configured to expose a portion of the conductive tracks for electrical connection to a meter capable of measuring an electrical property, the opening being located along a perimetric edge surface of the second substrate and exposing said electrodes, and a roof having first and second surfaces and including a solid, transparent or translucent viewing material, the second surface of the roof being affixed to the first surface of the second substrate and positioned so that it overlays the opening of the second substrate and so that the second surface of the roof and the first surface of the first substrate form opposing walls of the capillary channel, the transparent or translucent viewing material extending from at least adjacent to the sample application port and overlying the entire width of one of the electrodes and at least about ten percent of the width of the other electrode.

56. The test strip of claim 54 in which the second substrate defines opposed sides of the capillary channel, the sides being parallel and extending in a straight line from the sample application port, and orthogonal to the perimetric edge surface, to at least one of the electrodes.

57. The test strip of claim 56 in which said test strip further includes opaque portions generally aligned with the opposed sides of the capillary channel from adjacent the sample application port to at least one of the electrodes, the fill line extending across the capillary channel in an orientation orthogonal to the opposed sides of the capillary channel.

58. The test strip of claim 57 in which the opaque portions are defined by the roof.

59. The test strip of claim 54 in which the opening of the second substrate defines opposed sides of the capillary channel, said visualization means including opaque portions generally aligned with the opposed sides of the capillary channel extending from adjacent the sample application port to at least one of the electrodes, the opaque portions being located in the area adjacent the capillary channel, the opaque portions having a color which contrasts with the color of the sample as observed through the viewing material,

60. The test strip of claim 59 in which the opposed sides of the capillary channel are parallel and extend in a straight line from the sample application port, and orthogonal to the perimetric edge surface, to at least one of the electrodes, and the fill line extends across the capillary channel in an orientation orthogonal to the opposed sides of the capillary channel.

61. An electrochemical test strip for conducting testing for the concentration of glucose in a blood sample, comprising:

a strip body including an edge surface extending about the perimeter of said strip body, said strip body defining a capillary channel and a vent in fluid communication with the capillary channel, said strip body comprising a sample application port open at a location along the perimetric edge surface, the capillary channel extending from the sample application port to at least the vent, said strip body further defining a test area along the capillary channel between the sample application port and the vent;

at least working and counter electrodes spaced from each other and positioned within the test area of the capillary channel at a location spaced from the perimetric edge surface;

a test reagent received within the test area of the capillary channel and adjacent at least the working electrode;

said strip body including a solid, transparent or translucent viewing material overlying at least a portion of the capillary channel, including from a portion thereof at or generally adjacent the sample application port continuously up to and including said working electrode and at least a portion of said counter electrode, the viewing material permitting visualization of the blood sample as it moves through the capillary channel to the test area;

said strip body further including opaque portions defining a fill area viewable through the viewing material, the fill area comprising an area of the capillary channel needed to be filled to conduct an accurate test; and

a fill line extending across the capillary channel at a location intermediate the length of the capillary channel at a position such that movement of the blood sample to the fill line indicates sufficient filling of the test strip for conducting a test,

wherein observation through the viewing material of the blood sample within the capillary channel up to said electrodes comprises confirmation of sufficient blood sample being introduced into the capillary channel to conduct a test.

62. The test strip of claim 61 in which said fill line is formed by an opaque portion overlying a portion of the capillary test chamber.

63. The test strip of claim 61 in which the fill line extends at a location between the working electrode and the vent.

64. The test strip of claim 63 in which said fill line is formed by an opaque portion overlying a portion of the capillary test chamber.

65. The test strip of claim 63 in which the fill line extends at a location between the test area and the vent.

66. The test strip of claim 65 in which said fill line is formed by an opaque portion overlying a portion of the capillary test chamber.

67. The test strip of claim 61 in which the opaque portions are sized and dimensioned such that the blood sample is required to fill up to the fill line the portion of the capillary channel viewable through the viewing material in order to have a sufficient amount of blood sample to conduct a test.

68. The test strip of claim 61 in which the opaque portions extend continuously in alignment with the opposed sides of the capillary channel from the perimetric edge surface to the electrodes.

69. The test strip of claim 61 in which the opaque portions are sized and dimensioned such that the blood sample is required to fill up to the fill line the portion of the capillary channel viewable through the viewing material in order to have a sufficient amount of blood sample to conduct a test.

US10/692,031 1997-12-05 2003-10-23 Electrochemical biosensor test strip Expired - Lifetime USRE42924E1 (en)

Priority Applications (1)

Application Number	Priority Date	Filing Date	Title
US10/692,031 USRE42924E1 (en)	1997-12-05	2003-10-23	Electrochemical biosensor test strip

Applications Claiming Priority (3)

Application Number	Priority Date	Filing Date	Title
US08/985,840 US5997817A (en)	1997-12-05	1997-12-05	Electrochemical biosensor test strip
US10/008,788 USRE42953E1 (en)	1997-12-05	2001-12-07	Electrochemical biosensor test strip
US10/692,031 USRE42924E1 (en)	1997-12-05	2003-10-23	Electrochemical biosensor test strip

Related Parent Applications (1)

Application Number	Title	Priority Date	Filing Date
US08/985,840 Reissue US5997817A (en)	1997-12-05	1997-12-05	Electrochemical biosensor test strip

Publications (1)

Publication Number	Publication Date
USRE42924E1 true USRE42924E1 (en)	2011-11-15

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ID=25531850

Family Applications (8)

Application Number	Title	Priority Date	Filing Date
US08/985,840 Ceased US5997817A (en)	1997-12-05	1997-12-05	Electrochemical biosensor test strip
US09/386,562 Expired - Lifetime US6254736B1 (en)	1997-12-05	1999-08-31	Method of selectively increasing the hydrophilicity of a web
US09/386,210 Expired - Lifetime US6270637B1 (en)	1997-12-05	1999-08-31	Electrochemical biosensor test strip
US10/008,788 Expired - Lifetime USRE42953E1 (en)	1997-12-05	2001-12-07	Electrochemical biosensor test strip
US10/409,721 Expired - Lifetime USRE41309E1 (en)	1997-12-05	2003-04-09	Electrochemical biosensor test strip
US10/692,031 Expired - Lifetime USRE42924E1 (en)	1997-12-05	2003-10-23	Electrochemical biosensor test strip
US10/693,305 Expired - Lifetime USRE42560E1 (en)	1997-12-05	2003-10-24	Electrochemical biosensor test strip
US13/301,334 Expired - Lifetime USRE43815E1 (en)	1997-12-05	2011-11-21	Electrochemical biosensor test strip

Family Applications Before (5)

Application Number	Title	Priority Date	Filing Date
US08/985,840 Ceased US5997817A (en)	1997-12-05	1997-12-05	Electrochemical biosensor test strip
US09/386,562 Expired - Lifetime US6254736B1 (en)	1997-12-05	1999-08-31	Method of selectively increasing the hydrophilicity of a web
US09/386,210 Expired - Lifetime US6270637B1 (en)	1997-12-05	1999-08-31	Electrochemical biosensor test strip
US10/008,788 Expired - Lifetime USRE42953E1 (en)	1997-12-05	2001-12-07	Electrochemical biosensor test strip
US10/409,721 Expired - Lifetime USRE41309E1 (en)	1997-12-05	2003-04-09	Electrochemical biosensor test strip

Family Applications After (2)

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US10/693,305 Expired - Lifetime USRE42560E1 (en)	1997-12-05	2003-10-24	Electrochemical biosensor test strip
US13/301,334 Expired - Lifetime USRE43815E1 (en)	1997-12-05	2011-11-21	Electrochemical biosensor test strip

Country Status (14)

Country	Link
US (8)	US5997817A (en)
EP (3)	EP1036320B1 (en)
JP (7)	JP3342477B2 (en)
CN (4)	CN100538352C (en)
AR (2)	AR017788A1 (en)
AU (1)	AU732761B2 (en)
BR (2)	BRPI9816332B8 (en)
CA (3)	CA2481195C (en)
DE (3)	DE29824204U1 (en)
ES (1)	ES2153808T3 (en)
NO (1)	NO328686B1 (en)
NZ (1)	NZ504527A (en)
TW (3)	TW200530582A (en)
WO (1)	WO1999030152A1 (en)

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CA2309280C (en)	2006-02-21
EP1577668A2 (en)	2005-09-21
AU1619499A (en)	1999-06-28
USRE42953E1 (en)	2011-11-22
JP2002277429A (en)	2002-09-25
BRPI9816332B8 (en)	2021-07-06
EP1577669A3 (en)	2005-10-12
NZ504527A (en)	2002-10-25
ES2153808T1 (en)	2001-03-16
ES2153808T3 (en)	2008-07-01
BRPI9816332A2 (en)	2011-09-20
CN1591013A (en)	2005-03-09
CN1280673A (en)	2001-01-17
JP3574113B2 (en)	2004-10-06
JP4045280B2 (en)	2008-02-13
USRE42560E1 (en)	2011-07-19
AU732761B2 (en)	2001-04-26
JP3342477B2 (en)	2002-11-11
US5997817A (en)	1999-12-07
NO20002844L (en)	2000-06-02
JP3689050B2 (en)	2005-08-31
CN1603821A (en)	2005-04-06
EP1036320B1 (en)	2008-03-12
JP2002303599A (en)	2002-10-18
TWI294521B (en)	2008-03-11
BRPI9815122B1 (en)	2016-08-30
CN100538352C (en)	2009-09-09
NO328686B1 (en)	2010-04-26
EP1577668A3 (en)	2006-02-01
JP2001526388A (en)	2001-12-18
AR017788A1 (en)	2001-10-24
JP2004279433A (en)	2004-10-07
JP2005201914A (en)	2005-07-28
BRPI9815122A (en)	2000-10-10
JP2002310973A (en)	2002-10-23
DE29824204U1 (en)	2000-09-21
JP3630665B2 (en)	2005-03-16
JP4175517B2 (en)	2008-11-05
EP1577668B1 (en)	2012-08-22
US6254736B1 (en)	2001-07-03
DE69839250D1 (en)	2008-04-24
WO1999030152A1 (en)	1999-06-17
CN1173180C (en)	2004-10-27
BRPI9815122B8 (en)	2021-07-06
CA2481195C (en)	2010-04-13
CN1598574A (en)	2005-03-23
CN1325914C (en)	2007-07-11
CN100476420C (en)	2009-04-08
JP2005233976A (en)	2005-09-02
CA2481193C (en)	2009-03-24
CA2309280A1 (en)	1999-06-17
EP1577669A2 (en)	2005-09-21
CA2481195A1 (en)	1999-06-17
NO20002844D0 (en)	2000-06-02
CA2481193A1 (en)	1999-06-17
US6270637B1 (en)	2001-08-07
AR068119A2 (en)	2009-11-04
DE1036320T1 (en)	2001-03-01
EP1036320A1 (en)	2000-09-20
TW200530582A (en)	2005-09-16
TW200530585A (en)	2005-09-16
USRE43815E1 (en)	2012-11-20
DE69839250T2 (en)	2009-07-16
USRE41309E1 (en)	2010-05-04
BRPI9816332B1 (en)	2016-07-19
EP1036320A4 (en)	2005-06-15

Publication	Publication Date	Title
USRE42924E1 (en)	2011-11-15	Electrochemical biosensor test strip
AU2002300887B2 (en)	2005-02-24	Improved electrochemical biosensor test strip
AU2006202499B2 (en)	2008-02-28	Improved electrochemical biosensor test strip
AU2005202280B2 (en)	2007-08-23	Improved electrochemical biosensor test strip
MXPA00005159A (en)	2001-09-07	Improved electrochemical biosensor test strip

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USRE42924E1 - Electrochemical biosensor test strip - Google Patents