KR20070002042A - Analyte test system for determining the concentration of an analyte in a physiological fluid - Google Patents

Abstract

This invention provides a device for determining the concentration of an analyte like glucose, cholesterol, free fatty acids, triglycerides, proteins, ketones, phenylalanine or enzymes, in a physiological fluid like blood, serum, plasma, saliva, urine, interstitial and/or intracellular fluid, the device having an integrated calibration and quality control system suitable for dry reagent test strips with a very small sample volume of about 0.5muL based on to a new sample distribution system. The production of the inventive analyte test element involves only a small number of uncomplicated production steps enabling an inexpensive production of the strips. ® KIPO & WIPO 2007

Description

ANALYTE TEST SYSTEM FOR DETERMINING THE CONCENTRATION OF AN ANALYTE IN A PHYSIOLOGICAL FLUID}

The present invention relates to the field of quantitative analysis of specimens, such as glucose, in physiological fluids such as, for example, blood. More specifically, the present invention relates to a test object test system, a test method, and a manufacturing method for quantitatively measuring a test object in physiological body fluids.

Analyzing the concentration of specimens in physiological samples plays a promising role in the diagnosis and treatment of various diseases. The test object of interest is one containing, in particular, glucose, cholesterol, free fatty acids, triglycerides, proteins, ketones, phenylalanine, enzymes, antibodies or peptides in blood, plasma, urine or saliva.

Generally, a physiological fluid sample, such as capillary blood, is embedded in a test strip to assess the concentration of the specimen. Test strips are usually used in conjunction with devices that measure some electrical property, such as current, if the strip is designed to detect electroactive compounds, and with devices that measure light reflection and / or light transmission if they are designed for photometry do. In systems involving optical detection techniques, a mixture of enzymes and colorants known as colorants is embedded in the test strip. The specimen contained in the physiological body fluid on the test strip reacts with the reagent to change the reflection or transmission to indicate the concentration of the specimen in the test sample.

For example, glucose is measured quantitatively by oxidation with gluconic acid using glucose oxidase. The reaction product, hydrogen peroxide, in conjunction with a peroxidase, such as horseradish peroxidase, converts a substrate, such as an indicator, into a detectable chromogenic product in proportion to the concentration of glucose in the sample solution.

Measuring glucose levels in whole blood samples is a very common test. Because diabetes causes dangerous physiological complications that lead to vision loss, kidney failure, and other serious medical consequences, strict treatment and disease management can be minimized with a combination of exercise, diet, and medication. Some patients have to test their blood glucose levels more than four times a day. Thus, as well as these patients, therapists and hospitals need an accurate, reliable and ideally inexpensive way to tailor treatment methods to avoid long-term complications of diabetes.

As awareness of diabetes has increased, self-monitoring and self-treatment has relied on the availability of appropriate devices, which has resulted in the development of numerous devices and methods for personal as well as testing purposes for treatment. Currently available devices include pregnancy, ovulation, coagulation, ketone and cholesterol tests (such as for non-consumable screening), but the most prominent in the field of self monitoring is also glucose testing of capillary blood.

An example of a device for monitoring the concentration of a specimen, such as glucose in the blood, is those disclosed in US Pat. No. 4,935,346. The method of this document involves reading the reflection value from an inert porous matrix surface containing a reagent that reacts with the specimen to generate a light absorption reaction product. Most prior art devices are designed to have one measuring band or measuring chamber, either directly introduced through a test sample or through a flow path or channel, which test chamber or test film is subjected to a reaction that causes a detectable color change of the sample liquid. Contains all necessary materials.

U.S. Patent 5,430,542 discloses a disposable optical cuvette and a method of making the same. This cuvette is equipped with two optically transparent liquid impermeable plastic sheets. A third adhesive sheet is sandwiched between the two transparent plastic sheets, and these three sheets are all compressed and sealed together.

U.S. Patent 5,268,146 discloses a qualitative test panel for testing specimen presence, containing all reagents and components necessary to visually indicate the presence of a specimen in a sample.

U.S. Patents 4,761,381 and 5,997,817 provide a device for placing a liquid sample to be analyzed into a sample application port and introducing the liquid into a capillary channel connected to a reaction chamber containing a substance capable of detecting a desired component of the liquid. It is starting.

US 2002/0110486 A1 and US 2003/0031594 A1 disclose a fluid medical diagnostic device capable of measuring the concentration of a specimen or the characteristics of biological fluids, in particular the clotting time of blood, which device is provided at one end. At the other end, a bladder is provided for application of the sample port for introduction into the measurement zone, where the physical parameters of the sample are measured and linked to the characteristics of the body fluid or the concentration of the specimen. .

When making these strips on a large scale, raw material and process variations do not guarantee proper strip-to-strip regeneration from one batch to the next. Therefore, it is necessary to assign a calibration code for each strip lot to compensate for these variations. The calibration code can be marked on the strip container and the user must enter the code into the meter if a new batch of strips is used. If the user fails to enter a new calibration code or enters an incorrect code, the measurement result will be incorrect. Several prior art strips, such as those disclosed in US Pat. No. 6,168,957 B1, are designed to incorporate a calibration code on the strip, so that the calibration code can be read before calculating the glucose concentration. This disposal characteristic of single-use diagnostic strips allows only destructive tests due to the consumption of reagents during the measurement phase, and thus the manufacturer can only statistically evaluate batch performance, thus ensuring 100% confidence in the performance of individual test strips. Can not.

More importantly, this type of calibration code conveys only retrospective information for an analytical strip-reading device or meter. Thus, the meter will not be able to evaluate the actual records of certain reagent test strips, such as incorrect storage conditions or incomplete packaging, such that the strips provide entirely false and off-scale readings compared to preliminary program data or assays. Doing so will cause an incorrect message.

The user can only check and verify the accuracy and function of the reagent test strips using specially prepared control solutions of known concentrations provided by the manufacturer. Another disadvantage is that such quality checks lead to increased costs because of increased strip consumption. Likewise, this method does not account for quality variations within a batch.

Some devices of the prior art have integrated positive and / or negative controls that are activated by sample addition. For example, in the case of U. S. Patent No. 5,268, 146 described above, preferred embodiments of the test device include a built-in positive control and / or a built-in negative control composed of additional measurement bands containing reagents, wherein the reagents are themselves visible to the indicator. It will prevent a change in phosphorus or occur regardless of the presence of the specimen in the test sample. In addition, the test device of US Pat. No. 4,578,358 for detecting the presence of occult blood in body material includes a positive and negative control band.

An integrated positive or negative control, as disclosed in the two patents described above, and commonly known from pregnancy tests, is useful only in conjunction with a qualitative and threshold test panel or strip that indicates the presence of a subject, but also quantitative measurement of a subject such as glucose in whole blood. Provides information that is meaningless in assuring quality.

In addition, the measurement process may be affected by other variable factors in the physiological sample solution. A typically troublesome problem in whole blood analysis is that due to the variability in erythrocyte levels it may not reflect the sample concentration of the sample.

In view of the above shortcomings, the object of the present invention is to allow the user to perform the test properly by compensating for and compensating for any variation that may occur due to variations in the production process or the variability of the test sample itself. It is to provide a device with an integrated calibration system that ensures that the results are accurate and reliable.

So far, dry reagent test strips with an integrated calibration system have not been disclosed in the prior art, except that the various literatures of the prior art have used a plurality of articles used to detect a plurality of specimens or to incorporate a positive or negative control as described above. It just describes a test strip with a response band of.

Particularly interesting prior art test strips are disclosed in US 2002 / 0110486A1 and US 2003 / 0031594A1, which have a plurality of reaction zones which are used for quality assurance purposes but which are not used for the internal calibration of the strip. Doing. This test strip requires about 20 μL of blood and is used to measure prothrombin time, an important variable in characterizing blood coagulation. However, if the user needs to test several times a day for proper management of diabetes, the need for a large amount of sample to this extent requires only about 1 μL of whole blood, but also allows the patient to perform the calibration process. Not only is it not practical compared to current glucose measurement systems, it is also disadvantageous.

Reducing the volume of cavities and channels that form the measurement cavities in the strips described here requires a complex and expensive production process, such as "micro-molding," which is not sufficient for large scale production of inexpensive disposable sensors. Inappropriate.

Thus, another object of the present invention is not only to require a small amount of physiological body fluids, but also to assist the self-monitoring of blood glucose or other important physiological variables, as the production process does not go through a complex production stage. A low-cost, test specimen system for dry reagent test strips is provided.

Summary of the Invention

The present invention relates to a test such as glucose, cholesterol, free fatty acids, triglycerides, proteins, ketones, phenylalanine or enzymes in physiological fluids such as blood, serum, plasma, saliva, urine, interstitial cells and / or intercellular fluids. By providing a device for measuring the concentration of water, the device has an integrated calibration and quality control system suitable for dry reagent test strips, even with very small amounts of samples of about 0.5 μL, based on the new sample distribution system. . Since only a few manufacturing processes are required to manufacture the analyte test element of the present invention, the production cost of the strip can be made low.

Due to this integrated calibration process, the specimen test system of the present invention provides reliable results, regardless of blood type, hematocrit concentration, temperature, and the like. In addition, production variations are offset by the integrated calibration process. In addition, it is possible to detect the aging of the active ingredient, so that it is possible to offset and / or report it, thus allowing long-term storage of the product under suitable storage conditions.

The present invention provides a specimen test member for measuring the concentration of one or more specimens in a physiological sample, which has a first surface and a second surface spaced by a predetermined distance from each other, wherein the two surfaces have a high surface energy. The two substantially identical pattern forming parts having a low surface energy with and are provided in a mostly aligned manner, thereby providing a sample dispensing system having two or more detection parts, wherein the applied physiological fluid is limited to the high surface energy part. do.

This sample dispensing system contained inside the test member has no mechanical and / or structural properties similar to walls, groves or channels for manipulating physiological body fluids or other aqueous samples. The specimen test member is described through several embodiments that are suitable for various calibration processes and are also acceptable for different specimens and chemical measurements; It can be easily integrated into test strips used in more complex arrangements, such as specimen test discs or bandoliers to provide a base unit for one-time or multiple measurements.

In a preferred embodiment, the specimen test member is

N predetermined detection portions of said first surface coated with a catalyst formulation that facilitates detection of the specimen in physiological fluids, and

Detecting n number of said second surface coated with a, m of the blank formulation and the calibration compound of n calibration formulations made of nm of the formulation having a concentration difference of the formed close to the center of the analyte test element (here, n Is an integer greater than 2, m is an integer of 1 or more, where n > m )

Wherein the detecting means obtains n results from 2 n predetermined detection parts and then the processing means calculates nm correction coefficients according to the following polynomial calibration equation,

One regression coefficient that verifies the quality of the calculated nm calibration coefficients of the calibration equation, and the unknown concentration of the specimen in the physiological fluid sample.

In another aspect, the present invention provides a method of manufacturing an object test member of the present invention, which method includes the following processes:

A process for generating a high surface energy portion and a low surface energy portion in a base layer having a first surface, wherein the high surface energy portion has n predetermined (where n is an integer greater than 2) hydrophilic pathways ) To form,

Generating a corresponding pattern of the high surface energy portion and the negative surface energy portion on the cover layer having the second surface,

Coating a catalyst formulation on the n detection portions of the first surface, wherein the catalyst formulation facilitates the detection of the concentration of the specimen contained in the physiological bodily fluid sample using transmission or absorption photometry;

The step of coating with the n calibration formulations on n detection of the second surface, said n calibration formulations made up of n -m are two formulations having different of m blank formulations and the calibration compound concentrations, where m Is an integer greater than or equal to 1 and n > m , wherein the calibration compound is the same or substantially equivalent to the test subject, so that the same chemical reaction as the test subject in the physiological bodily fluid sample can be induced in the catalyst formulation,

Stacking the layers of the first and second surfaces on opposite sides of the center layer having discontinuities that provide cavities to the sample distribution system formed by the high surface energy portions on the first and second surfaces of the base and cover layers. fair,

Process of punching or cutting the laminated sheets into final shape

It is made, including.

Other features, advantages and preferred embodiments of the invention will become apparent from the following detailed description with reference to the accompanying drawings.

1 and 2, the test strip 1 of the present invention includes a base layer 2, a center layer 3 covering the base layer, and a cover layer 4 covering the center layer 3. Has a batch structure. Center layer 3 provides a discontinuity 5 that creates a hollow cavity in conjunction with base layer 2 and cover layer 4. Within the cavity is a sample dispensing system 6, which is connected to a sample application section 9 located on one side of the test strip. In order to apply the sample more easily, the sample applying unit 9, which is an interface to the user, preferably has a shape of curve 10 protruding convexly from one main side of the test strip. The vent 11 is located on the second main side of the test strip, opposite the sample application section 9, 10, and exhausts air while the physiological fluid is distributed to the predetermined detection sections 6a, 6'a (Fig. 3). Reference). Here, it can be seen that this structure requires only one vent regardless of the area of the predetermined inspection portion used in the specimen test member. The aforementioned member of the sample dispensing system having a high surface energy portion, a sample applying portion, a vent, a center layer, and a discontinuity in the center layer forms the entire specimen test member, and causes an intrinsic capillary action to produce an applied physiological fluid. Distribute it to the detector.

In addition, specimen test strip 1 includes registration features 7, 8, which are useful for distinguishing different types of specimen test strips for measuring different specimens. By this means, it is possible to instruct a plurality of specimen meters for carrying out a special program or process with the identifiable parameters in the strip insertion required for measuring a particular specimen. As shown in FIG. 3, which schematically illustrates the multi-layer arrangement of FIGS. 1 and 2, base layer 2 provides a first surface 2a and cover layer 4 provides a second surface 4a. The first surface 2a and the second surface 4a are patterned with the areas that will create the sample distribution system 6. The pattern of the sample dispensing system 6 includes a predetermined number of specimen detecting sections 6a and sample paths 6b, which are woven so that they are mostly aligned in a multi-layer batch assembly. The center layer 3 defines the distance between the first surface 2a of the base layer 2 and the second surface 4a of the cover layer 4 and has a discontinuity 5 so that the first surface 2a of the base layer 2 and the second surface of the cover layer 4 Together with 4a, a hollow cavity is formed. The sample distribution system 6 to be formed between the first surface 2a and the second surface 4a is formed in a cavity made by the discontinuity 5 of the center layer 3 and the second surface 4a of the cover layer 4 and the first surface 2a of the base layer 2. Located. Preferably, this hollow cavity is preferably designed to be substantially larger than the sample dispensing system.

Since the sole purpose of the discontinuity 5 of the center layer is to form a cavity for the sample dispensing system 6, the discontinuity 5 of the center layer 3 may have various shapes, an example of which is shown in FIG. FIG. 4A shows an umbrella-shaped test member cavity 12, FIG. 4B shows a rectangular test member cavity 13, and FIG. 4C shows a circular cavity 14. Since the discontinuity 5 of the center layer 3 does not affect the size of the predetermined detection portion 6a and the size of the path 6b of the sample distribution system 6, it does not affect or change the required sample volume. Compared to the sample dispensing system 6, the shape of the cavity shown in FIG. 4 is rather simple, allowing a simple drilling tool to work quickly without requiring high registration accuracy.

The sample distribution system 6 located in the cavity formed by the discontinuities 5 of the center layer 3 and the first surface 2a of the base layer 2 and the second surface 4a of the cover layer 4 has a high surface energy portion and a low surface area on the surfaces 2a and 4a. It is formed by creating a surface energy portion. The high surface energy portion and the low surface energy portion on the first surface 2a of the base layer 2 and the second surface 4a of the cover layer 4 are aligned so that they are mostly aligned with each other. Since the applied physiological fluid or other aqueous sample only wets the high surface energy portion, it is limited to only between the predetermined flow path 6b of the sample distribution system 6 and the detection portion 6a and the first surface 2a of the base layer 2 and the second surface 4a of the cover layer 4. do.

FIG. 5 illustrates the structure of sample dispensing system 6 using hydrophobic “guiding elements”. In this embodiment of the specimen test member of the present invention, the base layer 2 and the cover layer 4 are coated with the hydrophobic layer 16, except for the portion where the sample flow and the detection portion are to be formed. The hydrophobic layer 16 produces a low surface energy portion, which exerts a repulsive force on the applied sample liquid, thereby preventing the sample from entering the high energy portion to form the sample distribution system.

Preferably, hydrophobic layer 16 is preferably applied on a hydrophilic surface, which can be wetted by physiological body fluids and transmits light, in particular UV, near infrared and / or visible portions of the electromagnetic spectrum. The process described above requires a hydrophilic surface, which can be made from natural hydrophilic polymers such as cellophane or glass, as well as from the hydrophobic surfaces of common polymers (illustrated below). For example, by making it hydrophilic by using a physicochemical plasma deposition or coating process of a hydrophilic monomer that can be vacuum deposited in ethylene oxide, ethylene glycol, pyrrole or acrylic acid. Subsequently, the pattern of "pottery rich" can be performed by printing hydrophobic ink on the hydrophilic surfaces of the base layer and the cover layer.

Suitable hydrophobic inks have a contact angle with water, usually greater than 70 °, with a surface energy of typically less than 33 mN / m, and systems having isooctyl acrylate, dodecyl acrylate, styrene derivatives or partially fluorinated carbon chains. As such, it contains a monomer having a hydrophobic functional group, an oligomer, and a prepolymer.

6 shows yet another structure of a sample distribution system using a hydrophilic pathway. In this test specimen specific example, the base layer 2 and the cover layer 4 are coated with a hydrophilic layer 17 to form a high surface energy layer.

Hydrophilic reagents printed on hydrophobic surfaces can be very well wetted by physiological fluids; Thus, the high surface energy portion, which makes the hydrophilic path of the sample distribution system, exert a strong capillary force on the applied physiological sample, and deliver the sample liquid to the spaced detector.

Hydrophilic patterns can be made by printing cross-linked and / or partially insoluble hydrophilic or amphiphilic reagents on hydrophobic surfaces. Inks having hydrophilic functional groups can be demonstrated by extensive selection of crosslinkable water soluble polymers, and particularly useful are acrylate derivatives made from polyalcohols, polyethylene glycols, polyethylene oxides, vinylpyrrolidone, alkyl phosphocholine and other materials. to be; Particularly useful ones include organo-modified silicone acrylates, which are crosslinkable species of organo-modified polysiloxanes. Contact angles with water by suitable coatings are usually less than 25 ° and typical surface energies are in excess of 70 mN / m.

Base layers 2 and cover layers 4 suitable as substrates in the printing process are polyesters and polyester resins containing glass, polyvinyl acetate, poly-methyl-methacrylate, poly-dimethyl-siloxane, fluorene rings, polycarbonates and Polycarbonate-polystyrene graft copolymers, terminally modified polycarbonates, polyolefins, cycloolefins and cycloolefin copolymers, and / or olefin-maleimide copolymers.

If such substrates have intermediate hydrophobic properties, it is also possible to print the hydrophilic pathway in a surrounding hydrophobic pattern, ie the combination of the structures of FIGS. 5 and 6.

In another embodiment, a sample distribution system is provided by providing a hydrophilic / hydrophobic pattern to only one of the first and second surfaces. Preferably, however, it is desirable to provide an even pattern of high surface energy and low surface energy for both the first and second surfaces to ensure proper distribution of the sample liquid within the hydrophilic path of the sample distribution system.

In addition, it is possible to physically raise the high surface energy portion of the first and second surfaces from the low surface energy portion by etching or embossing.

The volume required in the sample dispensing system contained in the specimen test member of the preferred embodiment is approximately 0.5 μL-1.0 μL, which is very small and only requires 100 nL-150 nL per detector, high surface energy and low surface energy. It is irrelevant whether it is made by additional hydrophobic conductors, hydrophilic pathways, or a combination of both. However, one of ordinary skill in the art will readily appreciate that the capacity of the sample dispensing system may vary depending on the various designs or the number of predetermined detectors used.

As mentioned above, with the test strip technique according to the prior art, it is very difficult or even impossible to manufacture a sample dispensing system having a plurality of sample dispensing paths and a detection section and operating at a very small volume of the above-mentioned degree. The amount of physiological sample required for measurement in the test member of the present invention is only 1/40 of the amount required for device operation as disclosed in, for example, US 2002/0110486 A1 and US 2003/0031594 A1 to Shartel et al., Only about one tenth of the capacity required by prior art micro-cube systems (HemoCue Glucose Systems).

FIG. 7 shows another pattern of a sample dispensing system that may be realized by the hydrophilic pathway shown in FIG. 6 or a hydrophobic “conductor member” as shown in FIG. 5, or a combination of the hydrophilic pathway and a hydrophobic ceramic member. will be. The cell AI of FIG. 7 illustrates all cases of the simplest sample dispensing system. Column A of FIG. 7 is the official design of the sample dispensing system, without background correction, while column B provides the design of the sample dispensing system, which provides background correction. Column C indicates the highest order of polynomial calibration equations achievable with adjacent designs, and column n indicates the required number of predetermined detectors on each surface, indicating the number of measurements required. Letters written for each design indicate background calibration (c), sample (1), and all relevant calibrations (2, 3, 4, 5, 6) with increasing amounts of calibration compound. The simplest calibration is represented by a linear equation where the relationship between the measured value and the sample concentration is directly proportional. Calibration of specimen test members is generally performed using standard addition methods in which calibration compounds of different calibration portions are added to the sample and then calculated with a linear or monotone nonlinear calibration equation. 9 illustrates the case of I in more detail. The calibration model or order (column C) must be appropriate for the selected specimen and the detection chemistry used, so that the linear calibration model is not applicable to chemical reactions following the fourth-order model and vice versa. to be. However, it is still possible to use specimen test members planned for 5 linear additions for linear calibration, and the higher the volume, the more accurate the measurement and the quantitative verification, so that the correlation coefficient of the test compared to the linear calibration based on the 2 standard This will provide statistically significant results in terms of standard deviations and standard errors.

In addition, it is also possible to repeat sample and label measurements, so that it is possible to perform two independent linear calibrations on one particular sample of physiological fluid using the embodiment shown in column IV. Similarly, in measuring two specimens, it is possible to use the same specimen test member.

On the contrary, if the selected detection chemistry does not cause any interference problem, so if the reaction extracts and products of one reaction do not participate in another reaction and the indicator dye produced absorbs light in substantially different wavelength ranges, Multiple inspection systems can be realized within a predetermined detection part of the set.

In FIG. 3, the test strip detecting portion 6 ′ a of the sample distribution system 6 of the first surface 2 a of the base layer 2 is coated with the catalyst formulation 18, as shown in FIGS. 5 and 6. Catalyst formulation 18 is a reaction component that, if necessary, generates a promoter and an optically detectable product that performs a catalytic or noncatalytic reaction with the test specimen in conjunction with a promoter, thereby allowing the sample to It contains an indicator that makes it possible to detect a test object contained in the blood.

Preferably, the promoter is an enzyme selected from dehydrogenase, kinase, oxidase, phosphatase, reductase, and / or transferase. Any cocatalyst contained in the catalyst formulation requires the enzyme to facilitate a particular enzymatic reaction, such as glucose dehydrogenase, which requires 3-nicotinamide adenine dinucleotide.

In one assay system for measuring glucose concentration, glucose in the sample is oxidized by oxygen and glucose oxidase to form gluconic acid and H ₂ O ₂ . The amount of H ₂ O ₂ produced is then quantitatively determined by the following scheme (1) or scheme (2):

산화된 염료 (유색) + H₂ODye (colorless) + H ₂ O ₂

Oxidized Dye (Colored) + H ₂ O

H ₂ O ₂ + Fe ²⁺ → Fe ³⁺ + H ₂ O

Fe ³⁺ + dyes → Fe ³⁺ dye complex

In Scheme (1), peroxidase enzymes (eg horseradish peroxidase, microperoxidase) catalyze the oxidation of the dye to convert H ₂ O ₂ to H ₂ O. Color intensity is directly proportional to the concentration of glucose in the sample. Representative examples of the dyes include o-dianicidine, 4-amino antipyrine, and 3,3 ', 5,5'-tetramethibenzidine. In Scheme (2), H ₂ O ₂ oxidizes Fe ²⁺ to Fe ³⁺ . Fe ³⁺ forms colored chelate complexes at specific absorption peaks. Representative examples of chelate-dyes include xylenol oranges. The amount of Fe ³⁺ chelate complex formed is proportional to the amount of glucose in the sample.

In another assay used to measure glucose concentration in physiological fluids, Scheme (3) is used; Here glucose dehydrogenase (GDH) reacts specifically with glucose in the sample in the presence of the coenzyme (3-nicotinamide adenine dinucleotide (3-NAD)) to form NADH, the reduced form of 3-NAD. NADH is then reacted with an electron acceptor dye such as 3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide (MTT) (catalyzed by diaphorase enzyme) The color turns reddish dark purple (purple-redish). The color intensity measured at 640 nm is directly proportional to the glucose concentration in the sample.

Another strain of wild-type GDH is glucose dehydrogenase pyrroloquinolinquinone (GDH-PQQ), often used for electrochemical blood glucose testing, similar to chelate complex indicators or MTT as shown later in Scheme (2). It can be used for the optical detection method using the indicator dye formed by reduction.

Catalyst Formulation 18 may contain glucose oxidase or glucose dehydrogenase if the device is for measuring glucose concentration in physiological body fluids.

Thus, the enzymes contained in the catalyst formulations include cholesterol oxidase if the specimen is cholesterol; Alcohol oxidase if the test object is alcohol; If the test object is lactate, lactate oxidase may be used.

Indicators suitable for generating scientifically detectable products, alone or in combination with other chemicals, and in conjunction with suitable promoters such as enzymes, are preferably aromatic amines, aromatic alcohols, azines, benzidines, hydrazones, aminos. It is preferred to select among antipyrines, conjugated amines, conjugated alcohols and aromatic and aliphatic aldehydes. Specific examples of indicators include 3-methyl-2-benzonthiazolinone hydrazone hydrochloride; 3-methyl-2- (sulfonyl) -benzothiazolone hydrazone hydrochloride (MBTH); 8-amino-1-naphthol-5,7-disulfonic acid (chicago acid); 3,3 ', 5,5'-tetramethylbenzidine (TMB); 4,5-dihydroxy-2,7-naphthalene-disulfonic acid; 1-hydroxy-2-naphthalene-sulfonic acid; N, N-dimethyl-aniline; o-tolidine; 3-dimethyl-aminobenzoic acid (DMAB); 2,2-azino-bis93-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS); 3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide (MTT); And / or 3,5-dichloro-2-hydroxybenzene-sulfonic acid. If the chemical is an acid, water soluble salts of that acid, such as ammonium salts, may be used in the preparation of the catalyst formulation.

Various enzyme inks suitable as catalyst formulations are known in the prior art and include, for example, Wong et al. (US Pat. No. 6,312,888), Philips et al. (US Pat. No. 4,935,346), and Berti et al. (US Pat. No. 4,247,297). Suitable catalyst formulations in the present invention are based on enzymes or enzyme combinations, non-reactive bases, and indicator components (dyes) as promoters. The non-reactive base provides a carrier, which should be suitable for coating processes, preferably ink jet printing, enzyme stabilization and enzyme immobilization and indicator systems for the detector surface. An exemplary composition of a 100 mL formulation is shown below.

Non-Reactive Base:

65 ml of distilled water

Citric acid 2.4 g buffer system

Sodium citrate, 2H ₂ O 3.2 g buffer system

Polyethylene Glycol 1.0 g Crust Inhibitor

N-methylpyrrolidone 2.0 g for some indicator dyes

Cosolvent (optional ingredient)

BAS 3.0 g Enzyme Stabilization

Gafquat 440 (ISP) 1.0 mL film former

Advantage S (ISP) 1.0 g film former

PVA (low molecular weight) 1.5 g enzyme stabilization

Adjust pH to 6.5 and fill up to 100 mL

Catalyst Formulations:

(Add all ingredients to 100 mL non-reactive base)

GOD (Asperganes Niger) 2.0 g (250 U / mL)

POD (horse radish) 2.0 g (250 U / mL)

Indicator system a) TMB 0.801 g

Indicator system b) 0.915 g ABTS

Indicator system c) MBTH 0.719 g

DMBA 0.551 g

Indicator system d) MBTH 0.359 g

1.064 g from Chicago

Catalytic formulations can be formulated with indicator systems a) to d) in combination with various hydrogen peroxide producing enzymes such as GOD. Nevertheless, the pH of non-reactive base formulations needs to be adjusted to the requirements of new enzymes when GOD is replaced by another catalyst.

Examples of non-enzymatic catalysis include albumin detection using tetrabromphenol blue and ketone detection using phosphate buffered mixtures of nitroprusside and glycine in the visible part of the electromagnetic spectrum.

If the specimen test member is intended to be measured n times (where n is an integer greater than 2), the n detector 6'a on the first surface is coated with a catalyst formulation 18 to facilitate detection of the specimen in the physiological sample.

Referring to Figures 3, 5, and 6, the detector 6a of the second surface 4a of the cover layer 4 is characterized by being coated with a calibration formulation 19 containing a calibration compound.

Preferably, this calibration compound contained in calibration formulation 19 coated on the predetermined detection section 6a of the second surface 4a is the same or substantially equivalent to the test specimen, catalyzing the same chemical reaction as the test specimen in the physiological bodily fluid sample. It is good to be able to cause it during formulation. If the desired test in the physiological sample is glucose, the calibration compound is also preferably glucose.

Non-reactive bases as described for the catalyst formulations are also suitable as calibration formulations and only need to add the calibration compound of the required concentration. N-methylpyrrolidone, a cosolvent necessary for some indicator dyes, may be omitted.

Accurate dosages of calibration compounds applied to different different detections are not only necessary for proper calibration, but also for the reliable calculation of the concentration of the specimen in the sample liquid. As EK, it is preferable that, as a catalyst formulation, a calibration formulation is also coated on a predetermined detection section by ink jet printing. By this means, the correct amount of calibration compound can be applied to a specific detection part.

If the analyte test element is designed to perform n meeting measured (where n is an integer greater than 2), the second surface 4a on the n predetermined detection, a calibration compound or analyte to different concentrations n - It is coated with m pieces of m blank formulations and formulation of n calibration formulations consisting of (wherein m is an integer of 1 or more, n> m). That is, at least one of the n detection sections of the sample distribution system does not contain a calibration compound.

If the physiological fluid is applied to the sample detection distributed to the predetermined detection by capillary action, the physiological fluid is the first on the one surface 2a second surfaces n as well as the catalytic formulation on the two predetermined detection of 4a n predetermined detection n The dog's calibration formulation is dissolved to form a mixture of the test article, the calibration compound (which may be an additional test article), a promoter, and an indicator dye. The optical density in these n mixtures varies proportionally with different levels of calibration compound plus unknown concentrations, allowing transmission and / or absorption photometry to yield n results of optical measurements and specimen concentrations. Become. Preferably, the catalyst formulation and calibration formulation applied to a given detector are readily dissolved in physiological body fluids and / or water so as to be in close proximity to each other to enable rapid diffusion mixing of both components, thereby providing all components contained in the detector. It is good to make them react quickly and to be able to measure the density | concentration of a test object quickly by the photometry method.

FIG. 8 is a diagram illustrating an arrangement of a detector for measuring the optical density of a sample in the test member according to FIG. 6. This layout includes a light source 20 that emits light 24 of a particular wavelength toward the sample detector. Light emitted from the light source 20 passes through the optical device 21, such as a diffuser or lens, and lens aperture 22, base layer 2, sample 15 and cover layer 4 of the detection section and is detected on the opposite side of the device by the detector means 23.

Since there are three or more detection units in the sample distribution system, and at least two of the detection units contain various known concentrations of the calibration compound, the computing means determines the unknown concentration of the specimen from n measurements performed on physiological fluids in the absence of the specimen test. It is possible to calculate.

9 is a diagram illustrating the calculation of specimen concentration in a sample by a linear standard addition method, which is a known calibration technique that has been used in various fields of analytical chemistry, but is first dried (in the present invention). It is used integrally for reagent test strips. In this embodiment, the sample dispensing system includes three specimen detection sections, two of which are coated with two predetermined calibration compounds of varying concentrations. After the physiological fluid is applied to the sample distribution system, a catalytic reaction is caused in the specimen detection unit, and the first absorbance value 25a of the sample located in the detection unit is measured for the first concentration of the calibration compound using a light emitter and a meter. The detection value of this detector shows a signal proportional to the sum of the concentration of the test specimen and the concentration of the first calibration compound. In the same vein, the second absorbance value 26a of the sample located in the detector relative to the second concentration of the calibration compound is measured, which represents a signal proportional to the sum of the concentration of the test specimen and the concentration of the second calibration compound. Furthermore, the 3rd light absorption value 27a of the detection part containing only a sample and the test material of unknown density is measured.

Since there is a linear correlation between the concentration of the specimen and the optical density according to Lambert-Beer's law, the computational means of the specimen test system calculates the coefficients for the calibration equation y = c ₀ + c ₁ x in the above example. Can be calculated by regression analysis. The concentration of the specimen in the physiological body fluid sample is obtained by 28 (0 = 28) of the calibration equation calculated above.

The general formula of the applicable calibration equation can be expressed as:

Scanning 2

Y = f (optical measurement result) in the formula; x = f (concentration of calibration compound); n = number of measurements.

This polynomial equation, in conjunction with the n value presented in FIG. 7, provides an example of the calibration model most useful for the various designs of the sample distribution system in the preceding figures. The y and x values may be indicative of data calculated by a function that enables processing of raw data generated by the detection means in advance. Thus, a logarithmic function for linearizing the raw data can be used.

It is apparent from the foregoing discussion that the present invention is not limited to the design of the sample dispensing system shown in FIG. 7; Some skilled in the art will be able to design the system for n greater than 6 using the information provided.

A detailed overview of the linear and nonlinear standard addition methodology is described in Frank et al. (Anal. Chem. Vol. 50, No. 9, August 78) and Saxberg et al. (Anal. CHem. Vol. 51, No. 7, June 1979). It is.

Preferred embodiments of the test specimen of the invention according to FIG. 3 include a catalyst compound but a calibration compound (each 6a)_One And 6'a_One) Does not include 1 detection part, catalyst compound and calibration compound of the first concentration (6a each)₂ And 6'a₂1 detection section, a catalyst compound and a calibration compound of a second concentration (6a each)₃ And 6'a₃1 detector and background absorbance (6c each) And one detector for 6'c). The latter detector, which does not contain a calibration compound or a catalyst compound, measures the background absorbance of the sample and allows this to be taken into account in the calibration process.

10 shows a pre-programmed verification method for calibration results and calibration data, whereby verification of the measurement results is confirmed by defining a "validation window" 29 for verification and calibration measurements. By this means, it is possible for the test object test system to limit all data to verified and useful concentration ranges, such as 30 to 600 mg / dL glucose, and the verification range of optical density, for example to 0.1 to 0.9. Similarly, the computing means makes the range verified by more generally constraining the slope and coefficient c ₀ to c _(n-1) , which is particularly useful in nonlinear polynomial equations. A verification measurement population located within the boundaries of verification window 29 is shown in FIG. 10; See descriptions 25b to 27b and 30.

Statistical evaluation and linear regression analysis can be used to verify the results more strongly. Since the quality of the calibration can be determined by the correlation coefficient r ² and the confidence interval, the specimen test system may not display the measurement results if the correlation coefficient is below a pre-programmed threshold. Alternatively, the calculating means may calculate the tolerance range or concentration range of the result based on the calculated confidence interval. By using these methods, a high degree of control of quality, which has been known and only available today using sophisticated and expensive laboratory methods and equipment, is also available to the average patient. More importantly for the patient / user, it is possible to guarantee quality at the time of measurement, especially in the hospital setting.

In another embodiment of the present invention additional additional security is possible; In this case, the test specimen system is operated to correlate the concentration of the inert dye with the amount of calibration compound used in the calibration process. The calibration formulation consists of a calibration compound and an inert dye, which are to be configured at a predetermined fixed ratio with each other before being applied on a predetermined detection portion of the test specimen. Thus, the computational means of the test object test system tracks slight fluctuations in the waste volume of the calibration compound when the detection means is operated to measure inert dye concentrations at wavelengths different from those for evaluating the reaction of the test object with the indicator compound, and Has the ability to correct

In addition, it becomes more reliable because it becomes possible to control the administration of the calibration formulation and the manufacture of the coating process. The inert dyes mentioned above are preferably brilliant blue G; Brilliant black BN; New coxin; Tarrazine; Indigo carmine; Sunset yellow, chamocin; Ponso 4R; Direct blue 2B; And / or water-soluble derivatives of malachite green.

By integrating the calibration process and the verification method, the specimen test system of the present invention not only has inherent interference such as different blood type and hematocrit concentrations, but also drugs or vitamin C which can affect the measurement results and modify the measurement results. It provides reliable results by offsetting exogenous interferences such as nutritional supplements. Since calibration of the specimen test system is parallel to the measurement, different environmental variables, such as temperature at the actual measurement, do not affect the accuracy of the measurement results.

In addition, production variables, such as variations in the thickness of the center layer, are compensated for by the integration of the calibration process and active ingredient aging, so that loss of enzyme activity can be traced and offset, for example, to extend the shelf life of the product.

11 illustrates various embodiments and shapes of the test strips of the present invention employed in various test test systems.

12 illustrates inserting an object test strip into an object test system. In a preferred embodiment, the specimen test strip is designed to have convex protrusions 10 on the side located on one major side of the test strip on which the sample application 9 is located.

As shown in FIG. 13, this structure facilitates the application of capillary blood samples from the arm or finger of the patient.

In another embodiment of the present invention as shown in FIG. 14, a plurality of specimen test members are disposed symmetrically around a center point to form the specimen test disc 31 with the sample application section 39 disposed to face the outer surface. do. This exemplary specimen test disc 31 according to FIG. 14A includes nine specimen test members of the present invention. As illustrated in FIG. 14B, the specimen test disc 31 is covered by a disc cover consisting of a top layer 32 and a bottom layer 33. The disc cover bottom layer 33 may be provided with a layer 34 which absorbs moisture. The top layer 32 and the bottom layer 33 of the disc cover have breakthroughs arranged integrally with each other, thereby forming the optical window 35 in which the specimen test member used in the current measuring method is located.

Adjacent to the optical window 35 on the outer end of the top layer 32 of the disc cover and the bottom layer 33 of the disc cover is a notch 36 for exposing the sample application part 39 of the measurement cell. Preferably, the test disc 31 may be provided with a fitting notch 38 which may be located at the inner edge of the disc 31. During the measurement process, only the specimen test member currently used for measurement of the specimen is exposed by the optical window as shown in FIG. 14C. The specimen test disc 31 can rotate around its center point, so that a new specimen test member can be brought to an appropriate position as needed.

By the test object disk means, it is possible to arrange a plurality of test object members in a relatively small area. As can be seen from the size comparison of the test strip and the test strip as shown in FIG. 15, placing the same number of test members on the test strip requires much more area and material. Nine specimen test members 41 enter the unit area 40 of the specimen test disc 31, while the same area 42 can only accommodate three specimen test strips. However, a reduction in test strip size is not possible, because smaller strips would be impractical as it would be difficult for the patient to manipulate.

16A and 16B show the specimen test disc included in the meter, in which the sample application sections 39, 43b further protrude from the meter housing.

In addition to the specimen test strip, in the case of the specimen test disc, the measurement device (the specimen test system) can be made left-handed and right-handed as shown in FIG. 17. If left-handed use is desired according to FIG. 17A, the specimen test strip 57 is inserted into the sample applicator 43a for receiving physiological body fluid protruding from the meter, the meter housing 58 from the bottom side. After the measurement is completed, the specimen concentration is shown on the specimen test system display 54. Likewise, the right-handed model according to FIG. 17B can be realized by rotating the display 54 of the specimen test system 59 by rotating the content shown on the display by 180 ^° , inserting the specimen test strip 57 from the top into the meter.

Fig. 18 illustrates the possibility of placing another test piece of space-saving test object. In this embodiment, the test object is continuously arranged side by side to form an ammunition form 44 where the sides protrude to form the sample application part 9. In this bullet zone, the area between the two specimen test members is provided with a perforation or dashed line 46 that separates the used specimen test member 45 from the specimen test cartridge 44 that has not yet been used. Fold or zigzag along the dashed line 46 to easily fill a small container, making it possible to create a specimen test device ammunition stack 48, which makes it much easier to dispense a single specimen test piece of specimen test ammunition. .

Specimen  Manufacturing method of test member

The specimen test member of the present invention, which is produced in the form of a disk or strip, can be easily manufactured by processes such as printing, dye punching, and lamination by those skilled in the art. The design of the specimen test member allows for a simple and cost effective manufacturing process that is desirable but not essential for continuous properties.

In the first step of the manufacturing method, a pattern of a sample distribution system is formed by generating a high surface energy portion and a low surface energy portion on a substrate. In a preferred embodiment, the high surface energy portion that forms the sample path and the detector on the first and second surfaces is created by applying a hydrophilic formulation on the hydrophobic surface of the substrate. As described above, it is also possible to make the high surface energy portion and the low surface energy portion by applying a hydrophobic "conductor member" pattern on the hydrophilic surface. If the substrate has intermediate hydrophobic properties, it is also possible to print the hydrophilic pathway in a surrounding hydrophobic pattern.

Substrates include glass, polyvinyl acetate, poly-methyl-methacrylate, poly-dimethyl-siloxane, polyester resins containing fluorene rings and polyesters, polycarbonates and polycarbonate-polystyrene graft copolymers, terminally modified poly Carbonate, polyolefin, cycloolefin and cycloolefin copolymers, and / or olefin-maleimide copolymers.

The application of a hydrophilic pattern on the hydrophobic substrate and / or the application of a hydrophobic "conductor member" on the hydrophilic substrate is preferably achieved using a flexographic- or lithographic printing process.

Flexography allows for high resolution printing on rotary presses and high speed production. This technique is an established technique for printing on polymer film substrates and is widely used in the packaging industry. The optical detection process shown in FIG. 8 requires low viscosity transparent clear ink for hydrophilic patterns. Low viscosity inks are preferred for obtaining thin and uniform coatings of about 2-4 microns. The optical window of the ink needs to fall within the wavelength range where the indicator dye absorbs light after the chemical reaction. Properties required for hydrophobic inks are, apart from hydrophobicity, less stringable and should be able to be used to decorate the test strip or disc with the desired color. Four-color flexographic-printing machines are operating in practice and do not cause any operational problems. This is the same for a lithographic device.

Even if a solvent base or UV curable ink can be used for the production of the test object to be tested, electron beam (EB) curable ink is much more preferable. These inks provide the best resistance to mechanical and chemical factors and contain 100% polymer with pigments depending on the pigment, but they contain no volatile organic solvents and photoinitiators, which have been shown to affect the stability of sensor chemistry. Do not. This positive result on performance characteristics is due to the ability of the electrons to form crosslinked polymer films and penetrate the surface.

Inks used for EB curing utilize the polymerization capacity of acrylic monomers and oligomers. Acrylic chemistry has special meaning in today's inks (6 J.T. Kunjappu. "The Emergence of Polyacrylate in Ink Chemistry," Ink World, February, 1999, p. 40). The structure of acrylic acid, the simplest acrylic compound, is as shown in the following formula (I).

The double bonds in the acrylic moiety open during the interaction (initiation) with the electrons to form free radicals, which act on other monomers to form chains to form (extended) polymer polymers. As described above, the polymerization induced by light irradiation does not require an initiator from the outside because the light irradiation itself generates free radicals, so that no starting species remains during the coating.

Various acrylic monomers can be used for EB curing and pre-polymers such as bisphenol A, epoxy acrylate and polyester / polyether acrylate from simple acrylates such as 2-phenoxyethyl acrylate and isooctyl acrylate (R. Golden. J. Coatings Technol., 69 (1997), p. 83). According to this curing technique, it is possible to design "functional inks" that focus on desirable physicochemical properties that are required for other inks and thus do not require solvents or curing systems that complicate the process.

Suitable hydrophobic inks contain monomers, oligomers, and prepolymers having hydrophobic functional groups such as isooctyl acrylate, dodecyl acrylate, styrene derivatives or systems with partially fluorinated carbon chains.

Inks having hydrophilic functional groups can be made by extensive selection from crosslinkable water soluble polymers, and useful are acrylics prepared from polyalcohols, polyethylene-glycols, polyethylene-oxides, vinylpyrrolidones, alkyl-phosphocholine derivatives and other materials. Late derivatives; Especially useful are organo-modified silicone acrylates, which are crosslinkable species of organo-modified polysiloxanes. Suitable coatings generally have a contact angle with water of less than 25 ^° and their surface energy generally exceeding 70 mN / m.

The second step of the method of preparation is an enzyme or other compound which performs a catalytic or non-catalytic reaction with the specimen on a predetermined detection portion of a sample distribution system formed on a substrate providing a first surface, and, if necessary, Applying a catalyst formulation comprising a coenzyme and an indicator dye and a calibration formulation containing different concentrations of the calibration compound or the specimen on a predetermined detection portion of the sample distribution system formed on the substrate providing the second surface. Application is carried out.

The accuracy of this deposition process is very important and also determines the accuracy and performance of the specimen test piece. Preferably both formulations are applied with the aid of a high precision ink jet system or a piezoelectric print head. Catalytic and calibration formulations should be made to dissolve well by physiological fluid samples. Preferably, they are preferably aqueous. Thus, these inks are usually made from water, enzymes, indicators, or calibration compounds, respectively, and are dried at slightly elevated temperatures. The main feature of these ink formulations is that they quickly reconstitute chemical components after sample application, without affecting the hydrophobic portion of the specimen test member.

The next step includes a lamination process, in which a base layer and a cover layer providing the first and second surfaces of the sample dispensing part are laminated on the center layer, thereby the first and second surfaces of the base layer and the cover layer. The distance between them is determined. The center layer provides a discontinuity in the portion where the sample distribution is formed on the first and second surfaces of the base layer and the cover layer, creating a cavity for the sample distribution. The high surface energy pattern and the low surface energy pattern formed on the first and second surfaces of the base layer and the cover layer must be aligned to coalesce with each other so that a functional sample distribution is formed between the first and second surfaces. do.

Accurate xy-registration of the base and cover layers is critical to the device's functionality, and if this consistency is not achieved, the sample distribution system will not function properly. The consistency tolerance for good performance is +/- 5% of the hydrophilic path width. Because of the relatively large discontinuities in the material relative to the size of the hydrophilic pathway, the application of the center layer, which may be a double-sided adhesive tape with a preferred thickness of 80 microns, is less difficult. This correspondence is especially important in continuous production lines where the substrate is advanced from a few meters to tens of meters of segmentation. The expansion of the substrate and the web tension make it more difficult to achieve consistency in the x-direction (web travel direction) than in the y-direction perpendicular to the web travel direction.

A method of making an elastic polymer film of the present invention, which provides accurate pattern matching of a first surface and a second surface, is shown in FIG. 19, which shows part of a continuous web manufacturing process. In the first production step according to FIG. 19A, the pattern of the sample dispensing system 6 of the base layer and the cover layer is printed on one web substrate 49 representing the specimen test member and the strip material, respectively. As shown in FIG. 19, the printed pattern of the sample dispensing system 6 is disposed on the web substrate 49 in such a way that the two sample dispensing systems are opposed to each other and later connected to each other at the portion forming the sample application. Therefore, the positions of the predetermined detectors 6a and 6'a are fixed relative to each other and thus remain unaffected by the expansion of the material or the web tension.

Dotted line 50 represents a cut line that later separates the test strip, and dashed line 51 represents a line for later folding of the web substrate.

After printing the sample distributor, the detectors 6a, 6'a of the sample distribution system are coated with a catalyst formulation and a calibration formulation. For example, the detection section 6'a in the lower row of web substrate 49, which will represent the first surface of the specimen test member, will be coated with a catalyst formulation containing enzyme and indicator, while the second surface of the specimen test member will appear. Detection 6a in the upper row of web substrate 49 is coated with calibration formulations containing calibration compounds of varying degrees of knol; One calibration formulation here (eg, located at 6a ₁ ) contains no calibration compound and delivers a reading of physiological fluid at the detection stage.

Thereafter, an additional layer is deposited on either of the surfaces (eg, surface 2a of base layer 2) to form the center layer 52 of the specimen test member, as shown in FIG. 19B. The center layer 52 can be made of double-sided adhesive tape and provides a breakthrough 5 that exposes the sample dispensing system 6, thereby forming a cavity for the sample dispensing system in the specimen test member after the final assembly step.

Subsequently, as shown in FIG. 19C, for example, using a folding iron, the test object of the present invention is folded into two sides along the fold line 51 to form a folded web 53 as shown in FIG. 19D. . The press roller can then be used to make the connection between the center layer, the base layer and the cover layer more rigid.

Finally, the laminated web 53 is cut and perforated to the desired product shape, with dashed line 50 showing an exemplary shape of the final specimen test strip on the web 53 prior to the separation step. In the manufacturing method shown in FIG. 19, the sample dispensing system is easier in comparison to the single sheet method, since the upper part of the substrate can be folded over the bottom part without the risk of losing consistency in the x-direction of the web. It is possible to achieve the exact correspondence of the first and second surfaces forming a.

The present invention provides calibration and quality control in dry reagent test strip formats without the need for users to intervene in calibration and quality control processes with precise control of strip performance in sample analysis without requiring excessive strip fabrication processes. Provide an object test system incorporating the means.

1 is a perspective view of one embodiment of an object test member of the present invention provided in the form of a test strip.

FIG. 2 is a perspective view of the embodiment according to FIG. 1, showing an enlarged view of a sample dispensing system. FIG.

3 is a schematic perspective view of the device according to FIG. 1, showing three layers individually.

4 shows various different discontinuities in the center layer forming the sample cavity with the first and second surfaces.

5 is a cross-sectional view of a detection portion of a sample dispensing system constructed by a hydrophobic guiding member.

6 is a cross-sectional view of another embodiment of a detector of a sample distribution system using a hydrophilic pathway.

7 illustrates various embodiments of a sample distribution system with various different detectors and path patterns suitable for various different calibrations.

8 is a cross-sectional view of the sample dispensing system of FIG. 6 in conjunction with a light emitter and detection means.

9 is a graph showing a method of calculating a sample specimen concentration using a standard addition method.

10 is a graph illustrating a verification method of calculation results and calibration data.

11 illustrates specimen test strips of different shapes.

12 shows an example of application of a test strip of the invention with a meter.

FIG. 13 illustrates an object test system having an embedded object test strip. FIG.

It is a figure which shows the structure of the test object test disc.

15 is a diagram illustrating a comparison between an object test strip and an object test disk.

FIG. 16 illustrates an object test system equipped with an integrated object test disc.

FIG. 17 illustrates an object test system equipped with an object test strip of left-handed type and right-handed type.

FIG. 18 is a stack of specimen test ammunition bands and ammunition stacks stacked together.

It is a figure which shows the manufacturing process of the test object with a strip shape. The layers shown in FIGS. 5, 6 and 8 are not drawn to scale, in particular the thicknesses of layers 16, 17, 18 and 19 are exaggerated.

Claims (35)

A specimen test member for measuring the concentration of at least one specimen in a physiological fluid sample having a first surface 2a and a second surface 4a spaced apart from each other by a predetermined distance, both of which surfaces Is provided with two substantially uniform patterns forming a high surface energy portion and a low surface energy portion, wherein the high surface energy portion and the low surface energy portion are aligned so as to merge well with each other and have two or more detection portions 6a. A test member for forming a sample dispensing system (6), wherein the applied physiological fluid is limited to a high surface energy portion. The center layer (3) according to claim 1, wherein the distance between the first surface and the second surface is disposed between the base layer (2) having the first surface and the second surfaces (2a, 4a) and the cover layer (4). The test object to be tested is determined by. 3. The center layer 3 has a discontinuity 5 which forms a hollow cavity with the base layer and the first and second surfaces 2a, 4a of the cover layers 2, 4. The hollow cavity test piece is larger in size than the sample dispensing portion (6) formed by the high surface energy portion on the first and second surfaces (2a, 4a). The test according to any one of the preceding claims, wherein the high surface energy portion is made by applying a crosslinkable and / or insoluble hydrophilic and / or amphiphilic reagent on the first and second surfaces 2a, 4a. Absence of water test. The method of claim 4, wherein the hydrophilic reagent is selected from polyols, polyethylene-glycols, polyethylene-oxides, vinylpyrrolidone and organo-modified polysiloxanes or functionalized derivatives derived from alkyl-phosphocholine polyethylene-glycol copolymers. The test piece to be tested. The method according to any one of the preceding claims, wherein the first surface 2a and the second surface 4a are hydrophobic and not wetted by physiological fluids, and in particular the UV, near-infrared and / or visible portions of light, in particular electromagnetic spectrum The test member to be transmitted through light. The low surface energy portion is made by applying a hydrophobic composition on the first and second surfaces 2a, 4a, wherein the hydrophobic composition is moistened with physiological fluid. The test piece to prevent from falling. 8. The test piece of claim 7, wherein the hydrophobic composition contains a system having isooctyl acrylate, dodecyl acrylate, styrene derivative or partially fluorinated carbon chain. 8. The surface of claim 7 wherein the first surface 2a and the second surface 4a are hydrophilic and can be wetted by physiological fluids, which transmit light, in particular UV, near infrared and / or visible light of the electromagnetic spectrum. The test piece to be tested. The test piece according to claim 9, wherein the first surface (2a) and the second surface (4a) are made hydrophilic by physical or chemical vapor deposition of a hydrophilic compound. The base layer 2 and the cover layer 4, which provide the first and second surfaces 2a, 4a, are glass, polyvinyl acetate, poly-methyl-methacrylate according to any one of the preceding claims. , Poly-dimethyl-siloxanes, polyester and polyester resins containing fluorene rings, polycarbonates and polycarbonate-polystyrene graft copolymers, terminally modified polycarbonates, polyolefins, cycloolefins and cycloolefin copolymers, and / or A test piece to be made of a material selected from olefin-maleimide copolymers. The method according to any one of the preceding claims, The n predetermined detection portions 6'a of the first surface 2a are coated with a catalyst formulation for promoting the detection of the specimen in physiological body fluids, Wherein the n predetermined detection portion of the second surface (4a) (6a) is the calibration compound of the n different in the m blank formulations and concentration-for the n calibration consisting of m Formulations Formulations (where n is greater than 2 Integer, m is an integer of 1 or more, where n > m ) Test piece to be coated with. 13. The specimen test member according to claim 12, further comprising a detection section (6c) which does not contain a catalyst compound and contains no calibration compound to enable measurement of a background signal. 14. The test according to claim 12 or 13, wherein the catalyst formulation coated on the n predetermined detection portions 6'a of the first surface 2a is contained in a physiological bodily fluid sample using transmission or absorption photometry. No test specimen to allow measurement of water concentration. 13. The calibration compound according to claim 12, wherein the calibration compound contained in the n - m predetermined detection sections 6a of the second surface 4a is the same as or substantially equivalent to the test specimen, so as to produce the same chemical reaction as the test specimen in the physiological bodily fluid sample. The test piece to be tested can be derived from the catalyst formulation. The test member of claim 15, wherein the calibration compound is glucose. The test article of claim 12, wherein the catalyst formulation contains as a reactive component a promoter that performs a catalytic or non-catalytic reaction with the test article, and / or a coenzyme, and an indicator that generates an optically detectable product. No test. 18. The test member of claim 17, wherein the promoter is an enzyme selected from dehydrogenase, kinade, oxidase, phosphatase, reductase and / or transferase. The test member of claim 18, wherein the promoter is an enzyme specific for glucose. 18. The test according to claim 17, wherein the indicator for determining the test concentration is selected from aromatic amines, aromatic alcohols, azines, benzidines, hydrazones, aminoantipyrines, conjugated amines, conjugated alcohols, and / or aromatic and aliphatic aldehydes. Absence of water test. 21. The calibration formulation according to any one of claims 12 to 20, wherein the calibration formulation applied to the predetermined detection section 6a of the second surface 4a is applied to the inert water-insoluble dye at a predetermined fixed ratio with respect to the calibration compound. Containing, allowing the appropriate reading device to evaluate the concentration of the calibration compound in the calibration formulation at a wavelength different from the wavelength used to determine the reaction product between the catalyst formulation and the test article. The method of claim 21, wherein the inert water-insoluble dye comprises Brilliant Blue G; Brilliant black BN; New coxin; Tarrazine; Indigo carmine; Sunset yellow, chamocin; Ponso 4R; Direct blue 2B; And / or a water soluble derivative of malachite green. The test piece according to any one of the preceding claims, wherein the sample application part (9) is located at the distal end of the convex side protrusion (10) on one side of the test piece. An object test arrangement structure comprising a plurality of devices according to any one of the preceding claims, wherein the devices are arranged symmetrically around a center point, such that the sample application section 39 is aligned outward. The test piece test batch structure which forms (31). An object test arrangement structure comprising a plurality of devices according to any one of the preceding claims, wherein the devices are arranged in a linear manner so that the specimen test ammunition 44 with the sample application section 9 protruding to the side. The test piece test arrangement structure, which is to be formed, and the application portion 39 is disposed to face outwardly. A process of generating a high surface energy portion and a low surface energy portion on a base layer 2 having a first surface 2a, wherein n high surface energy portions (where n is an integer greater than 2) of a predetermined detection portion ( Forming hydrophilic pathways with 6'a), Applying a corresponding pattern of the high surface energy portion and the low surface energy portion to the cover layer 4 having the second surface 4a, Coating a catalyst formulation on the n detection portions 6'a of the first surface 2a, wherein the catalyst formulation detects the concentration of the specimen contained in the physiological bodily fluid sample using transmission or absorption photometry. To promote the process, 2 to the step of coating the n calibration formulations on n detection section (6a) of the surface (4a), said n calibration formulations is of m blank formulations and different calibration compound concentration n that - m of Formulations Wherein m is an integer greater than or equal to 1 and n > m , wherein the calibration compound is the same or substantially the same as the test specimen so that the same chemical reaction as the test specimen in the physiological bodily fluid sample can be induced in the catalyst formulation. Process that is, A first at an opposite portion of the center layer 3 having discontinuities 5 providing cavities to the sample dispensing system 6 formed by the high surface energy portions on the first and second surfaces of the base and cover layers; Laminating the layers of the surface and the second surface, Process of punching or cutting the laminated sheets into final shape Method for producing a test object to be tested, comprising: 27. The method of claim 26, wherein the high surface energy portion is made by applying a crosslinkable and / or water-insoluble hydrophilic and / or amphiphilic reagent to the first and second surfaces. 28. The method of claim 27, wherein the high surface energy portion is printed on the first and second surfaces by flexographic or lithographic means. 27. The test of claim 26, wherein the low surface energy portion is made by applying a hydrophobic compound on the first and second surfaces, wherein the hydrophobic compound prevents the coated portion from being wetted by physiological fluids. Method of manufacturing the member. 30. The method of claim 29, wherein the first and second surfaces are made hydrophilic by physical or chemical vapor deposition of a hydrophilic compound. 31. The test piece of any of claims 26 to 30, wherein the high surface energy portions of the first and second surfaces are physically located higher than the low surface energy portions by etching or embossing. Manufacturing method. 32. The method of claim 29, wherein the low surface energy portion is printed on the first surface and the second surface by flexography or lithography. 33. The base layer (2) and the cover layer (4) according to any one of claims 26 to 32 are formed on one elastic substrate (49) at a fold line (51) located at the center in the longitudinal direction. Folded along, the center layer (in such a manner that the sample dispensing system 6 having the predetermined detection portions 6'a, 6a of the first surface 2a and the second surface 4a are aligned and aligned so as to integrate well). The manufacturing method of the test object to be sealed which has 3). The n predetermined detection portions 6'a of the first surface 2a are coated with a catalyst formulation for promoting the detection of the specimen in physiological body fluids, The 2 n of the predetermined detection of surface (4a) (6a) is of m blank formulations and, n containing a calibration compound having different concentrations - n of calibration formulations made of m formulations (where n is 2 A test member or a test test batch according to any one of claims 1 to 25, coated with a greater integer, m being an integer greater than or equal to n > m ), 2 n of the predetermined position within the detection unit, and that obtain the n results from 2 n predetermined detection part of the detection means for detecting a change in absorbance of the physiological sample, and Equation

Calculation means for calculating one regression coefficient for verifying the quality of the n - m correction coefficient of the polynomial correction equation and the calculated n - m correction coefficient of the correction equation An object test system for measuring the concentration of the object in the physiological sample solution comprising a. A method for measuring the concentration of one or more specimens in a physiological sample, A process of applying a physiological sample to an object test member having a first surface 2a and a second surface 4a spaced apart from each other by a predetermined distance, wherein both surfaces have a high surface energy portion and a low surface area. Two substantially uniform patterns are provided which form a surface energy section, wherein the high surface energy section and the low surface energy section are arranged to integrate well with each other to provide a sample distribution system 6 having two or more detection sections 6a. Process, wherein the applied physiological fluid is limited to a high surface energy portion, Detecting signals generated by various different detection units, and The process of correlating signals to measure the amount of specimen (s) in a physiological sample Method for measuring the concentration of one or more specimens of the physiological body fluid sample comprising a.

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