JP2005300528A - Analysis element used for inspection method of specimen - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an analysis element used for a blood inspection method that can be operated easily and is capable of measurement rapidly. <P>SOLUTION: The multiple item measurement dry analysis element is used for a specimen inspection method for obtaining measurement results from information on 1,000 pixels or higher for one item and measuring a plurality of items simultaneously, by using an area sensor as a detector. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to an analytical element used in a blood test method for humans and other animals. Analytical elements used in testing methods that use human and animal body fluids, urine, fresh water, seawater and soil extracts, agricultural products, marine products and processed food extracts, and liquids used in natural science research as specimens About.

  Conventionally, a method for diagnosing a human disease using blood, urine, or the like as a specimen has long been performed as a method that can be easily diagnosed without damaging the human body.

In particular, blood can be diagnosed for many test items.
As an analysis method for such multi-item inspection, a wet chemistry analysis method has been conventionally developed. This is a method using a so-called solution reagent. An apparatus using a wet chemistry analysis method for multi-item inspection is generally complicated because it combines a large number of reagent solutions corresponding to multiple items and their handling, and the handling and procedure of the apparatus is simple. Absent.

On the other hand, a method that can easily perform the analysis is being sought.
As one method, a so-called dry chemistry analysis method has been developed that does not use a solution for analysis, that is, contains reagents necessary for detection of a specific component in a dry state (Non-patent Document 1). .

However, in both wet chemistry and dry chemistry, when the sample is blood, whole blood is usually not used, and after removing blood cell components, it is subjected to analysis in the form of plasma or serum. As a method for removing blood cell components, conventionally, blood cell separation is performed by a method using centrifugal force, which requires a centrifugal separation operation and has a problem that it takes a long time until detection. On the other hand, a device for separating blood cells by a method using a filter has been developed (Patent Document 1), and the time required for blood cell separation has been shortened. However, separation of blood cells and detection are separate operations, and the time can be sufficiently shortened. I can't say that.
In response to this, there has been proposed an apparatus capable of analyzing many items without using a blood cell separation operation by using a dry chemistry analysis method and combining with a centrifugal separator (Patent Documents 2 and 3). However, the operation of the centrifugal separator is required, and it has not yet been sufficient to satisfy the convenience. There is also a problem that the time until detection is long.

On the other hand, in an aging society, blood tests that can easily measure the health status are becoming increasingly important, and even in lifestyle-related diseases, it is a means by which changes in disease status can be easily known. . In elderly people and lifestyle-related diseases, it is necessary to observe the state of health / disease over time, and more and more scenes require blood tests. Therefore, there is a demand for a method that allows not only medical personnel but also a patient to collect blood by himself and analyze it easily and quickly. Furthermore, nosocomial infections have become a major social problem in recent years, and in particular, protection from infection from blood is desired.
On the other hand, an analyzer that integrates means from blood collection to analysis by combining blood collection with a needle, blood cell separation by filtration and centrifugation, and wet chemistry analysis by an electrode method has been proposed (Patent Literature). However, 4) has not yet been fully satisfactory in terms of simplicity of operation. In addition, variations in measured values may occur, and it has not been satisfactory from the viewpoint of measurement accuracy in clinical examinations.
Furthermore, in the medical field, it is required to perform from sample collection to analysis and detection more quickly. An analyzer that integrates means from blood collection to analysis and detection by combining with a photodetector has been proposed (Patent Document 5).
JP 2000-180444 A Special table 2001-512826 gazette JP-T-2002-514755 JP 2001-258868 A JP 2003-287533 A Yuzo Iwata, “11. Other Analytical Methods (1) Dry Chemistry”, Clinical Chemistry Practice Manual, Medical School, 1993, Examination and Technology Extra Number, Vol. 21, No. 5, p. 328-333

As described above, a method for examining a specimen with respect to multiple items is required to be simpler with better operability. Moreover, when used for clinical examination, it is necessary to be safe and to have sufficient measurement accuracy. In addition, there is a need for an inspection method that can perform detection more quickly for more items than before.
An object of the present invention is to provide an analytical element used in a blood test method that is simple in operation and can be rapidly detected.
Another object of the present invention is to provide an analytical element used in a blood test method that can be rapidly detected for many items and that is safe and has sufficient measurement accuracy. .
A further object of the present invention is a test method that uses human or animal body fluids, urine, fresh water, seawater and soil extracts, agricultural products, marine products and processed food extracts, and liquids used in natural science research as specimens. It is to provide the analytical elements used.

As a result of intensive studies, the present inventors have found that the above object can be achieved by using a multi-item measurement dry analysis element and a specific detector in combination under specific conditions.
That is, the present invention achieves the above object by the following configuration.
1. A multi-item measurement dry analysis element for use in a specimen testing method that uses an area sensor as a detector to obtain a measurement result from information of 1000 pixels or more per item and can simultaneously measure a plurality of items.
2. The multi-item measurement dry analysis element has a flow path, a color reaction reagent, and a part carrying the color reaction reagent,
At least one of the width, depth, and length of the flow path is 1 mm or more,
And the width of the part carrying the color reaction reagent is at least twice the width of the flow path and / or the length of the part carrying the color reaction reagent is 0.4 times the length of the flow path The above 1. Multi-item measurement dry analysis element described in 1.
3. 2. A filtration part containing a water-insoluble substance having an equivalent circle diameter of 5 μm or less and a length equal to or longer than the equivalent circle radius. Multi-item measurement dry analysis element described in 1.
4). 2. A filtration part containing fibers having an equivalent circle diameter of 5 μm or less. Multi-item measurement dry analysis element described in 1.
5). 2. A filtration part containing a fiber having a circle equivalent diameter of 5 μm or less and a porous membrane. Multi-item measurement dry analysis element described in 1.
6). 2. A filtration part containing a glass fiber having an equivalent circle diameter of 5 μm or less and a porous membrane. Multi-item measurement dry analysis element described in 1.
7). 7. The multi-item measurement dry analytical element according to any one of 2 to 6, wherein the dry multilayer film is used as a reagent layer in a portion carrying the color developing reagent.
8). 2. The dry multilayer film formed by adhering a porous membrane is used as a reagent layer in the portion carrying the developer reagent. Or 3. Multi-item measurement dry analysis element described in 1.
9. 2. The dry multilayer film to which fine particles having a diameter of 100 μm or less are adhered is used as a reagent layer in the portion carrying the color developing reagent. Or 3. Multi-item measurement dry analysis element described in 1.
10. 2. The portion carrying the color reaction reagent is a cell connected to the flow path. Or 3. Multi-item measurement dry analysis element described in 1.
11. 2. The specimen is supplied to the reagent through the polymer porous body, and the dry multilayer film is used as the reagent layer in the portion carrying the developer reagent. Or 3. Multi-item measurement dry analysis element described in 1.
12 2. The specimen is supplied to the reagent through the space marked in the flow path, and the dry multilayer film is used as the reagent layer in the portion carrying the color reaction reagent. Or 3. Multi-item measurement dry analysis element described in 1.

13. In a multi-item measurement dry analytical element for use in a specimen testing method capable of measuring multiple items at the same time using a line sensor as a detector,
The multi-item measurement dry analysis element has a flow path, a color reaction reagent, and a part carrying the color reaction reagent,
At least one of the width, depth, and length of the flow path is 1 mm or more,
And the width of the part carrying the color reaction reagent is at least twice the width of the flow path and / or the length of the part carrying the color reaction reagent is at least 0.4 times the length of the flow path And
A multi-item measurement dry analytical element having a filtration part containing a water-insoluble substance having an equivalent circle diameter of 5 μm or less and a length equal to or longer than the equivalent circle radius.
14 In a multi-item measurement dry analytical element for use in an analyte test method capable of measuring multiple items simultaneously using an electrochemical detector as a detector,
The multi-item measurement dry analytical element has a flow path, a reaction reagent, and a part carrying the reaction reagent,
At least one of the width, depth, and length of the flow path is 1 mm or more,
A multi-item measurement dry analytical element having a filtration part containing a water-insoluble substance having an equivalent circle diameter of 5 μm or less and a length equal to or longer than the equivalent circle radius.
15. 1. a blood collection device having two or more parts slidable with respect to each other; A blood collection unit comprising: the multi-item measurement dry analysis element as described above; and loading the element into the instrument, and defining a sealed space inside the apparatus so that the pressure can be reduced by sliding while maintaining a substantially airtight state.
16. 15. The blood collection instrument described above, which has a puncture needle having a diameter of 100 μm or less and a tip angle of 20 degrees or less. The blood collection unit described in 1.
17. 12. A blood collection device having two or more parts slidable with respect to each other; A blood collection unit comprising: the multi-item measurement dry analysis element as described above; and loading the element into the instrument, and defining a sealed space inside the apparatus so that the pressure can be reduced by sliding while maintaining a substantially airtight state.
18. 16. The blood collection instrument described above, wherein the blood collection instrument has a puncture needle having a diameter of 100 μm or less and a tip angle of 20 degrees or less. The blood collection unit described in 1.
19. 2. A liquid for testing environment-related substances is used as a specimen. Multi-item measurement dry analysis element described in 1.
20. 2. A liquid for agriculture, fisheries or food inspection is used as the specimen. Multi-item measurement dry analysis element described in 1.
21. 2. A liquid used for natural science research is used as a specimen, Multi-item measurement dry analysis element described in 1.

In short, by making it possible to detect multiple items at the same time by any one of the following configurations (A) to (C), it is possible to quickly and more easily test the multiple items of the specimen.
(A) A multi-item measurement dry analysis element for use in a specimen inspection method capable of obtaining a measurement result from information of 1000 pixels or more per item using an area sensor as a detector and simultaneously measuring a plurality of items.
(B) In a multi-item measurement dry analytical element for use in a specimen testing method capable of measuring a plurality of items simultaneously using a line sensor as a detector,
The multi-item measurement dry analysis element has a flow path, a color reaction reagent, and a part carrying the color reaction reagent,
At least one of the width, depth, and length of the flow path is 1 mm or more,
And the width of the part carrying the color reaction reagent is at least twice the width of the flow path and / or the length of the part carrying the color reaction reagent is at least 0.4 times the length of the flow path And
A multi-item measurement dry analytical element having a filtration part containing a water-insoluble substance having an equivalent circle diameter of 5 μm or less and a length equal to or longer than the equivalent circle radius.
(C) In a multi-item measurement dry analytical element for use in a specimen testing method capable of measuring a plurality of items simultaneously using an electrochemical detector as a detector,
The multi-item measurement dry analytical element has a flow path, a reaction reagent, and a part carrying the reaction reagent,
At least one of the width, depth, and length of the flow path is 1 mm or more,
A multi-item measurement dry analytical element having a filtration part containing a water-insoluble substance having an equivalent circle diameter of 5 μm or less and a length equal to or longer than the equivalent circle radius.
In addition to the achievement of the above-mentioned objectives, it has been found that these structures do not leak red blood cells even when the amount of collected whole blood increases, and a sufficient amount of plasma can be supplied to the reagent. did. Furthermore, it has also been found that it is easy to perform a multistage reaction between a specimen and a reagent in stages.

ADVANTAGE OF THE INVENTION According to this invention, the analysis element used for the blood test method which is easy to operate and can perform a measurement rapidly is provided. In addition, an analytical element used for a blood test method that can perform measurement quickly for many items and that is safe and has sufficient measurement accuracy is provided.
Furthermore, according to the present invention, human or animal body fluids, urine, fresh water, seawater and soil extracts, agricultural products, marine products and processed food extracts, and liquids used in natural science research are used as specimens. Analytical elements used in the method are provided.

Hereinafter, the multi-item measurement dry analysis element of the present invention will be described.
The multi-item measurement dry analytical element of the present invention uses an area sensor, a line sensor or an electrochemical detector as a detector. First, the detector will be described.
[Detector]
Any area sensor may be used as long as it is arranged so that it can detect two-dimensional information by detecting light such as ultraviolet light, visible light, infrared light, or electromagnetic waves. it can. For example, CCD, MOS, photographic film, etc. are mentioned. Among these, CCD is preferable. By detecting a multi-item measurement dry analytical element using an area sensor, a measurement result can be obtained from information of 1000 pixels or more for one item, and a plurality of items can be measured simultaneously.
Any line sensor can be used as long as it is arranged so that it can detect one-dimensional information by detecting light such as ultraviolet light, visible light, infrared light, or electromagnetic waves. it can. For example, a photodiode array (PDA), a photographic film arranged so as to detect light in a slit shape, and the like can be mentioned. Among these, a photodiode array is preferable. By detecting the specific multi-item measurement dry analysis element using a line sensor, it is possible to measure a plurality of items at the same time.
Any electrochemical detector can be used as long as it can measure the amount of current, potential difference, electrical conductivity, and resistance in the conductive material medium. For example, a conductive electrode alone electrode such as a gold electrode, a platinum electrode, a silver electrode, or a carbon electrode, a silver / silver chloride electrode, an oxygen electrode, a composite electrode such as a modified electrode coated with an enzyme such as glucose oxidase, or a combination thereof. It is done. Among these, a modified electrode coated with an enzyme such as glucose oxidase is preferable. By detecting the above-described specific multi-item measurement dry analytical element using an electrochemical detector, it is possible to measure a plurality of items simultaneously.

Next, the multi-item measurement dry analysis element will be described in detail. Hereinafter, the case where an area sensor is used as a detector will be described. When a line sensor is used as a detector and when an electrochemical detector is used, it is applied in the same manner as in the case of an area sensor as long as the configurations described in (b) to (c) are satisfied.
[Multi-item dry analysis element]
The multi-item measurement dry analysis element has a flow path, a color reaction reagent, and a portion carrying the color reaction reagent, and at least one of the width, depth, and length of the flow path is 1 mm or more and the width of the part carrying the color reaction reagent is at least twice the width of the flow path and / or the length of the part carrying the color reaction reagent is the length of the flow path It is preferable that it is 0.4 times or more.
First, the flow path will be described.
"Flow path"
As described above, at least one of the width, depth, and length of the flow path is preferably 1 mm or more. More preferably, it is the range of 1 mm-100 mm, and the most preferable range is 1 mm-30 mm. Within this range, it is preferable because the sample can efficiently travel through the flow path.
The flow path can take any form as long as the specimen can pass through. Further, the flow path may be only one or may be branched into two or more. Moreover, although it can take any form, such as linear form and curvilinear form, it is preferable that it is linear form.

Any material can be used for the flow path as long as the passage of the sample proceeds efficiently. Specific examples include resins such as rubber and plastic, and silicon-containing substances.
Examples of the plastic or rubber include polymethyl methacrylate (PMMA), polycyclic olefin (PCO), polycarbonate (PC), polystyrene (PS), polyethylene (PE), polyethylene terephthalate (PET), polypropylene (PP), poly Examples include dimethylsiloxane (PDMS), natural rubber, synthetic rubber, and derivatives thereof.
Examples of the silicon-containing substance include amorphous silicon such as glass, quartz, and silicon wafer, and silicone such as polymethylsiloxane.
Among these, PMMA, PCO, PS, PC, glass, and silicon wafer are preferable.

The channel can be created on a solid substrate by a microfabrication technique. Examples of materials used include metal, silicon, Teflon (registered trademark), glass, ceramics or plastic, and rubber.
Examples of the plastic include PCO, PS, PC, PMMA, PE, PET, PP, and the like. Examples of rubber include natural rubber, synthetic rubber, silicon rubber, PDMS and the like.
Examples of the silicon-containing substance include amorphous silicon such as glass, quartz, and silicon wafer, and silicone such as polymethylsiloxane.
Particularly preferable examples include PMMA, PCO, PS, PC, PET, PDMS, glass, and silicon wafer.

Microfabrication technology for creating flow paths is, for example, microreactors-New generation synthesis technology-(published by CMC in 2003: Junichi Yoshida, Professor, Graduate School of Engineering, Kyoto University), Microfabrication Technology Applications-Photonics Electronics -Application to mechatronics-(2003, published by NTS edited by the Society of Polymer Science, edited by the Society of Polymer Science)
Typical methods include LIGA technology using X-ray lithography, high aspect ratio photolithography method using EPON SU-8, micro electric discharge machining method (μ-EDM), and high aspect ratio silicon processing method using Deep RIE. , Hot Emboss processing method, stereolithography method, laser processing method, ion beam processing method, and mechanical micro cutting method using a micro tool made of a hard material such as diamond. These techniques may be used alone or in combination. Preferred microfabrication techniques are LIGA technology using X-ray lithography, high aspect ratio photolithography using EPON SU-8, micro electrical discharge machining (μ-EDM), and mechanical micromachining.

  The flow path in the present invention can also be created by using a pattern formed using a photoresist on a silicon wafer as a mold, and pouring a resin into the pattern to solidify it (molding method). In the molding method, a silicon resin typified by PDMS or a derivative thereof can be used.

  It is desirable that the surface of the channel is treated or modified as necessary so that the specimen, particularly whole blood or plasma, can pass through quickly. The method of surface treatment and modification varies depending on the material constituting the flow path, but an existing method can be used. For example, plasma treatment, glow treatment, corona treatment, or a method using a surface treatment agent such as a silane coupling agent, polyhydroxyethyl methacrylate (PHEMA), polymethoxyethyl acrylate (PMEA), or acrylic polymer is used. And a surface treatment method.

The flow path may be a part of or a part of the multi-item measurement dry analysis element. In other words, the flow path can be created as a part of the multi-item measurement dry analysis element or by itself using a so-called microreactor or a microfabrication technique generally used for the microanalysis element.
For example, a method described in “Microreactor” (supervised by Junichi Yoshida, published by CMC) can be used as a method for creating a microreactor or a microanalytical element.

Next, the color developing reagent will be described.
"Development reagent"
The developer reagent is necessary for qualitative / quantitative analysis of the sample component to be measured, and reacts with the sample component to be colored, or develops color, fluorescence, luminescence, etc. Says something that emits light by action. In the present invention, it can be appropriately selected according to the type of specimen and the item to be measured. As an example, Fuji Dry Chem mount slide GLU-P (measurement wavelength: 505 nm, measurement component: glucose) and TBIL-P (measurement wavelength: 540 nm, measurement component: total bilirubin) manufactured by Fuji Photo Film Co., Ltd. may be mentioned. In the present invention, a dry reagent is used as the color reaction reagent included in the multi-item measurement dry analysis element. The dry reagent is a reagent used for so-called dry chemistry. Any reagent that can be used for dry chemistry can be used. Specifically, for example, Fuji Film Research Report No. 40 (Fuji Photo Film Co., Ltd., published in 1995) p. 83, clinical pathology, extra edition, special feature No. 106, new development of dry chemistry / simplified examination (Clinical Pathology Publication, published in 1997), and the like.

  When an electrochemical detector is used as the detector, for example, glucose oxidase (GOD), 1,1′-dimethylferrocene, and a carbon paste made of a mixture of graphite powder and paraffin are mixed instead of the color developing reagent. Using the enzyme electrode prepared by solidification as a working electrode, using a silver / silver chloride electrode as a reference electrode and a platinum wire as a counter electrode, it is possible to measure the current value that increases with the glucose concentration in the sample. More specifically, for example, Okuda, Mizutani, Yabuki et al. 290, 173-177 (1991) and the like.

Next, the portion carrying the developer reagent will be described.
When an electrochemical detector is used as the detector, it is the same as the portion of the area sensor carrying the color reaction reagent, except that the portion carrying the reaction reagent is carried by the reaction reagent. is there.
"Part carrying the color developing reagent"
As described above, the portion carrying the color reaction reagent is at least twice the width of the channel and / or the length of the portion carrying the color reaction reagent is the length of the channel. It is preferable that it is 0.4 times or more.
There may be only one part or two or more parts carrying the developer reagent. In the case of two or more, they may be collected in one place or arranged separately.
The portion carrying the color reaction reagent may be connected to the flow path, or the portion carrying the color reaction reagent may be incorporated in the flow path. Further, in the case of a form connected to the flow path, the portion carrying the color reaction reagent may be a cell. The cell may have any form as long as the width and / or length with respect to the flow path satisfy the above. Examples of the material of the cell include the same material as described above for the flow path. Moreover, a preferable thing is also the same.
A joining technique can be used for connection between the flow path and the portion carrying the developer reagent. The usual bonding techniques are roughly divided into solid phase bonding and liquid phase bonding, and the commonly used bonding methods are pressure bonding and diffusion bonding as solid phase bonding, welding, eutectic bonding, soldering as liquid phase bonding, Adhesion and the like are typical joining methods.
Furthermore, it is desirable to have a highly precise bonding method that maintains the dimensional accuracy without destroying microstructures such as flow path due to material deterioration or large deformation due to high temperature heating, such as silicon direct bonding, anodic bonding, surface Examples include activated bonding, direct bonding using hydrogen bonding, bonding using an HF aqueous solution, Au—Si eutectic bonding, and void-free bonding.
Further, bonding using ultrasonic waves, laser, etc., bonding using an adhesive, an adhesive tape, or the like may be used, or bonding may be performed only by pressure.

The portion carrying the color reaction reagent may carry the reagent in any form as long as it can carry the color reaction reagent. For example, a test paper, a disposable electrode, a magnetic material, a film for analysis and the like can be mentioned. In the case of a film, it may be a single layer or a multilayer.
Preferably, a dry multilayer film is used as a reagent layer in a portion carrying a color developing reagent. The dry multilayer film is preferable because all or a part of the reagents necessary for qualitative / quantitative analysis of the component to be measured in the specimen can be incorporated into one or more layers. Examples of the dry multilayer film include those used in the above-mentioned dry chemistry. Specifically, Fuji Film Research Report, No. 40 (Fuji Photo Film Co., Ltd., published in 1995) p. 83, clinical pathology, extra edition, special feature No. 106, new development of dry chemistry / simplified examination (Clinical Pathology Publication, published in 1997), and the like. It is preferable to use a dry multilayer film as a reagent layer in a portion carrying a color developing reagent because it is easy to perform a multistage reaction stepwise. In addition, it is preferable that products of the same quality can be stably produced, that is, it is not necessary to consider the difference between lots, and the measurement accuracy required by clinical examination can be satisfied.

Furthermore, the dry multilayer film is preferably formed by adhering a porous film. Examples of the porous membrane include cellulose-based porous membranes such as nitrocellulose porous membrane, cellulose acetate porous membrane, cellulose propionate porous membrane, and regenerated cellulose porous membrane, polysulfone porous membrane, and polyethersulfone porous membrane. Examples thereof include a membrane, a polypropylene porous membrane, a polyethylene porous membrane, and a polyvinylidene chloride porous membrane. More preferred are a polysulfone porous membrane and a polyethersulfone porous membrane.
The method for adhering the porous film to the dry multilayer film is not particularly limited. For example, the porous film is moistened with 15 to 30 g of water per 1 m ^{2 of the} dry multilayer film, and the porous film is 3 to 5 kg / cm ² at room temperature. The porous film can be bonded to the dry multilayer film by applying pressure to the film.

Moreover, it is also preferable that fine particles of 100 μm or less are adhered to the dry multilayer film and used for the reagent layer. Examples of the fine particles include inorganic fine particles typified by metal oxides such as silica, alumina, zirconia, and titania, and organic polymer fine particles typified by polystyrene (PS) and polymethyl methacrylate (PMMA). More preferred are silica and polystyrene.
The method for adhering the fine particles to the dry multilayer film is not particularly limited. For example, an aqueous solution in which 1 to 10% of polyvinylpyrrolidone (PVP), polyisopropylacrylamide, or a mixture thereof is added to the mass of the fine particles is added. The method of apply | coating and drying can be mentioned.

Depending on the type of specimen to be described later, it is preferable to filter the specimen before supplying the specimen to the portion carrying the developer reagent. Any conventionally known filtration can be applied. That is, it is preferable that the multi-item measurement dry analysis element has a filtration part. As the filter medium contained in the filtration part, either of the following two is preferable.
(I) A water-insoluble substance having an equivalent circle diameter of 5 μm or less and a length equal to or longer than the equivalent circle radius.
(II) A fiber having an equivalent circle diameter of 5 μm or less.
The above filtration unit is preferable because red blood cells can be quickly and efficiently removed from whole blood, particularly when whole blood is used as a specimen. In addition, it is preferable that plasma can be supplied to the reagent after removing red blood cells from whole blood without operating a special device, and as a result, the time until detection can be shortened.
More preferably, the combination of the fiber of (II) whose equivalent circle diameter is 5 μm or less and the porous membrane does not leak red blood cells even when the amount of whole blood is large, and a sufficient amount of plasma is obtained as a reagent. It is preferable that the fiber having an equivalent circle diameter of 5 μm or less is a glass fiber.
Hereinafter, the filter medium will be described in more detail.

The equivalent circle diameter in the present invention is a so-called equivalent diameter and is a term generally used in the field of mechanical engineering. When an equivalent circular pipe is assumed for a pipe having an arbitrary cross-sectional shape (in the present invention, it corresponds to a water-insoluble substance, fiber, or glass fiber), the diameter of the equivalent circular pipe is referred to as an equivalent diameter. , A: cross-sectional area of the pipe, p: wetting length (peripheral length) of the pipe, it is defined as deg = 4A / p. When applied to a circular tube, this equivalent diameter corresponds to the circular tube diameter. The equivalent diameter is used to estimate the flow or heat transfer characteristics of the pipe based on the data of the equivalent circular pipe, and represents the spatial scale (typical length) of the phenomenon. The equivalent diameter is deq = 4a ² / 4a = a for a regular square tube with one side a, and deq = 2h for a flow between parallel flat plates with a path height h. These details are described in “Mechanical Engineering Encyclopedia” (edited by the Japan Society of Mechanical Engineers, 1997, Maruzen Co., Ltd.).
The equivalent circle radius is calculated in the same manner as the equivalent circle diameter.

  Examples of water-insoluble substances include silicon, glass, polystyrene (PS), polyethylene terephthalate (PET), polycarbonate (PC), polyimide known by trademarks such as Kevlar, glass fiber, glass fiber filter paper, polyethylene terephthalate ( PET) fiber and polyimide fiber.

  Examples of the fiber include glass fiber, glass fiber filter paper, polyethylene terephthalate (PET) fiber, and polyimide fiber.

The porous membrane preferably has a pore size of 0.2 to 30 μm, more preferably 0.3 to 8 μm, more preferably about 0.5 to 4.5 μm, and particularly preferably 0.5 to 3 μm. .
The porosity is preferably high, specifically, the porosity is preferably from about 40% to about 95%, more preferably from about 50% to about 95%, still more preferably from about 70% to about 95%. It is a range.
Examples of porous membranes include conventionally known polysulfone membranes, polyethersulfone membranes, fluorine-containing polymer membranes, cellulose acetate membranes, nitrocellulose membranes and the like. A polysulfone membrane and a polyethersulfone membrane are preferable.
Further, the surface of which is hydrolyzed with a hydrophilic, hydrophilic polymer or activator can be used. As the hydrolysis method, hydrophilic polymer, activator, and the like applied to the hydrophilization treatment, methods and compounds that are usually used in the hydrophilization treatment can be used.

A polymer porous body can also be used as the filter medium. That is, a porous polymer body is installed in the flow path before supplying the specimen to the portion carrying the developer reagent. This is preferable because a solid component unnecessary for detection can be removed from the specimen and the specimen can be supplied to the reagent.
Examples of the polymer porous material include a polysulfone porous membrane, a polyethersulfone porous membrane, a fluorine-containing polymer porous membrane, a cellulose acetate porous membrane, a nitrocellulose porous membrane, a polystyrene porous fine particle, and a polyvinyl alcohol porous material. Fine particles. Preferred are a polysulfone porous membrane and a polyethersulfone porous membrane.

Further, instead of the above-mentioned filter medium, it is possible to remove a solid component unnecessary for detection by imprinting a space in the flow path itself, and supply the specimen to the reagent.
As an engraving method, there is a method in which a pattern formed using a photoresist on a silicon wafer is used as a mold, and a resin is poured into the pattern to mold (molding method). By marking a space in the flow path itself, a shape for removing a solid component unnecessary for detection can be formed in the flow path space, and a solid component unnecessary for detection can be removed. The shape formed by engraving is not limited to a cylindrical shape, and may be a prismatic shape or a hemispherical shape. Further, the shape formed by engraving preferably has an equivalent circle diameter of 5 μm or less, and the aforementioned (I) equivalent circle diameter is 5 μm or less and the length is equal to or longer than the equivalent circle radius. A water-insoluble substance can also be formed in the flow path by the method.
As a method for marking a space in the flow path itself, the above-described technique can be used as a fine processing technique in the flow path.

In addition, for example, a micro-pillar or nano-pillar shaped into a columnar shape using a processing technique such as microfabrication technology or μTAS is loaded with the color developing reagent. You may arrange | position and use for the flow path before supplying the sample to a part. There are various methods for making micro-pillars and nano-pillars. Silicon wafers can be left in the form of pillars by exposure and etching, or can be removed by pressing them onto the resin using a concave mold. An imprinting method in which columnar protrusions are formed on the surface may be used.
Furthermore, it is not necessary to limit the shape to a pillar shape. For example, a structure having an equivalent circle diameter of 5 μm or less may be created by using an optical modeling technique using a photocurable resin or the like.
Any of the materials used for the water-insoluble substance can be used as the shape to be arranged in the flow path before supplying the specimen to the portion carrying the developer reagent.
At this time, if a plurality of structures having an equivalent circle diameter of 5 μm or less are created and a structure that cross-links between the structures is created, more mechanical strength is imparted and both filtration performance and mechanical strength are satisfied. Can be made. Examples of the shape of the structure include a structure in which the pillars are cross-linked, a structure in which the fibers are cross-linked, a mesh structure having a grid pattern, checkered pattern, and honeycomb, and a cross-linked body thereof.

  Centrifugation may also be used to remove red blood cells from whole blood. When centrifugal separation is used, the multi-item dry analytical element itself or a part thereof has a form in which a centrifugal separator can be used, and the plasma is separated. Any form may be used as long as it can be guided to the carrying part.

The sample is injected from the inlet of the multi-item measurement dry analysis element. As long as it is possible to inject the specimen, any form may be used. For example, the flow path may be directly connected to the outside of the multi-item measurement dry analysis element.
Hereinafter, although an example of the preferable aspect of a multi-item measurement dry analysis element is demonstrated using FIG. 1, FIG. 2, this invention is not limited to this.
The specimen is injected from the inlet A3 of the multi-item measurement dry analysis element A100. The injected specimen passes through the channel A1 and is guided to the portion A2 carrying the color reaction reagent. As described above, the filter medium A6 can be arranged in the channel A1 depending on the type of the specimen, or a polymer porous body can be arranged, or a space is imprinted in the channel A1 itself. Can do. The developing reaction reagent A7 is arranged in the portion A2 carrying the developing reaction reagent. In FIG. 2, A1, A2 and A3 are produced using the microfabrication technology for the lower material A5, but as described above, the configurations of A1, A2 and A3 are produced, and the lower lid is attached instead of the lower material A5. It may be provided and assembled.

Examples of the material for the multi-item measurement dry analysis element include the same materials as those in the flow path. The preferred range is also the same.
The shape and size of the multi-item measurement dry analytical element may be any shape and size as long as they are easily held by hand. Specifically, for example, a shape having a rectangular shape with one side of the bottom surface of about 10 to 50 mm and a thickness of about 2 to 10 mm is preferable.
When assembling the multi-item measurement dry analysis element, the same technique as the joining technique used when connecting the portion carrying the color developing reagent and the flow path can be used.

  The specimen moves within the multi-measurement dry analytical element, that is, the movement from the flow path to the portion carrying the color reaction reagent includes methods using pressure, capillary action, etc. It is preferable to use pressure, particularly negative pressure.

The multi-item measurement dry analysis element can be loaded into a blood collection device to form a blood collection unit. Hereinafter, the blood collection unit will be described.
[Blood collection unit]
The blood collection unit contains a blood collection device having two or more parts slidable with respect to each other and the multi-item measurement dry analysis element. The element is loaded into the device and slides while maintaining a substantially airtight state. By this, a sealed space is defined so that pressure can be reduced. The blood collection unit has two or more parts that are slidable with respect to each other, the multi-item measurement dry analysis element can be loaded into a blood collection instrument, and can be decompressed internally by sliding while maintaining a substantially airtight state. Any shape and size may be used as long as it is possible to define a sealed space. It is preferable that the range is easy to hold by hand and easy to operate.
The blood collection unit defines a sealed space in the interior so that the pressure can be reduced, so that the collected whole blood quickly enters the flow path of the multi-measurement dry analytical element and reaches the portion carrying the color reaction reagent. Can lead to.
As the material of the blood collection unit, the same material as that in the flow path can be exemplified. The preferred range is also the same.
When assembling the blood collection unit, the same technique as the joining technique used when connecting the portion carrying the color developing reagent and the flow path can be used.

The blood collection device of the blood collection unit preferably has a puncture needle having a diameter of 100 μm or less and a tip angle of 20 degrees or less. By being in the above range, puncture can be performed smoothly, and pain during blood collection can be reduced, which is preferable.
As a method for joining the blood collection unit and the puncture needle, the same technique as the joining technique used when the portion carrying the color reaction reagent and the flow path are connected can be used.
The puncture needle is a hollow needle that collects blood from a blood vessel and introduces whole blood into the flow path of the multi-item measurement dry analysis element by decompression by sliding the blood collection unit. The puncture needle may be, for example, a normal injection needle as long as the above range is satisfied, but may be small in terms of micro blood sampling. It is also preferable to reduce pain during blood collection by narrowing the tip of the needle. Moreover, it can also be created using the above-described microfabrication technology.
The material constituting the puncture needle is usually a metal, and examples thereof include materials used as so-called injection needles such as stainless steels, nickel-titanium alloys, and tungsten. Moreover, it is also possible to use resins such as the above-mentioned plastics as a material constituting the multi-item measurement dry analysis element. Specific examples include PCO, PS, PC, PMMA, PE, PET, PP, and PDMS.

Although an example of the preferable aspect of a blood collection unit is demonstrated using FIG. 3, FIG. 4, this invention is not limited to this.
The multi-item measurement dry analysis element A100 is loaded into the blood collection device B1 from the direction C1 to form a blood collection unit B100. After loading, the puncture needle B2 is inserted into a human or an animal to collect whole blood D. As described above, a part of the blood collection device is slid in the direction C2, the pressure inside is thereby reduced, and the collected whole blood D enters the flow path A1 of the multi-item measurement dry analysis element A100, and further the color development. The reaction is carried to the part A2 carrying the reaction reagent. After the reaction, the multi-item measurement dry analysis element A100 can be removed from the blood collection device B1 and used for detection. The multi-item measurement dry analysis element A100 is removed from the blood collection device B1 in the direction C1, that is, in the same direction as when loading, or removed from the other side of the blood collection device B1, or the direction opposite to the direction C1, ie, the same as when loading. Any of the modes removed from the side may be used.
Further, when peripheral blood is taken out by puncturing a fingertip, elbow, heel, etc. with a lancet or the like and used for examination, a puncture needle is not necessary for a blood collection device of the blood collection unit. Any structure having a hollow structure and having a function of guiding blood to the analysis element may be used.

[Sample]
Samples to be used for the above multi-item dry analysis elements are used in human and other animal blood, body fluids and urine, environment-related substance testing fluids, agriculture, fisheries and food testing fluids, and natural science research. Liquid. Examples of the environment-related substance inspection liquid include fresh water, seawater, and a soil extract. Examples of liquids for agriculture, fisheries and food inspection include agricultural products and agricultural products extracts, marine products and marine products extracts, processed foods of agricultural products and / or marine products, and processed food extracts obtained by processing agricultural products and / or marine products. it can. Liquids used in natural science research include liquids used in chemical, biology, geology, physics research, and the like.

Hereinafter, the schematic configuration of the measurement apparatus when an area sensor is used will be described with reference to FIG.
The measurement apparatus 100 includes a multi-item measurement dry analytical element installation unit 1 for installing a sample to be measured, a light source 2 using a light emitting element such as a halogen lamp that irradiates light on the sample, and light emitted from the light source 2. Variable light section 3 that changes the intensity of light, wavelength variable section 4 that changes the wavelength of light emitted from the light source 2, lenses 5a and 5b that collimate and collect the light emitted from the light source 2, and the specimen The lens 5c that collects the reflected light from the lens, the area sensor 6 as a light receiving element that receives the reflected light collected by the lens 5c, and controls each part and receives the state of the light variable unit 3 and the area sensor 6. And a computer 7 that obtains a measurement result corresponding to the amount of light and outputs the result to a display or the like. Here, the computer 7 is configured to control each unit, but a computer that performs overall control of each unit may be prepared separately.

  The multi-item measurement dry analysis element installation unit 1 is provided with a multi-item measurement dry analysis element. What is actually used for the measurement is a portion (hereinafter referred to as “reagent carrying portion”) of the multi-item measurement dry analytical element that carries the color reaction reagent that has reacted with the specimen.

  The light variable section 3 is irradiated from the light source 2 to the specimen by mechanically inserting and removing a metal mesh plate material such as stainless steel with a hole and a neutral density filter such as an ND filter between the light source 2 and the specimen. It changes the intensity of light. In the initial setting, the neutral density filter is inserted between the light source 2 and the specimen. In the following description, the metal mesh is a stainless steel mesh. Moreover, a stainless steel mesh plate with a hole and a neutral density filter such as an ND filter may be manually inserted and removed.

  The wavelength variable unit 4 changes the wavelength of light emitted from the light source 2 to the specimen by mechanically inserting or removing one of a plurality of types of interference filters between the light source 2 and the specimen. In this embodiment, the wavelength variable unit 4 is installed between the light variable unit 3 and the multi-item measurement dry analytical element installation unit 1. However, it is installed between the light source 2 and the light variable unit 3. Also good. Further, a plurality of types of interference filters may be manually inserted and removed.

  The area sensor 6 is a solid-state imaging device such as a CCD, and when the reagent of the reagent carrier in the multi-item measurement dry analysis element installed in the multi-item measurement dry analysis element installation unit 1 reacts with a sample such as blood. The light reflected by the light emitted from the light source 2 is received, and the received light is converted into an electrical signal and output to the computer 7. The area sensor 6 can receive the light reflected from the reagent carrying part on a surface basis. Therefore, it is possible to measure each reagent area simultaneously, that is, for a plurality of items.

  The computer 7 converts an electrical signal corresponding to the amount of received light output from the area sensor 6 into an optical density value based on calibration curve data stored in a built-in memory or the like, and uses the optical density value as a sample. The content of various components contained in the product is obtained and output to a display or the like. When measuring a plurality of items, the computer 7 extracts an electrical signal corresponding to the amount of received light output from the area sensor 6 for each of the plurality of areas of the reagent carrying unit, and calculates the content of the component contained in the sample for each of the plurality of areas. Ask for. Further, the computer 7 controls the light variable unit 3 and the wavelength variable unit 4 according to the amount of reflected light from the sample received by the area sensor 6 and the type of reagent to be reacted with the sample, and the amount of light from the light source 2 is controlled. Change the wavelength or change its wavelength.

In the measuring apparatus 100 configured as described above, when the amount of light reflected from the specimen is so small that it does not fall within the dynamic range of the area sensor 6, the light variable section 3 is made of stainless steel mesh between the light source 2 and the specimen. The plate material or the ND filter is removed, and the intensity of light emitted from the light source 2 is increased. As a result, the amount of reflected light from the specimen increases, and the amount of reflected light falls within the dynamic range of the area sensor 6. For this reason, even if the dynamic range of the area sensor 6 is narrow, the reflected light can be received with high accuracy, and the measurement accuracy of the content of the component contained in the specimen is improved.

  For example, in the case of using a reagent carrying unit including four types of reagents A, B, C, and D, the measuring apparatus 100 obtains the reflected light amount from each area including the reagents A to D, and the reflected light amount. If any of the above does not fall within the dynamic range of the area sensor 6, the light variable section 3 inserts and removes a stainless mesh plate or ND filter at regular intervals. In addition, since the wavelengths of light reflected from each area are different, the wavelength variable unit 4 switches a plurality of interference filters according to the wavelength.

  For example, the amount of reflected light from the area including A and B is so small that it does not fall within the dynamic range of the area sensor 6, and the amount of reflected light from the area including C and D falls within the dynamic range of the area sensor 6. A case will be described in which the wavelengths of light emitted when the reagents of -D react with blood differ.

  In this case, in the measuring apparatus 100, the light source 2 irradiates the reagent carrying portion with light, the reflected light from each area of the slide is received by the area sensor 6, and the reflected light amount from each area falls within the dynamic range of the area sensor 6. It is determined by the computer 7 whether or not Here, since the amount of reflected light from the area including A and B is so small that it does not fall within the dynamic range of the area sensor 6, the computer 7 controls the light variable unit 3 after light is emitted from the light source 2 for a certain period of time. Then, the ND filter is removed from between the light source 2 and the specimen. After irradiating light for a certain time in this state, the computer 7 controls the light variable section 3 to insert an ND filter between the light source 2 and the specimen. By repeating this operation, a plurality of types of measurement components can be accurately measured with one multi-item measurement dry analysis element.

  The computer 7 controls the optical variable unit 3, while controlling the wavelength variable unit 4 according to the types of reagents A to D, and sequentially switches the four types of interference filters. While the optical variable unit 3 removes the ND filter, the wavelength variable unit 4 alternately switches between the interference filter corresponding to the reagent A and the interference filter corresponding to the reagent B, and the optical variable unit 3 inserts the ND filter. During the operation, the interference filter corresponding to the reagent C and the interference filter corresponding to the reagent D are switched alternately. Thereby, even when the wavelengths of light emitted from a plurality of types of components contained in the specimen are different from each other, the content of the plurality of types of measurement components contained in the specimen can be measured with one multi-item measurement dry analytical element. it can.

  The measuring apparatus 100 can measure with high accuracy even with a CCD having a narrow dynamic range by changing the intensity of light from the light source 2, but the control of the computer 7 without changing the intensity of light. By changing the exposure time (received time of reflected light) in the CCD, it is possible to perform highly accurate measurement as described above.

  In this embodiment, the sample is irradiated with light from the light source 2 and the content of the component contained in the sample is obtained from the reflected light. However, the content of the component contained in the sample from the transmitted light that has passed through the sample. You may ask for.

  In this embodiment, reflected light from the specimen is received using an area sensor such as a CCD. However, the present invention is not limited to the area sensor, and a line sensor may be used.

In the CCD used in this embodiment, light receiving portions such as photodiodes are arranged vertically and horizontally on a semiconductor substrate at predetermined intervals, and the light receiving portions included in adjacent light receiving portion rows are mutually within the light receiving portion row. It is desirable to use a so-called honeycomb type CCD that is shifted in the direction of about ½ row of the pitch between the light receiving portions in FIG.

  In the above description, the measurement apparatus 100 changes the light intensity in real time according to the amount of reflected light from the specimen. However, in the sequence set in advance according to the measurement component included in the specimen, the measurement component 100 The content may be measured. The operation in this case will be described below.

  When the reagent carrying unit is installed in the multi-item measurement dry analysis element installation unit 1 and the measurement item is set, the measurement apparatus 100 starts measurement with a pattern corresponding to the measurement item. First, the computer 7 selects light intensity used for measurement from a plurality of types of intensity, and irradiates the specimen with light of the selected intensity. When the area sensor 6 receives the reflected light reflected from the specimen, the computer 7 outputs a measurement result corresponding to the amount of the reflected light received by the area sensor 6 and the intensity of the selected light. By this series of operations, it is possible to accurately measure the measurement component contained in the specimen.

  When changing the exposure time of the CCD without changing the light intensity, the measuring apparatus 100 has a reagent carrying unit installed in the multi-item measurement dry analytical element installation unit 1 and sets the measurement item. Start measurement with the corresponding pattern. First, the computer 7 irradiates the specimen with light. Then, the area sensor 6 receives the reflected light reflected from the specimen for an exposure time selected by the computer 7 from a plurality of types of exposure times. Finally, the computer 7 outputs a measurement result corresponding to the amount of reflected light received by the area sensor 6 and the selected exposure time. By this series of operations, it is possible to accurately measure the measurement component contained in the specimen.

  The measuring apparatus 100 is not limited to obtaining light from the light source 2 described above to irradiate the reagent-carrying portion and obtaining the content of the component contained in the specimen from the reflected light or transmitted light. The content of the component contained in the specimen may be obtained by detecting light such as fluorescence emitted from the reagent carrying part when the reagent carrying part is irradiated with light from the light source. The content of the component contained in the specimen is detected by detecting light emitted by chemiluminescence or the like emitted from the reagent-carrying part in a state where no light is applied to the reagent-carrying part without using the light source 2 You may ask for it.

  The present invention will be described below with reference to examples, but the present invention is not limited thereto.

[Example of apparatus] Configuration of measuring apparatus A photometric system having an optical arrangement shown in FIG. 5 was prepared. Specifically, each member was prepared as follows.
Optical system: inverted stereo microscope.
The following two magnifications are available at the CCD light receiver.
0.33 times; 33μm / pixel at CCD
1 ×; 10 μm / pixel at CCD part Light source 2; Luminer Ace LA-150UX manufactured by Hayashi Clock Industry
Wavelength variable section 4 (interference filter); monochromatic at 625 nm, 540 nm, and 505 nm, respectively, light variable section 3 (dark filter); glass filter manufactured by HOYA ND-25
And homemade filter area sensor 6 (CCD) with holes in stainless steel plate; 8-bit monochrome camera module XC-7500 manufactured by Sony Corporation
Computer 7 (data processing (image processing)); Image processing device LUZEX-SE manufactured by Nireco Corporation.
Means for calibrating the reflection optical density: The following six types of standard density plates (ceramic specifications) made by Fuji Equipment Co., Ltd. are prepared.
Standard density plate; A00 (reflection optical density to 0.05),
A05 (0.5)
A10 (1.0),
A15 (1.5),
A20 (2.0),
A30 (3.0).

[Example 1]
The shape of the rubber portion into which the puncture needle of Terumo 10 mL vacuum blood collection tube (inner diameter 13.5 mm) was inserted was left as it was, and the portion of the resin tube was cut with a cutter. With the puncture needle inserted into the rubber part of the cut blood collection tube to allow air to enter and exit, the piston part of a Terumo syringe was inserted, and the puncture needle was pulled out to a distance of about 10 mm from the rubber part. In that state, the piston was pulled an arbitrary distance to reduce the pressure, and the piston was fixed so as not to move (1). Whole blood previously collected using heparin lithium as an anticoagulant is injected into another Terumo syringe, a puncture needle is attached to the tip, and the puncture needle is attached to the rubber part of (1) to which the piston is fixed. The whole blood volume that was inserted and collected in (1) by decompression was determined by the gravimetric method. As shown in Table 1 and FIG. 6, it was found that whole blood could be collected in an amount corresponding to the reduced volume when the piston was pulled to reduce the pressure. Therefore, the blood collection unit can be loaded with a multi-item measurement dry analytical element in a blood collection device, and can be sealed to define a sealed space in which pressure can be reduced by sliding while maintaining a substantially airtight state. It was found that whole blood could be introduced into the item measurement dry analytical element.

[Example 2]
A multi-item measurement dry analysis element 20 of about 24 mm × 28 mm made of polystyrene resin (PS) shown in FIG. 7 is prepared, and the base material 22 of the multi-item measurement dry analysis element 20 has a width of 2 mm, a length of 10 mm, and a depth of 2 mm. The glass fiber filter paper (manufactured by Whatman; GF / D) 27 and the polysulfone porous membrane (PSF made by Fuji Photo Film) 28 for red blood cell capture / plasma extraction are placed in the flow path 23 of FIG. The color developing reagent 24 has a width of 5 mm, a length of 5 mm, and a depth of 2 mm, and Fuji Dry Chem mount slides GLU-P and TBIL-P as the color developing reagent 24. (Fuji Photo Film Co., Ltd.) is cut into a width of 2 mm and a length of 4 mm, and loaded so that GLU-P is on top and TBIL-P is on bottom. And to keep the airtightness-watertight bonded with tape.
Next, 100 μL of plain blood collected in the tube 25 on the upper glass fiber filter paper 27 side is inserted and allowed to stand for 10 to 20 seconds to spread the whole blood on the glass fiber filter paper. A Terumo syringe was attached to the tube 26 on the side opposite to the side, and suction was performed slightly. Plasma extracted by filtration leaks from the polysulfone porous membrane 28 and is dropped onto the dry chem mount slide, and the dry chem mount slides GLU-P and TBIL-P (hereinafter also referred to as GLU-P and TBIL-P slides) gradually. Color development was started (FIGS. 8 to 10). It took 30 seconds to extract plasma and drop it onto the mount slide after the plain whole blood was injected.

The color development of the GLU-P and TBIL-P slides was simultaneously imaged with a CCD camera using the optical system of [Example of apparatus], and the images obtained using LUZEX-SE were processed, and GLU-P and TBIL- The average amount of light received at the center of the image on the P slide was calculated and converted to an optical density, and the glucose and total bilirubin concentrations in the specimen were determined. When processing the image captured by the CCD camera with LUZEX-SE, about the central part of the GLU-P and TBIL-P images,
The amount of received light in a range of 1 mm in length × 2 mm in width was calculated by image processing. At this time, since the magnification of 0.33 times of the optical system was used, the pixels were measured with 30 pixels in length × 60 pixels in width, that is, with 1800 pixels. In order to compare whether the results measured using a CCD camera are correct, the glucose and total bilirubin concentrations in the specimen were determined using an automatic clinical test apparatus 7170 manufactured by Hitachi, Ltd. The results are shown in Table 2. At this time, since the measurement wavelength differs between the GLU-P and TBIL-P slides, photometry was performed by sequentially changing the wavelength of the interference filter every 5 seconds as shown in Table 3.
Thus, it was found that the multi-item dry analytical element of the present invention is easy to operate and can be quickly measured. Here, the dry chemistry reagent for two items was used as the developing reaction reagent, but the number of items can be increased.

[Example 3]
[Measurement with standard density plate]
The relationship between the optical density and the amount of reflected light received was obtained using a standard density plate using light monochromatic to 625 nm in the optical arrangement of the apparatus example. The region of received light amount “range of received light amount of 200 to 50” that can be measured accurately with an 8-bit monochrome CCD was defined as the range of the calibration curve, and the optical density was determined as follows.
(1) Using a standard density plate having an optical density of approximately 0, a light-reducing filter is inserted so that the amount of received light is around 200, and the amount of light emitted from the light source is adjusted. Using six types of standard density plates, A calibration curve was created by determining the relationship between the optical density and the amount of reflected light. When a perforated stainless steel plate was used as the neutral density filter, the amount of light emitted at the sample site was 96 μW / cm ² .
(2) Only the neutral density filter was removed in the state of the optical system in (1), and the relationship between the optical density and the amount of reflected light was obtained using six types of standard density plates, and a calibration curve was created. When the perforated stainless steel plate used as the neutral density filter was removed, the amount of light emitted from the sample portion was 492 μW / cm ² .
(3) The region until the amount of reflected light received when measured under the condition (1) is less than 50 (the reflection optical density 0 to 0.9 in FIG. 11) is the region X, and the condition (2) above. A region until the amount of reflected light received when measured is less than 50 and does not overlap with region X (reflected optical density of 0.9 to 1.8 in FIG. 11) is defined as region Y.
(4) Using the calibration curve a measured under the condition (1) in the range of the region X and the calibration curve b measured under the condition (2) in the range of the region Y, the reflection optics of the sample described later is used. Concentration was measured.

  Subsequently, it was measured by reflection photometry at N = 10 using a standard density plate. The standard deviation of the reflection optical density was determined. When the density plate A05 having a low optical density is used, the area X is measured using the neutral density filter according to the condition (1). When the density plates A10 and A15 having a high optical density are used, the area is measured. Since Y, the neutral density filter was removed in accordance with the condition (2). As a result, the standard deviation of reflection optical density (SD of OD) of 10 / 10,000 or less was achieved in any of A05, A10, and A15, and measurement was possible with high accuracy. The magnification of the optical system at the time of measurement was 0.33, and the amount of received light was calculated by performing image processing on the diameter of 5 mm at the center of the image of the standard density plate imaged by the CCD camera. This is a circle with a radius of 75 pixels and was therefore measured with a pixel count of 17662. Note that the time required for the measurement was 1 second in total for photometry and image processing.

Using the optical system shown in FIG. 5, an experiment was performed to increase the accuracy when quantifying a plurality of items simultaneously using a plurality of interference filters. In this experiment, dry clinical examination used for Fuji Photo Film Co., Ltd. Fuji Dry Chem Mount Slide GLU-P (measurement wavelength: 505 nm, measurement component: glucose) and TBIL-P (measurement wavelength: 540 nm, measurement component: total bilirubin). Each test piece of reagent is cut to about 2 mm × 4 mm, and loaded into a cell made of 5 mm × 5 mm transparent resin. From the upper part of the test piece, control serum (in this case, L 4 μL of 2 types of H) were added dropwise, and at room temperature, the measurement component in the serum was reacted with the reagent to develop a color.

  At this time, in order to calibrate the reflection optical density from the measurement component, the calibration object developed by stepwise solid exposure of the black and white photographic paper was cut into about 1.5 mm × 2 mm and four sheets (level 1 to level) 4) These were placed side by side and placed in the same field of view as the above two test pieces (CCD imageable range), and imaged with a CCD using light monochromated by an interference filter. In this case, the computer 7 receives the reflected light from the calibration object together with the reflected light from the other specimen, and performs an operation of correcting the optical density of the component contained in the other specimen based on the light quantity. In this experiment, the light quantity and wavelength of light irradiated on the slide were sequentially changed in the order shown in Table 4 below, and the reflection optical density of the calibration object was shown in Table 5.

  For the measurement component having a reflected light quantity of 50 to 200 received by the CCD when using light with wavelengths of 505 nm and 540 nm with the neutral density filter inserted, use the calibration curve a shown in FIG. The reflected optical density is obtained from the reflected light amount, and for the measurement component having a reflected light amount of less than 50, the reflected light is received using the calibration curve b shown in FIG. 11 from the reflected light amount received by the CCD with the neutral density filter removed. The concentration was determined. From the reflection optical density when the glucose and total bilirubin thus obtained are colored, and the calibration curve data stored in advance in the computer 7 in which the reflection optical density is associated with the content of the measurement component, Serum concentrations of glucose and total bilirubin were calculated. The calculation results are shown in Table 6 below.

  As shown in Table 6, since the actual measurement value and the control serum standard value almost coincide, even with a CCD having a narrow dynamic range, the content of the measurement component in the serum can be accurately measured. Prove that you can. In addition, since two measurement components are measured at the same time, it was possible to measure efficiently compared to the case where the two slides of GLU-P and TBIL-P were separately divided as in the prior art. . In this embodiment, only two measurement components are measured, but it is possible to simultaneously measure the density of two or more measurement components as long as they are within the CCD imaging range.

[Study on the number of pixels]
In the optical system shown in FIG. 5, an optical image of the standard density plate A05 was picked up with N = 10 using light monochromatic to 625 nm, and the standard deviation of the reflected optical density of the density plate was obtained. When obtaining the reflection optical density, the reflection optical density was calculated by changing the region near the center of the imaged density plate, and the dependence of the standard deviation of the reflection optical density on the photometric area was obtained. The results are shown in Table 7, Table 8 and FIG.
The actual photometric area and the CCD pixel area differ depending on the magnification of the lens, but when the number of pixels in the photometric area exceeds 1000, the standard deviation of the optical density becomes 10 / 10,000 or less, and the measurement is performed accurately. I was able to. In addition, a pixel here is a pixel, and similarly, the number of pixels is the number of pixels.

[Example 4]
When a dry multilayer film was used as a color developing reagent for a multi-analytical dry analytical element, it was considered that the unevenness of the multilayer film photometric surface affects the amount of reflected light and the reproducibility of the reflected optical density. Using a multilayer film with different surface irregularities, the simultaneous reproducibility of the reflection optical density was measured while changing the photometric size. For comparison, the same measurement was performed on a standard density plate made of ceramic with a smooth surface. Fuji Dry Chem Mount Slide CRP-S manufactured by Fuji Photo Film Co., Ltd. was used as a multilayer film with large surface irregularities, and Fuji Dry Chem Mount Slide BUN-P manufactured by Fuji Photo Film Co., Ltd. was used as a multilayer film with small surface irregularities. . CRP-S has a large unevenness on the reflection surface due to the texture of the cloth affixed to the side opposite to the photometry surface, and BUN-P reflects a porous film on the intermediate layer. The unevenness of the reflection surface when measuring light is small. In addition, the standard density plate A05 (reflective optical density 0.5) manufactured by Fuji Kikai Kogyo Co., Ltd. was used as the standard density plate made of ceramic.
The same optical system as that shown in FIG. 5 was used, and the magnification at the CCD light receiving portion was 1 × (10 μm / pixel at the CCD portion).

  Table 9 and FIG. 13 show the standard deviation of the reflected optical density when the reflected optical density is measured 10 times while changing the photometric diameter from 0.2 mm to 3 mm. When the photometric diameter was 3 mm, it was found that the standard deviation could be measured with a precision of 10 / 10,000 or less for any multilayer film. When the diameter of photometry is reduced, the standard deviation increases, and in CRP-S, the photometric diameter exceeds 10 / 10,000 when the photometric diameter is 1 mm, whereas with BUN-P using a porous film, the photometric diameter is 1 mm. It was 10 / 10,000 or less. It was found that the use of a porous film can reduce the surface unevenness of the reflection surface when performing reflection photometry, and the measurement can be performed with higher accuracy. In the measurement using the standard density plate A05 having extremely small surface irregularities, the number of pixels in the photometric area is less than 1000 and the standard deviation exceeds 10 / 10,000 at the photometric diameter of 0.2 mm. When 1 mm, the standard deviation was 2.4 / 10,000. It is a point to improve the measurement accuracy that the unevenness of the reflective surface at the time of reflection photometry, such as a porous film or fine particles, is smaller than a cloth that is practically used for Fuji Dry Chem Mount Slide CRP-S. I understood.

[Example 5]
We observed how red blood cells in whole blood were captured by glass fiber, which is one of the fibers used as a filter medium for multi-analytical dry analytical elements. Whole blood was collected from healthy men with a vacuum blood collection tube using lithium heparin as an anticoagulant. The Hct value at this time was 45%. 10 μL of this whole blood was dripped onto a glass fiber filter paper GF / D (glass fiber diameter of about 3 μm or less) manufactured by Whatman Co. at room temperature, and a 0.1 mol / L phosphate buffer (pH 7.5) containing 1% glutaraldehyde. In 4), a glass fiber filter paper in which whole blood was dripped was quickly put, and allowed to stand at room temperature for 2 hours to harden red blood cells. Then, the glass fiber filter paper is immersed in a water / t-butanol mixture, and the water / t-butanol mixture ratio is gradually changed to finally replace with t-butanol, and then left in a refrigerator for about 1 hour to freeze. It was. The frozen t-butanol solution containing glass fiber filter paper was placed in a lyophilizer to remove the solvent. The dried glass fiber filter paper with whole blood dripped in this way was observed with a scanning electron microscope to obtain a photograph with a magnification of 1000 times. A photograph is shown in FIG. In the photograph of FIG. 14, the full-scale width of the photograph is 120 μm. It was found that erythrocytes were captured by glass fibers having a diameter of about 3 μm or less.
For comparison, a similar experiment was conducted using a glass fiber filter paper having a diameter of about 8 μm and a diameter of about 10 μm, and an acetyl cellulose fiber filter paper having a diameter of about 15 μm as filter media. It was found that erythrocytes could not be captured with a glass fiber having a diameter of about 8 μm. It was found that red blood cells could not be captured at all with glass fibers having a diameter of about 10 μm and acetylcellulose fibers having a diameter of about 15 μm.
Therefore, by using a fiber with a specific equivalent circle diameter, that is, a water-insoluble substance, as a filter medium for a multi-analytical dry analytical element, when using whole blood as a specimen, erythrocytes can be efficiently removed from whole blood quickly. It was found that it can be removed well. Furthermore, since it is not necessary to operate a special device to remove red blood cells from whole blood, it has been found that plasma can be rapidly supplied to the reagent and the time to measurement can be shortened.

It is a schematic diagram of one Embodiment of a multi-item measurement dry analysis element. It is a schematic diagram of one Embodiment of a multi-item measurement dry analysis element. It is a schematic diagram of one Embodiment in a blood collection unit. It is a schematic diagram of one Embodiment in a blood collection unit. It is a schematic diagram of one Embodiment in a measuring apparatus. It is a graph showing the relationship between a decompression volume and the amount of blood collection. It is the schematic which shows one Embodiment in the multi-item measurement dry analytical element of the aspect of Example 2. FIG. It is a photograph which shows one Embodiment in the multi-item measurement dry analytical element of the aspect of Example 2. FIG. In one Embodiment in the multi-item measurement dry analytical element of the aspect of Example 2, it is the photograph after inject | pouring whole blood. In one Embodiment in the multi-item measurement dry analytical element of the aspect of Example 2, after injecting whole blood, it is the photograph which shows that the color development reaction reagent started color development by attracting | sucking with a thermo syringe. It is a graph which shows the relationship between reflection optical density and reflected received light amount. It is a graph which shows the photometry area dependence of the standard deviation of reflected optical density. It is a graph which shows the photometry area dependence of the standard deviation of reflected optical density. It is a scanning electron micrograph after dropping whole blood on glass fiber and freeze-drying.

Explanation of symbols

A100 Multi-item dry analysis element A1 Flow path A2 Portion carrying A3 reaction reagent A3 Inlet A4 Upper cover A5 Lower material A6 Filter medium A7 Development reaction reagent E1 Joining direction E2 of upper cover Arrow E3 indicating the location of the filter medium Arrow B100 indicating the location of the color developing reagent B100 Blood collection unit B1 Blood collection device B2 Puncture needle C1 Loading direction C2 of the multi-item measurement dry analytical element D2 Sliding direction when depressurizing D Whole blood 100 Measuring device 1 Multi-item measurement dry analytical element Installation unit 2 Light source 3 Light variable unit 4 Wavelength variable unit 5a, 5b, 5c Lens 6 Area sensor 7 Computer 20 Multi-item measurement dry analysis element 21 Upper material 22 Lower material 23 Channel 24 Color reaction reagent 25 Tube (injection port)
26 Tube 27 Glass fiber filter paper 28 Polysulfone porous membrane

Claims (21)

  A multi-item measurement dry analysis element for use in a specimen testing method that uses an area sensor as a detector to obtain a measurement result from information of 1000 pixels or more per item and can simultaneously measure a plurality of items. The multi-item measurement dry analysis element has a flow path, a color reaction reagent, and a part carrying the color reaction reagent,
At least one of the width, depth, and length of the flow path is 1 mm or more,
And the width of the part carrying the color reaction reagent is at least twice the width of the flow path and / or the length of the part carrying the color reaction reagent is 0.4 times the length of the flow path The multi-item measurement dry analysis element according to claim 1, which is as described above.   The multi-item measurement dry analytical element according to claim 2, further comprising a filtration part containing a water-insoluble substance having an equivalent circle diameter of 5 µm or less and a length equal to or longer than the equivalent circle radius.   The multi-item measurement dry analytical element according to claim 2, comprising a filtration part containing fibers having an equivalent circle diameter of 5 µm or less.   The multi-item measurement dry analytical element according to claim 2, comprising a filtration part containing a fiber having a circle equivalent diameter of 5 µm or less and a porous membrane.   The multi-item measurement dry analytical element according to claim 2, comprising a filtration part containing glass fibers having an equivalent circle diameter of 5 µm or less and a porous membrane.   The multi-item measurement dry analytical element according to any one of claims 2 to 6, wherein a dry multilayer film is used as a reagent layer in a portion carrying the developer reagent.   The dry analysis element for multi-item measurement according to claim 2 or 3, wherein a dry multilayer film formed by adhering a porous film is used as a reagent layer in the portion carrying the developer reagent.   The dry analysis element for multi-item measurement according to claim 2 or 3, wherein a dry multilayer film to which fine particles having a diameter of 100 µm or less are adhered is used as a reagent layer in the portion carrying the color reaction reagent.   The multi-item measurement dry analytical element according to claim 2 or 3, wherein the portion carrying the developer reagent is a cell connected to the flow path.   The multi-item measurement dry analytical element according to claim 2 or 3, wherein the specimen is supplied to the reagent through the polymer porous body, and the dry multilayer film is used as a reagent layer in the portion carrying the color reaction reagent.   The multi-item measurement dry analysis according to claim 2 or 3, wherein the specimen is supplied to the reagent through the space marked in the flow path, and the dry multilayer film is used as a reagent layer in the portion carrying the color reaction reagent. element. In a multi-item measurement dry analytical element for use in a specimen testing method capable of measuring multiple items at the same time using a line sensor as a detector,
The multi-item measurement dry analysis element has a flow path, a color reaction reagent, and a part carrying the color reaction reagent,
At least one of the width, depth, and length of the flow path is 1 mm or more,
And the width of the part carrying the color reaction reagent is at least twice the width of the flow path and / or the length of the part carrying the color reaction reagent is at least 0.4 times the length of the flow path And
A multi-item measurement dry analytical element having a filtration part containing a water-insoluble substance having an equivalent circle diameter of 5 μm or less and a length equal to or longer than the equivalent circle radius. In a multi-item measurement dry analytical element for use in an analyte test method capable of measuring multiple items simultaneously using an electrochemical detector as a detector,
The multi-item measurement dry analytical element has a flow path, a reaction reagent, and a part carrying the reaction reagent,
At least one of the width, depth, and length of the flow path is 1 mm or more,
A multi-item measurement dry analytical element having a filtration part containing a water-insoluble substance having an equivalent circle diameter of 5 μm or less and a length equal to or longer than the equivalent circle radius.   A blood collection device having two or more parts slidable with respect to each other, and the multi-item measurement dry analysis element according to claim 2, wherein the element is loaded into the device, and the device is slid while maintaining a substantially airtight state. A blood collection unit characterized in that a sealed space is defined in the interior so that pressure can be reduced.   The blood collection unit according to claim 15, wherein the blood collection instrument has a puncture needle having a diameter of 100 µm or less and a tip angle of 20 degrees or less.   A blood collection device having two or more parts slidable with respect to each other and the multi-item measurement dry analysis element according to claim 13, wherein the element is loaded into the device, and sliding is performed while maintaining a substantially airtight state. A blood collection unit characterized in that a sealed space is defined in the interior so that pressure can be reduced.   The blood collection unit according to claim 17, wherein the blood collection instrument has a puncture needle having a diameter of 100 µm or less and a tip angle of 20 degrees or less.   The multi-item measurement dry analytical element according to claim 2, wherein a liquid for testing environment-related substances is used as a specimen.   The multi-item measurement dry analytical element according to claim 2, wherein a liquid for agriculture, fisheries or food inspection is used as a specimen.   The multi-item measurement dry analytical element according to claim 2, wherein a liquid used in natural science research is used as a specimen.