JP4657084B2 - Sample analysis disc - Google Patents

Description

  The present invention relates to a device used for detecting a chemical reaction between a sample and a reagent mainly in a clinical laboratory field.

In recent years, it has become possible to measure various substances due to advances in analysis, analysis, and inspection techniques. In particular, in the field of clinical testing, the development of measurement principles based on specific reactions such as biochemical reactions, enzyme reactions, or immune reactions has made it possible to measure substances in body fluids that reflect disease states.
Among them, a field of clinical examination called point-of-care testing (POCT) has attracted attention. In POCT, simple and quick measurement is the first, and efforts are being made to shorten the time from when a sample is collected until the test result is obtained. Therefore, the measurement principle and measurement device required for POCT are small, portable, and easy to operate based on a simple measurement principle.

Today, POCT-compliant measuring instruments are becoming practical due to the development of simple measurement principles and the accompanying advances in solid-phase technology, sensor device technology, sensor system technology, microfabrication technology, and microfluidic control technology. It's getting higher.
As such an analytical apparatus that can be used as a POCT-compatible measuring instrument, for example, Patent Document 1 proposes an apparatus that performs qualitative and quantitative analysis of a sample developed on a disk.

The measuring device using the technique described in Patent Document 1 can diagnose a disease by analyzing a sample such as blood. Here, FIG. 3 is a block diagram showing an analysis disk used in the conventional liquid sample analyzer. As shown in FIG. 3, a sample injection hole 104 and a channel 105 are provided in the disk 101, and a reagent 106 that changes optical characteristics (transmittance, color, etc.) reacts with the sample and is applied to the channel. Yes.
The analysis is performed by injecting a sample into the disk 101 from the sample injection hole 104 and mounting it on the analyzer.

FIG. 4 is a block diagram showing the analyzer in Patent Document 1. The configuration of the apparatus is similar to a so-called optical disk apparatus, and is a spindle motor 201 for rotating the disk 101, an optical pickup for irradiating the sample 106 developed in the disk 101 or the reagent 106 reacted with the sample 900 with a light beam. 212, a feed motor 213 for moving the optical pickup 212 in the radial direction of the disk 101, and the like.
The disk 101 mounted on the apparatus is rotated by the spindle motor 201, and the sample 900 is developed in the flow path 105 of the disk 101 by the centrifugal force, and simultaneously reacts with the reagent 106 applied in the flow path 105. After the reaction is completed, the reaction state of the reagent is detected by irradiating the sample 900 or the reagent 106 in the channel 105 with a light beam using the optical pickup 212 while rotating the disk and detecting the reflected light or transmitted light. And analyze.

In addition to the above-described disk configuration, for example, in order to react only the plasma component in blood with a reagent, for example, to remove blood cells by centrifugation or to dissolve and react a plurality of reagents sequentially. There has also been proposed a structure to which a function of freely moving and stopping the sample liquid is provided by providing a flow path connecting a plurality of chamber portions to which the reagent is applied and between the chamber portions (for example, Patent Document 2).
In the invention described in Patent Document 2, the movement and stop of the sample solution caused by the structure described in Patent Document 1 are based on the basic mechanism described below. FIG. 5 is a diagram for explaining the technique proposed in Patent Document 2. In FIG.

As shown in FIG. 5, the channel 302 exits from the lower side 301a when viewed from the direction of the centrifugal force of the upstream chamber 301 of the sample liquid flow, and viewed from the direction of the centrifugal force of the chamber. It is lifted to a position 302a above the level from the upper wall surface of the case, and then continues downward when viewed with reference to the direction of centrifugal force, and is connected to the downstream chamber 303 arranged at the end. The same flow path 304 is connected to the transmitted light measurement chamber 305.
It should be noted that the depth of the chamber is larger than the depth of the flow path. As a result, the sample solution that has moved in the flow path by capillary action is prevented from moving by the capillary action at the portion where the flow path is connected to the chamber, and as a result, the sample liquid can be stopped before the chamber. The sample liquid that has been stopped flows into the chamber (lower chamber 303) by applying centrifugal force by rotating the disk from this state.

As a structure that should be further noted here, as described above, when it is lifted up to a position 302a above the level from the upper wall surface when viewed with reference to the centrifugal force direction of the same chamber, then, when viewed with reference to the centrifugal force direction It is a feature in the arrangement of the flow path that it continues below.
Because of this feature, when a centrifugal force is applied, a siphon effect is exerted on the sample solution that has accumulated in the upstream chamber 301 and filled up to the front portion of the lower chamber of the flow path 302, and the upstream side passes through the flow path 302. Almost all of the sample liquid accumulated in the side chamber flows into the lower side chamber 303.

However, while the centrifugal force is acting, the sample liquid that has flowed into the lower chamber 303 also enters the flow path 304, but the same as when the liquid level in the chamber 303 and the direction of the centrifugal force are taken as a reference. The liquid level only reaches the level. Therefore, if the flow path 304 is lifted above the upper side surface of the lower chamber in the same manner as the flow path 302 described above, the sample liquid moves to the front of the next chamber while the centrifugal force is acting. Never do.
When the centrifugal force is removed by stopping the rotation or the like, the sample solution immediately moves through the flow path 304 by capillary action and reaches the next chamber, in FIG. 5, the trouble of the transmitted light measurement chamber 305. To do. By applying a centrifugal force from this state, the sample liquid flows into the transmitted light measurement chamber 305. If the action of the centrifugal force is stopped in this state, the sample liquid in the transmitted light measurement chamber 305 may flow backward through the channel 304 due to capillary action, and the amount of sample liquid in the transmitted light measurement chamber may be insufficient. For this reason, a centrifugal force is applied also when measuring transmitted light.

In order to facilitate the inflow of the sample solution between the chambers, air holes 306, 307, and 308 are provided in the portion where the sample solution cannot reach at the upper part of each chamber. With such a structure, it is possible to sufficiently dissolve and react with the sample solution and move the flow path.
When the reaction reagent necessary for measuring the specific component in the sample solution is dried and supported in the chamber 302, an aqueous solution having a reagent concentration higher than the concentration necessary for the reaction is dripped and dried, or the volume of the chamber 302 is reduced. When the reaction reagent is dissolved in an amount of the sample solution, the reaction reagent layer is formed by dripping and drying the reagent solution at a concentration and a drop amount so that a sufficient amount of the reagent necessary for the reaction can be supported in the chamber 302. Obtainable.

However, in the configuration in which the reagent layer is formed in the chamber 302, when the sample liquid flows in, the effect of stirring cannot be obtained, and thus the reagent layer may not be sufficiently dissolved.
In addition, when the connection portion between the bottom surface and the side surface of the chamber 302 is a right angle or an acute angle, the dried reagent layer tends to be deposited more heavily on this portion than in the vicinity of the center of the bottom surface. In some cases, the reagent layer was not sufficiently dissolved during the inflow of the liquid. In either case, there is a concern that the reagent concentration in the sample solution is not sufficient, resulting in a problem that the chemical reaction necessary for measurement does not proceed sufficiently.

Furthermore, when the reagent layer is obtained by air-drying the reagent solution, the surface of the reagent layer becomes dense, which is a factor that hinders dissolution by the sample solution. In addition, in the air drying process of the reagent solution, the reagent solution is very concentrated due to evaporation of moisture, so that there is a possibility that the reagent solution may be denatured depending on the composition of the reagent layer.
International Publication No. 00/026667 Pamphlet JP 2000-580007 A

  The present invention relates to an analytical disc having means for detecting the chemical reaction of a mixed solution of the sample and the reagent, which solves the above-mentioned conventional problems, and in particular, makes the reaction of the sample and the reagent quick and accurate. It aims at ensuring the accuracy of the chemical change detection.

In order to solve the above problems, the present invention provides:
A chamber composed of a gap provided between the upper substrate and the lower substrate via a spacer, and at least two openings provided in the chamber;
The chamber includes a plurality of lower concave portions provided on the lower substrate, a solid reagent containing a substance necessary for a reaction with a liquid analysis sample and dissolved in the liquid analysis sample, characterized Oite at at least one portion in the lower concave portion, and the lower recessed portion, the said lower concave portion facing the portion of the substrate, that, that the solid reagent is gripped A sample analysis disk is provided.

In the sample analysis disk, the chamber includes a plurality of upper concave portions provided on the upper substrate corresponding to the lower concave portions,
It Oite at least one portion in the portion which is formed by the combination of the upper concave portion and the lower concave portion, said solid reagent by said lower concave portion and the upper concave portion is grasped Are preferred.

In addition, the solid reagent is obtained by freezing an aqueous solution of the substance and then heating it under reduced pressure to sublimate water, or formed by compression molding the powder of the substance. It is preferable that

According to the present invention, in the analytical disc for analyzing the sample by detecting a chemical reaction between the sample and the solid reagent supported in the chamber, the solid reagent is denatured by containing moisture. can be prevented, also in the surface and inside of the solid reagents to improve the solubility of the reagent layer by voids sample liquid penetrates is formed, enables shortening of the reaction time, out chemical reactions detection allowing ensure the accuracy and speed of detection of a specific component in the sample liquid by. Furthermore, since the solid reagent can be produced independently of the analytical disk, an improvement in flexibility in the production process can be expected.

Hereinafter, embodiments of the present invention will be described with reference to the drawings.
FIG. 1 is a schematic top view showing an embodiment of a chamber included in the sample analysis disk of the present invention. In FIG. 1, the sample analysis disk is indicated by a one-dot chain line, and the center of rotation is indicated by C. 2 is a cross-sectional view taken along the line aa of the chamber 10 portion of the sample analysis disk shown in FIG.
As shown in FIGS. 1 and 2, the chamber 10 of the sample analysis disk according to the present embodiment is composed of a gap provided between the upper substrate 6 and the lower substrate 8 via a spacer 7, and includes at least two pieces. Openings 1 and 2 are provided. The chamber 10 includes a plurality of lower concave portions 3 a provided on the lower substrate 6, and a solid reagent is carried in at least one of the lower concave portions 3 a.

Chamber 10 is configured with an upper substrate 6 and the lower substrate 8 may be mounted on a disk that is composed of separate members, also the chamber 10 is constituted by an upper substrate 6 and the lower substrate 8 In addition, the disk itself may be composed of the upper substrate 6 and the lower substrate 8. Further, the chamber 10 constituted by the upper substrate 6 and the lower substrate 8 may be built in the disk. However, in FIG. 1 and 2, the chamber 10 is composed of an upper substrate 6 and the lower substrate 8, the disk is shown in phantom at one point chain line in FIG. 1.

The chamber 10 is formed by bonding the upper substrate 6, the spacer 7, and the lower substrate 8, and the lower substrate 8 is provided with a curved concave portion 3 a that forms a part of the side surface and the bottom surface of the chamber 10. The spacer 7 is provided with a cutout portion corresponding to a side wall of the flow path and a part of the side wall of the chamber 10.
The upper substrate 6 forms a flow path and a ceiling surface of the chamber 10. Curved concave portions 3a and 3b are provided at equal intervals on the bottom surface of the chamber 10 and the ceiling surface at a position opposite thereto.

The concave portions 3a and 3b are configured to have a curved surface corresponding to a part of the spherical surface.
In the chamber 10 of the present invention, is the diameter of a sphere virtually constructed from the curved surface of the concave portion 3a and the corresponding curved surface of the concave portion 3b equal to the distance between the deepest portions of the concave portions 3a and 3b? It is preferable that the upper substrate 6 and the lower substrate 8 are provided with concave portions 3a and 3b, respectively, so as to be larger. This is because the solid reagent 5 can be held by the concave portions 3a and 3b.
At least one of the concave portions 3a and 3b is sandwiched with a solid reagent 5 molded into a substantially spherical shape (including a spherical shape).

More specifically, the diameter of the solid reagent 5, concave portions 3a, at a depth D ₁ less the chamber 10 in 3b, and the upper substrate 6 on the bottom side the lower substrate 8 and the ceiling surface side of the chamber 10, It is preferable that the depth D ₂ or more in a plane region without the concave portions 3a and 3b. This is because, by satisfying this condition, the solid reagent 5 can be physically and reliably fixed in the chamber 10.
The concave portions 3a and 3b are respectively formed by the number of the solid reagents 5, but the distance between the centers of the concave portions 3a and 3b is preferably equal to or larger than the diameter of the solid reagent 5.

The solid reagent 5 is a solidified reagent necessary for the reaction. The reagent powder is sealed in a spherical mold having the diameter of the desired solid reagent 5 and compressed, or droplets of an aqueous solution of the reagent are added. It can be obtained by freezing and drying by controlling the diameter of the desired solid reagent 5. Thus the solid reagent 5 manufactured by a method, easily dissolved in a sample which has flowed into the chamber 10, it is easy to ensure a concentration required for the reaction.
However, the method is limited to the above method as long as the solid reagent 5 that can be packaged in the chamber 10 in an amount sufficient to ensure the concentration necessary for the reaction when dissolved with the sample flowing into the chamber 10 can be obtained. is not.

The solid reagent 5 may be a single substance or a mixture of a plurality of substances. Moreover, the solid reagent 5 actually packaged in one chamber 10 is not necessarily one type, and may be a plurality of types.
For example, if 10 kinds of substances are originally required for a chemical reaction for detecting a specific component in a sample, if one solid reagent 5 that originally contains all 10 kinds of substances is used, It is convenient that the chemical reaction proceeds immediately after the solid reagent 5 is dissolved in the sample. However, among the 10 types of substances, when a reagent with mixed repellency that interferes with each other by contact and gradually denatures is included, it is preferable to use one solid reagent 5 that includes all substances. Therefore, if the 10 kinds of substances are divided into two groups, for example, and two kinds of solid reagents 5 containing each substance group are packaged in one chamber 10, the mixed repellent substances can be separated, and During the reaction, they can be mixed at once.

A plurality of one kind of solid reagent 5 can be packaged in one chamber 10. In this case, the diameter of the solid reagent 5 can be reduced, and the depth of the chamber 10 and the thickness of the disk in the concave portions 3a and 3b can be reduced.
Moreover, the surface area per mass of the spherical surface of the solid reagent 5 is increased, and it can be expected that dissolution of the sample proceeds more rapidly when the sample enters the chamber 10.

The sample flows in from the flow path connected to the opening 1 near the disk rotation center C, and flows out from the opening 2 farthest from the disk rotation center C.
Furthermore, even when the concave portion 3a or the concave portion 3b is formed on either the bottom surface or the ceiling portion of the chamber 10, the above-described effects can be obtained. In this case, the physical fixing effect of the solid reagent 5 is slightly weaker than when there are concave portions above and below, but the solid reagent 5 is on the inner wall of the chamber 10 (the ceiling surface or the bottom surface facing the concave portion). The area in contact with the sample becomes smaller, and the solubility of the sample is improved accordingly.

The sample flows into the chamber 10 having the structure described above from the flow path connected to the opening 1 close to the disk rotation center C, dissolves the solid reagent 5 in the chamber 10, and the opening farthest from the disk rotation center C is opened. It flows out from the part 2 and is transferred to the next chamber (not shown).
In the following, the present invention is described in more detail by way of examples. In addition, in order to describe the present invention more specifically, specific measurement objects, materials of members, and the like are specifically described, but the present invention is not limited by those descriptions.

In order to measure the total cholesterol concentration in plasma, a chamber 10 mounted on a liquid sample analysis disk having the structure shown in FIGS. 1 and 2 was prepared.
An upper substrate 6 and a lower substrate 8 made of polycarbonate and a spacer 7 made of polyethylene terephthalate and coated with an adhesive on both sides were prepared. When the lower substrate 8 was produced by molding, the concave portion 3a was provided. Further, when the upper substrate 6 was produced by molding, the concave portion 3b was provided. The concave portions 3a and 3b were provided so as to face each other when the upper substrate 6, the spacer 7 and the lower substrate 8 were bonded together.

When the direction in which centrifugal force is applied during the rotation of the disk (the direction of the arrow in FIG. 1) is “vertical direction”, the portion of the upper substrate 6 and the lower substrate 8 facing the chamber 10 is the normal direction of the surface of the disk. In view of the above, it was configured to have a rectangular planar shape with a vertical _Dt of 1.5 mm and a horizontal _Dw of 4 mm.
More specifically, among the rectangles, 0.5 mm, 1.5 mm, 2.5 mm from the left side of the rectangle at a position t (= 0.5 mm) from the side on the outer side when the disk rotates. Four curved concave portions 3a and 3b each having a diameter of 0.8 mm and a depth of 0.2 mm were provided on the upper substrate 6 and the lower substrate 8 at positions of 3.5 mm and 3.5 mm, respectively.

Further, the portion where the lower substrate 8 faces the chamber 10 was given a depth of 0.5 mm. Therefore, when the upper substrate 6, the spacer 7 having a thickness of 100 μm, and the lower substrate 8 are bonded together, the distance from the flat surface portion of the bottom surface of the chamber 10 to the flat surface portion of the ceiling, that is, the depth of the chamber 10 is 0.6 mm.
In this case, the concave portions 3a and 3b form a curved surface shape and a positional relationship that form the surface of a virtual sphere having a diameter of 1.0 mm. The volume of the chamber 10 was about 4.03 μl including the volume of the concave portions 3a and 3b, but as will be described later, the solid reagent 5 having a diameter of 1.0 mm at the position of the concave portions 3a and 3b. If the sample flows in approximately 2.83 μl or more, the solid reagent 5 can be completely immersed in the sample. Therefore, the disk chamber 10 and the flow path were configured to control the amount of sample flowing into the chamber 10 to 3.0 μl.

As will be described later, the solid reagent 5 is formed by lyophilizing a solution in which a group of reagents necessary for the reaction is dissolved in water or a pH buffer solution. However, in the reagent system for detecting cholesterol, all the reagents are 1 When two layers are used, the expected reaction is unlikely to occur due to denaturation due to the interaction between reagents.
Therefore, the solid reagent 5 divided into three types was arranged in two chambers as described later. Further, in the conventional reagent analysis disk shown in FIG. 5, the chamber 10 described above is used as the chambers 301, 303 and 305. Note that it is not necessary to dispose the solid reagent 5 in the transmitted light measurement chamber 305, the upstream chamber 301 in FIG. 5 corresponds to the first chamber, and the downstream chamber 303 is the second chamber. It corresponds to.

The sample transfer mechanism is the same as the sample solution transfer mechanism of the conventional sample analysis disk.
The 1st chamber 301 and the 2nd chamber 303 are connected by the flow path of the form mentioned above. Subsequent to the two chambers 301 and 303, a measurement chamber 305 for detecting a chemical change in the sample is connected.
In this example, the chemical change was detected by detecting the absorbance change at a specific wavelength of the dye contained in the solid reagent 5 due to the chemical reaction of cholesterol. Therefore, the bottom surface and the ceiling of the measurement chamber were flat and optically transparent to the wavelength.

The depth of the measurement chamber is appropriately 200 μm when using a reagent described later as a reagent, but generally corresponds to the optical path length during transmitted light measurement, so that the amount of transmitted light or absorbance changes during measurement. Therefore, the transmitted light amount or absorbance of the substance indicating the concentration of the substance to be measured may be appropriately set so as to be an appropriate numerical value.
The chambers 301 and 303 for containing the solid reagent 5 have a volume of about 4 μl as described above, and the solid reagent 5 is completely dissolved if the amount of sample flowing in is controlled to 3 μl. On the other hand, the measurement chamber 305 has a planar shape with a diameter of 2 mm and a depth of 200 μm.
In this case, since the volume of the measurement chamber 305 is about 0.6 μl, it is possible to secure the amount of sample liquid necessary for measurement even if a sample remains in the flow channel during the flow channel transfer.

The following reaction mechanism is used to measure the plasma cholesterol concentration.
(1) E-Chol → Chol (enzyme: ChE)
(2) Chol + NAD → cholestenone + NADH (enzyme: ChDH)
(3) NADH + WST-9 → NAD + formazan (enzyme: DI)
The total cholesterol concentration is calculated by measuring the amount of change corresponding to the concentration of the absorbance with respect to transmitted light having a wavelength of 650 nm due to the formation of formazan in the above formula (3). In the above formulas (1) to (3), E-Chol represents a cholesterol ester. Most of the cholesterol in plasma is esterified. Chol indicates cholesterol. ChE indicates cholesterol esterase (EC 3.1.1.13) which is an enzyme that catalyzes a reaction for changing E-Chol to Chol.

  ChDH is cholesterol dehydrogenase (EC is unknown, purchased from Amano Enzyme Co., Ltd.), NAD is nicotine adenine dinucleotide which is a coenzyme of ChDH, and NADH indicates the reduced state of NAD. WST-9 is an abbreviation for “water-soluble tetrazolium-9” which is an acronym for English, and is a kind of tetrazolium salt available from Dojindo Laboratories. DI represents diaphorase (EC 1.6.99.2). DI is an enzyme that catalyzes the oxidation reaction of NADH to NAD and the reduction reaction coupled thereto.

In order to perform the above reaction, the following three kinds of solid reagents 5 are separately arranged in the chambers 301 and 303. The first chamber 301 contains ChE, an enzyme solid containing ChDH, and a NAD solid containing NAD and a surfactant, and the second chamber 303 contains a WST solid containing WST-9 and DI. include.
Furthermore, since the suitable pH of ChDH is 8 or more (alkaline region), it is preferable to use a pH buffer in the reaction system. However, Tris hydrochloride, which is a pH buffer for adjusting the pH during the reaction, is freeze-dried and immediately after the lyophilized solid is exposed to normal air in a mixed solution with an enzyme or a surfactant. There is a tendency to absorb and deliquesce water. Therefore, a mixed solution with sodium cholate, which is one of the surfactants added for the purpose of activating the reaction system, was dropped onto the concave portion 3b of the first chamber 301 and dried.

The enzyme solid consists of ChE, ChDH, and sucrose added to maintain the solid shape after lyophilization. As long as the additive for maintaining the shape does not affect the reaction system, sugars other than sucrose, proteins, and the like can be used as appropriate.
The NAD solid is composed of n-octyl-β-D-thioglucoside and NAD, which are surfactants for activating the catalytic activity of ChE. In particular, the shape can be maintained without adding any other additives, but additives can also be added to make it more rigid.
The WST solid consists of WST-9, DI and sucrose. As described above, sucrose is added to maintain the shape.

From the viewpoint of reactivity and measurement time, it is preferable that the number of solid reagents 5 be as small as possible. However, in the case of the reaction system in this example, the mixed aqueous solution of ChDH and NAD is not stable. So these two materials need to be separate solids. Therefore, in order to separate these two reagents, a solid reagent 5 containing an enzyme and a solid reagent containing NAD were prepared.
These two kinds of solid reagents 5 were placed in one chamber because the oxidation reaction of cholesterol in the sample and the reduction reaction of NAD coupled thereto proceeded only after both were dissolved in the sample solution.

Furthermore, since WST-9 has a tendency to inhibit the catalytic activity of ChDH, a solid reagent 5 containing WST-9 was also prepared independently, and this was caused by the progress of the reaction with the two solid reagents. Since it was necessary to dissolve in the sample after that, it was necessary to arrange in a separate chamber 303.
Along with WST-9, DI was also added to the same reagent solid to catalyze the reduction reaction of WST-9.

Since the reactions of the above formulas (1) and (2) are performed under conditions that do not contain WST-9, and then WST-9 is mixed to induce a color reaction, the sample flows in the order of flow. A solid reagent containing an enzyme and a solid reagent containing NAD were placed in the first chamber 301, and a solid reagent containing WST-9 was placed in the second chamber 303.
Each solid reagent is formed by lyophilization so as to have a diameter of 1 mm. Two solid reagents containing an enzyme and two solid reagents containing NAD are each in the first chamber 301, and each solid reagent containing WST is Four pieces were arranged in the second chamber 303.

  The concentration of the aqueous reagent solution for obtaining each solid reagent is adjusted to a concentration required for the reaction with 3 μl of the sample. That is, the volume of 1 mm in diameter is about 0.52 μl, and two solid reagents containing an enzyme and two solid reagents containing NAD are arranged, so that each reagent aqueous solution has a concentration required for the reaction in the sample. The solid reagent containing WST was adjusted to 3 / (2 × 0.52) times and 3 / (4 × 0.52) times. These density adjustments need to be appropriately determined in consideration of the shapes of the chambers 301 and 303.

The standard serum or a sample diluted with physiological saline was spotted on a liquid sample analysis disk containing the solid reagent 5 prepared by the method described above, and the absorbance was measured with a measuring apparatus having the structure shown in FIG. However, a change in absorbance (that is, responsiveness) depending on the total cholesterol concentration in the sample solution could be observed. The result is shown in FIG.
In FIG. 6, the vertical axis indicates the absorbance when laser light with a wavelength of 650 nm is irradiated, and the horizontal axis indicates the cholesterol concentration (free type conversion) in the sample solution. From this result, it can be seen that the change in absorbance depends on the cholesterol concentration.

  In addition, although the present Example showed the example which measures the total cholesterol density | concentration in plasma by detecting the light absorbency change of WST-9 which is a pigment | dye, when it became an aqueous solution instead of WST-9, Felicia An electrode capable of containing at least a counter electrode and a working electrode may be provided in the measurement chamber by containing potassium ferricyanide that generates halide ions in a solid reagent.

In this case, the ferrocyanide ion generated by reducing ferricyanide ions by oxidation of cholesterol in plasma is further provided in the analyzer of FIG. 3 by providing a terminal that can contact the electrode from the outside of the disk. It is also possible to measure and measure an oxidation current value generated when re-oxidizing by applying a voltage between the electrodes. In this case, a redox compound that can exchange electrons with NADH can be arbitrarily used instead of potassium ferricyanide.
Furthermore, in addition to the cholesterol in the plasma shown in this example, it is possible to establish a reaction system capable of optically or electrochemically detecting changes caused by a chemical reaction with a target component. The present invention can be used for any measurement object.

  When analyzing a sample by detecting a chemical change in a reagent that has reacted with a sample, the liquid sample analysis disk according to the present invention dissolves the reagent quickly and uniformly with a smaller amount of sample, and the chemical reaction is performed. It is possible to proceed without spatial bias, so that it is possible to ensure the accuracy of detection of a specific component in a sample solution by chemical reaction detection, which is useful for a blood component measuring device or the like.

1 is a schematic top view of an embodiment of a sample analysis disk of the present invention. FIG. It is the sectional view on the aa line of the sample analysis disk shown in FIG. It is a figure which shows the structure of the disk for analysis used with the conventional liquid sample analyzer. It is a figure which shows the structure of the conventional analyzer. It is a schematic diagram for demonstrating the mechanism of the sample transfer in the conventional sample analysis disc and the sample analysis disc of this invention. It is a graph which shows the responsiveness in an Example.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1, 2 Opening part 3a, 3b Concave part 5 Solid reagent 6 Upper board | substrate 7 Spacer 8 Lower board | substrate 101 Liquid sample analysis disk 104 Sample inlet 105 Flow path 106 Reagent 201 Spindle motor 900 Sample 212 Optical pickup 213 Feed motor 302, 304 Flow path 301 Upstream chamber 303 Downstream chamber 305 Transmitted light measurement chamber 306, 307, 308 Air port

Claims (9)

A chamber composed of a gap provided between the upper substrate and the lower substrate via a spacer, and at least two openings provided in the chamber;
The chamber includes a plurality of lower concave portions provided on the lower substrate , and a solid reagent containing a substance necessary for reaction with the liquid analysis sample and dissolved in the liquid analysis sample ,
Oite at at least one portion in the lower concave portion, and the lower recessed portion, the said lower concave portion facing the portion of the substrate, that, that the solid reagent is gripped Characteristic sample analysis disc. The chamber includes a plurality of upper concave portions provided on the upper substrate corresponding to the lower concave portions;
It Oite at least one portion in the portion which is formed by the combination of the upper concave portion and the lower concave portion, said solid reagent by said lower concave portion and the upper concave portion is grasped The sample analysis disk according to claim 1, wherein:   3. The sample analysis disk according to claim 1, wherein the lower concave portion has a curved surface constituting a part of a spherical surface.   The sample analysis disk according to claim 2, wherein the upper concave portion has a curved surface constituting a part of a spherical surface.   The sample analysis disk according to any one of claims 1 to 4, wherein the solid reagent is solidified in a substantially spherical shape.   The disk for sample analysis according to claim 1, wherein the solid reagent includes a single substance or a mixture of a plurality of substances.   The sample analysis disk according to any one of claims 1 to 6, wherein the substance includes a plurality of mixed repellent substances, and the plurality of mixed repellent substances are separately solidified. The solid reagent, after freezing the aqueous solution of the materials are heated under reduced pressure, it is obtained by sublimating the moisture, in any one of claims 1-7, characterized in specimen worth析用disk described. The solid reagent, specimen partial析用disk according to any one of claims 1 to 8, the powder of the material that is one formed by compression molding, characterized by.

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