JP2004144739A - Sampling element for specimen, specimen treatment apparatus, and specimen treatment method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a technique which simplifies the whole pre-treatment step, in inspection of a specimen, especially including specimen collection and specimen treatment which are often performed manually, introduction of a specimen to a measurement (reaction) system and specimen dilution, and improves the operability of the inspection using a large-sized automated device and a POCT-corresponding device. <P>SOLUTION: Provided is a sampling element for specimen having a first region which can quantitatively collect and temporarily hold a specimen, and a second region which is subjected to dynamic actions from the exterior in order to make the first region move. <P>COPYRIGHT: (C)2004,JPO

Description

{Circle over (2)} The present invention relates to an element for simply processing the sample when analyzing a substance contained in the sample, and a sample processing method using the element. Therefore, the present invention mainly belongs to the field of clinical testing.

In recent years, with the progress of analysis technology, analysis technology, and inspection technology, it has become possible to measure various substances. Particularly in the field of clinical testing, the development of a measurement principle based on a specific reaction such as a biochemical reaction, an enzymatic reaction, or an immune reaction has enabled the measurement of substances in body fluids that are reflected in a disease state. Following this, various large-scale automation devices have been developed for the purpose of measuring more items for more samples (multi-sample multi-item measurement). According to such an automatic apparatus, high sensitivity and high precision measurement can be performed by improving the performance of the components constituting the apparatus and the performance of the reaction reagent used in the apparatus.

Currently, the central clinical laboratory of a large hospital such as a general hospital owns a large-scale automated device, where a wide variety of samples sent daily are tested. In general, a sample to be measured by an automated apparatus is subjected to a pretreatment, and if necessary, a dilution operation or the like is performed, and thereafter, the measurement is started. For example, in the case of blood, a pretreatment for separating blood cells and plasma is performed. Therefore, the user of the test equipment at the central clinical laboratory requires the minimum required expertise.

On the other hand, in recent years, a field of clinical testing called point-of-care testing (POCT) has attracted attention. The first object of the POCT is to shorten the time from the collection of a specimen to the time when a test result is obtained, based on a concept contradicting the examination performed at the above-mentioned central clinical laboratory, and to provide a simple and quick measurement as a first object. Therefore, the POCT requires a simple measurement principle and a small, highly portable and operable measurement device. 2. Description of the Related Art Recent advances in technological development have led to the development of small measuring devices that can be easily measured, for example, as represented by blood glucose sensors.

In addition, POCT has the following effects: in addition to enabling quick and accurate diagnosis by obtaining quick measurement results, the cost of the test is reduced, and the burden on the subject due to the reduction in the amount of blood and other samples is reduced. Reduction and reduction of infectious waste can be considered. At present, clinical examinations are rapidly shifting to POCT, and measuring instruments compatible with POCT are being developed to meet the needs.

As a measuring device corresponding to POCT, there is a qualitative immunosensor using an immunoantigen-antibody reaction typified by a pregnancy diagnostic drug, in addition to the enzyme sensor using an enzyme electrode reaction typified by the blood glucose sensor described above. Recently, quantitative immunosensors have also been developed and commercialized. All are based on the construction of the simple measurement principle and the accompanying advancement of the solid-phase technology of the biological component, the technology of the sensor device, and the technology of the sensor system. Further, the technology for downsizing a large-sized automation device owned by the above-mentioned central clinical laboratory is also being developed.

By the way, the inspection of a sample generally includes (1) sample collection, (2) sample processing, (3) sample introduction into a measurement (reaction) system, (4) sample dilution, (5) reagent introduction, (6) Reaction, (7) unreacted matter removal, (8) detection, and (9) measurement of a substance in a sample are performed in nine steps. Further, in the large-scale automation device, the steps (3) to (9) are mainly automated, and a multi-sample multi-item measurement is possible.

However, the steps (1) and (2), for example, steps for separating blood cells from blood and removing contaminants are not automated. Therefore, these steps occupy a large part of the inspection time and are an obstacle to obtaining quick results. Furthermore, it requires expertise in sample processing, and is not an operation that anyone can perform. In addition, some of the dilution operations required for a sample containing a high-concentration protein are partially automated, but on the other hand, the configuration of the apparatus is complicated, and the cost is high for use in POCT. . Further, in the future, even if a large-scale automation device is downsized by a technology such as μ-TAS in consideration of POCT, which is an important position in clinical examination, it is expected that a problem will occur in the sample pretreatment process.

On the other hand, for example, a POCT-compatible device such as a blood glucose sensor does not require the step (2), and the steps (3) to (8) are performed in a short time in the process in which the sample flows into the sensor device after the step (1). Done in The same applies to pregnancy diagnostics. However, not all POCT-compatible devices can take such a sensor system, and again, manual operations are required in the steps (1) and (2), and sometimes (3) and (4). There were many problems. In other words, most of the current POCT-compatible devices simply perform only the steps (4) to (9) (for example, Patent Document 1).
JP-A-6-160388

In view of the above prior art, the present invention can be applied to any automated small-sized device or POCT-compatible device, and can easily perform the steps (1) to (3), (4) or (5) with good operability. It is an object of the present invention to provide a sample sampling element that can be used, and a sample processing apparatus and a sample processing method using the same.

In order to achieve the above object, the present invention provides a first region capable of quantitatively collecting a sample and temporarily holding the sample, and receiving a mechanical action from outside to move the first region. A sample sampling element having a second region is provided.

It is effective that the first region holds the specimen by capillary action.
It is also effective that the mechanical action is caused by a change in a magnetic field.
In addition, it is effective that the first region releases the specimen due to the movement of the second region.

The specimen sampling device, it is effective to retain the reagent b ₁ to disrupt the cells contained in the reagent a ₁ and / or said analyte to react with the substances contained in the specimen.
The reagent a _1, the enzyme is effective antigen, antibody, be a receptor or nucleic acid.

Further, it is effective that the substance is a protein, a hormone, an antibody, an enzyme, an antigen or a nucleic acid.
The reagent b _1, it is effective that an inorganic salt or a surfactant.

Further, it is effective that the cells are red blood cells, white blood cells or platelets.
Components released from the cells destroyed by the reagent b ₁ is a protein, glycated protein, it is effective that a phosphorylated protein, hormones, lipids, antibodies, enzymes, antigens, receptors, inhibitors, DNA or RNA .

Further, the present invention provides a sample having a first region capable of quantitatively collecting a sample and temporarily holding the sample, and a second region which is subjected to a mechanical action from the outside in order to move the first region. A sampling element, a reaction cell into which the sampling element is introduced, means for applying a mechanical action to the sampling element in the reaction cell, and an optical measurement system for measuring a reaction in the reaction cell. A sample processing apparatus is provided.

In this sample processing apparatus, it is effective that the means for giving a mechanical action is a magnetic field changing device for giving a mechanical action to the sampling element by a magnetic force.
Further, it is effective that the optical measurement system is a light scattering spectrophotometer, a fluorescence spectrophotometer, an absorption spectrophotometer or an emission spectrophotometer.

Further, the present invention provides (a) a first region capable of quantitatively collecting a sample and temporarily holding the same, and a second region which is subjected to a mechanical action from the outside to move the first region. (B) introducing the sampling element holding the sample into a reaction system; and (c) introducing the sampling element holding the sample into the reaction system. Moving the sampling element by mechanical action, releasing the sample from the sampling element, and uniformly mixing the sample with the reaction system while stirring. Also provide.

Prior to the step (a), the it is effective to carry the reagent b ₁ to disrupt the cells contained in the reagent a ₁ and / or said analyte to react with the substances contained in the sample to the sampling device.
Also in this sample processing method, the reagent a ₁ is, it is effective is an enzyme, antigen, antibody, receptor or nucleic acid.

Further, it is effective that the substance is a protein, a hormone, an antibody, an enzyme, an antigen or a nucleic acid.
The reagent b _1, it is effective that an inorganic salt or a surfactant.
Advantageously, said cells are red blood cells, white blood cells or platelets.
Components released from the cells destroyed by the reagent b ₁ are proteins, glycosylated proteins, it is effective that a phosphorylated protein, hormones, lipids, antibodies, enzymes, antigens, receptors, inhibitors, DNA or RNA .

Further, the reaction system, buffer, it is effective to include a solution containing reagents b ₂ to disrupt the cells contained in the solution or the sample, comprising a reagent a ₂ which reacts with the substance contained in the sample .
In the step (c), the reagent a ₂ reacts with a substance contained in the sample, and / or the reagent b ₂ simultaneously mixes the sample and the reaction system uniformly while stirring. It is effective to destroy the cells contained in.

ADVANTAGE OF THE INVENTION According to the sampling element which concerns on this invention, and the sample processing apparatus and the sample processing method using the same, the sampling of the sample in analysis, analysis, and a test and the pre-processing can be performed easily, and also the simple and quick measurement can be performed. This can contribute to the development of POCT-compatible devices.

The sample sampling element according to the present invention includes a first region capable of quantitatively collecting a sample and temporarily holding the sample, and a second region that is subjected to a mechanical action from the outside to move the first region. It has a region. In addition, it is effective that the first region can hold the specimen by capillary action, and the mechanical action is caused by a change in a magnetic field.

Therefore, specific examples of the sample sampling element according to the present invention include, for example, a capillary capable of quantitatively collecting and holding a sample, and a first magnetic body provided in contact with the capillary. . That is, in this case, the first region corresponds to a capillary, and the second region corresponds to a first magnetic body. The sample sampling element according to the present invention includes a capillary and a magnetic body (material). The feature is that it is composed.

If this sample sampling element is used, as will be described later, for example, by moving the second magnetic body by a mechanical action using a magnetic field from the outside, the capillary is moved together with the first magnetic body, and centrifugal force is generated. The sample can be released from the capillary by, for example, and mixed with the liquid system outside the capillary while stirring.

Here, a capillary is also referred to as a capillary or a capillary, and is obtained by laminating a thin tube having a thickness enough to cause a capillary phenomenon, or laminating two or more flat plates or the like with a gap enough to cause a capillary phenomenon. Refers to a complex obtained. For example, a yarn obtained by twisting thin fibers is used as a warp and a weft, and a cloth or the like obtained by weaving these yarns can be made into a capillary by providing a gap between fibers or between yarns to the extent that a capillary phenomenon appears. Can be used as In any case, those having a capillary phenomenon are collectively called capillary tubes.

Next, the capillary phenomenon refers to a phenomenon in which after a solid and a liquid reach an equilibrium state at an interface, they have a certain shape or a certain area, the surface tension of the liquid, the wettability of the solid to the liquid, And three factors of capillary diameter are involved. The relationship between these three factors and the capillary phenomenon will be briefly described by taking a phenomenon in which water rises in a glass tube as an example. Intermolecular force (cohesion) acts between the water molecules, but since the water molecules on the water surface are in contact with air, they are pulled only in the direction toward the water. This force acting on the surface of the water is called surface tension, which is why the water surface is generally round.

If a capillary such as a thin glass tube is vertically attached to water in such a state, the force acting between the glass tube and the water molecule (adhesive force and wettability) is more than the intermolecular force (cohesion force) acting on the water molecule. Is larger, the water is drawn to the wall of the glass tube and rises, and the water moves upward to a position where the adhesion and the weight of the raised water are balanced. At this time, as the glass tube is thinner, the weight of the water to be raised is lighter, so that the water moves to a higher position to balance the adhesive force. Therefore, the material constituting the capillary may be any material having hydrophilicity.

尺度 The measure of wettability can be easily obtained by, for example, dropping water on a flat plate or the like made of the same material as the material constituting the capillary, and examining the extent of the spread. That is, it can be said that the larger the spread, the easier the material is to get wet. Specifically, glass, quartz, iron or the like can be used, and any of them can constitute a capillary tube. The function required for the capillary in the present invention is to quantitatively collect, introduce, and hold a sample in the capillary. That is, sampling is performed in a series of operations until various kinds of analysis of the sample are performed.

On the other hand, a magnetic substance is a substance having a property of magnetizing in a magnetic field (magnetism) and interacting with the magnetic field of the substance itself. All substances react in response to a magnetic field and are classified into a ferromagnetic substance strongly attracted in the direction of the magnetic field, a paramagnetic substance weakly attracted, and a diamagnetic substance exhibiting weak repulsion. The magnetic material in the present invention particularly refers to a ferromagnetic material or a paramagnetic material, for example, a metal material such as iron attracted to a magnet. Here, the first magnetic body and the second magnetic body in the present invention have a property of attracting each other.

Particularly, the function required for the first magnetic body is to perform some kind of movement according to the movement of the second magnetic body in addition to being attracted to the second magnetic body such as a magnet. For example, when the second magnetic body makes a rotational movement, the first magnetic body also makes a rotational movement, and when the second magnetic body makes an amplitude movement, the first magnetic body also makes an amplitude movement, or If the second magnetic body makes a rotational movement, the first magnetic body needs to be able to make a simple oscillation. Therefore, it is necessary that the first magnetic body has a weight and a size that can be moved by the second magnetic body.

サ ン プ リ ン グ In the sampling element according to the present invention, the capillary and the first magnetic body are integrally formed. Therefore, the capillary itself also moves by the movement of the first magnetic body. As a method of integrating both, for example, a method of winding a capillary tube and a first magnetic body with an adhesive material such as a tape, a method of winding a capillary tube and a first magnetic body with a film made of a fluororesin such as Teflon, or the like. Examples include a method of wrapping with a material, and a method of forming a capillary tube itself with a magnetic material. In any case, it is important that the capillary is integrated so that the capillary can move together with the first magnetic body without detaching from the first magnetic body by the movement of the first magnetic body. It is not done.

Here, FIG. 1 is a schematic perspective view of a sampling element according to the present invention. As shown in FIG. 1, the sampling element 1 according to the present invention is configured by integrating a capillary 11 and a first magnetic body 12 with, for example, an adhesive tape 13. For reference, FIG. 2 shows a sampling element viewed from the direction of arrow X in FIG. 1, and FIG. 3 shows a sampling element viewed from the direction of arrow Y in FIG.

Furthermore, the sampling element according to the present invention thus configured may carry the first reagent. As a form of loading, for example, a method in which a second capillary is further integrated with the first magnetic substance, and a first reagent in a solution state or a dried state is loaded on the second capillary, A method in which a container-like part is formed and the first reagent is supported in a dry state in the part, or a carrier such as a glass fiber filter paper or a cellulosic filter paper impregnated and supported with the first reagent is sampled. A method of integrating the element with the element may be used. In any case, it is necessary that the first reagent is supported so as not to spill, and that the area where the first reagent is supported is not in a sealed state and is exposed to the outside.

Here, FIG. 4 is a schematic perspective view of another sampling element according to the present invention. As shown in FIG. 4, the sampling element 1 according to the present invention is configured by integrating a capillary 21 and a first magnetic body 22 with, for example, an adhesive tape 24. Further, a glass fiber filter paper 23 impregnated with and carrying the first reagent is integrally held between the capillary 21 and the first magnetic body 22. For reference, FIG. 5 shows a sampling element viewed from the direction of arrow X in FIG. 4, and FIG. 6 shows a sampling element viewed from the direction of arrow Y in FIG.

The first reagent to be supported, include a reagent a ₁ which reacts with the substance contained in the sample, for example, enzymes, antigens, antibodies, etc. receptor or nucleic acid.
Examples of the enzyme include glucose oxidase, which specifically reacts with glucose in a sample.
Examples of the antigen include a protein that specifically reacts with an antibody mainly present in a specimen. Note that the antibody present in the sample includes, for example, an HCV antibody or an HIV antibody.

Examples of the antibody include a polyclonal antibody, a monoclonal antibody, a chimeric antibody, a Fab antibody, an F (ab) 2 antibody, and an Fv antibody. The antibody can be prepared from an antigen present in the sample. The antigen present in the sample is a protein, glycated protein, phosphorylated protein, hapten, or the like.
Examples of the receptor include those which specifically react with a physiologically active substance such as steroid, thyroid hormone, vitamin, peptide hormone, growth factor, cytokine, or catecholamine contained in the sample.
Examples of the nucleic acid include those that can be genes such as DNA or RNA.

Therefore, the substance contained in the specimen is a protein, a hormone, an antibody, an enzyme, an antigen or a nucleic acid.
Examples of the protein include a protein composed of at least one of 20 kinds of amino acids such as albumin, globulin, myoglobin, prolamin, glutelin, histone and protamine, a C-reactive protein (CRP) of an infectious disease marker, and serum amyloid A ( SAA), tumor marker carcinoembryonic antigen (CEA), α-fetoprotein (AFP), carbohydrate antigen 19-9 (CA19-9), glycated hemoglobin (HbA1c), glycoalbumin, and other saccharide-protein complexes , All phosphorylated proteins obtained by phosphoric acid ester bond to the hydroxyl groups of serine, threonine and tyrosine residues of proteins, and high-density lipoprotein (HDL) and low-density lipoprotein (LDL) ) And all lipoproteins that are complexes of lipids and proteins. Such as quality and the like.

Hormones include human chorionic hormone (hCG), luteinizing hormone (LH), triiodothyronine (T3), thyroxine (T4), thyroid stimulating hormone (TSH), insulin and the like.
Examples of the enzyme include glutamate oxaloacetate transaminase (GOT), alkaline phosphatase (ALP), glutamate pyruvate transaminase (GPT), lactate dehydrogenase (LDH), leucine aminopeptidase (LAP), γ-glutamyl transpeptidase ( γ-GTP), cholinesterase (ChE), acid phosphatase (ACP), and prostate acid phosphatase (PAP).
Antibodies include HCV antibodies, HIV antibodies and immunoglobulins (IgE), and antigens include viruses and bacteria.
Examples of the nucleic acid include SNPS, DNA and RNA.

The first reagent to be supported is, it may be a reagent b ₁ to disrupt the cells contained in the sample. Such reagents b _1, for example, inorganic salts or surfactants. Inorganic salts change the osmotic pressure inside and outside the cell, and promote cell destruction by contracting or expanding the cell. On the other hand, surfactants promote cell destruction by disrupting the balance between hydrophilicity and hydrophobicity of proteins as cell components.
Such inorganic salts include, for example, sodium chloride, potassium chloride, sodium fluoride, sodium thiocyanate, potassium thiocyanate, and the like, and surfactants include, for example, sucrose monolaurate, sodium oleate, and dodecyl sulfate Sodium (SDS) and the like.

As the cells are destroyed by the reagent b _1, for example, red blood cells, white blood cells, and platelets, and the like. In this case, an object is to mix or dissolve substances inside red blood cells, white blood cells or platelets in an extracellular liquid system.
Components released from the disrupted cells include, for example, proteins, glycated proteins, phosphorylated proteins, hormones, lipids, antibodies, enzymes, antigens, receptors, inhibitors, DNA, RNA, and the like.

Further, the sampling device according to the present invention, the reagent b ₁ to disrupt the cells contained in the reagent a ₁ and analyte which reacts with the substance contained in the specimen may be supported simultaneously. In this case, although the reagent b ₁ to disrupt the cells contained in the reagent a ₁ and analyte which reacts with the substance contained in the specimen shall not affect each other, these combinations, the effect of the present invention Can be appropriately selected by a person skilled in the art within a range that does not impair.

Here, the purpose of using the sampling element according to the present invention is to easily and quickly sample a sample and to easily and quickly discharge the sample into a reaction system (liquid system). Therefore, the present invention also relates to a sample processing apparatus and a sample processing method using the sampling element. The sample processing method referred to here includes mixing, dissolving, and diluting a sample in a predetermined liquid system.

The sample processing apparatus according to the present invention, the sample sampling element, a reaction cell into which the sampling element is introduced, a means for applying a mechanical action to the sampling element in the reaction cell, a reaction in the reaction cell And an optical measurement system for measuring

The means for giving a mechanical action may be any magnetic field changing device that gives a mechanical action to the sampling element by a magnetic force. For example, a magnetic stirrer described later can be used.
Further, a light scattering spectrophotometer, a fluorescence spectrophotometer, an absorption spectrophotometer or an emission spectrophotometer may be used for the optical measurement system.

The sample processing method using the sample sampling element according to the present invention can be performed by the sample processing apparatus. Specifically, (a) a sample is quantitatively collected and temporarily stored. A step of causing a sample sampling element having a first region to be obtained and a second region which is subjected to a mechanical action from the outside to move the first region to quantitatively collect and hold a sample; b) a step of introducing the sampling element holding the sample into a reaction system, and (c) moving the sampling element by a mechanical action from outside the reaction system to release the sample from the sampling element. And uniformly mixing the sample and the reaction system while stirring.

Prior to the step (a), the be supported reagents b ₁ to disrupt the cells contained in the reagent a ₁ and / or said analyte to react with the substances contained in the sample to the sampling device is preferred.
Further, in the step (c) and, at the same time homogeneously mixed with stirring and the reaction system and the analyte is reacted with the substance contained the reagent a ₂ in the specimen, and / or by the reagent b ₂ The cells contained in the specimen can be destroyed.

Here, a sample according to the present invention is typified by using a sampling element obtained by integrating a glass capillary tube (glass tube) with a stirring bar (stirrer bar) used for organic synthesis reaction and solid dissolution. The processing method will be described.
First, a specimen is quantitatively raised and held (sampled) in a glass tube, and the sampling element is brought into contact with (preferably immersed in) a target liquid system (reaction system).

Next, using a device called a magnetic stirrer, for example, a stirring bar (first magnetic body) in the liquid system is rotated from outside the liquid system. The specimen held in the glass tube is released into the liquid system by this rotational movement, and the specimen and the liquid are uniformly mixed or dissolved. The stirrer includes a second magnetic body.
Since the glass tube and the stirring bar are integrated, the glass tube also integrally rotates by rotating the stirring bar. Therefore, the specimen held in the glass tube is released into the liquid by centrifugal force, and is further uniformly mixed by stirring.

Note that the shape of the sampling element is not limited to a rod shape, and the sampling element can move in the liquid with the movement of the second magnetic substance (for example, a magnet) outside the liquid, and the sample is released and uniformly mixed and stirred with the whole liquid. It only needs to have a certain weight and size. In addition, the movement of the second magnetic body from outside the liquid system is not limited to the rotational movement of the stirrer as described above, and may be, for example, an amplitude movement or a simple vibration. What is important is that the liquid efficiently discharges the sample held by the sampling element, and furthermore, exercises a motion of uniformly mixing with the liquid.

Therefore, the sampling element according to the present invention has three important functions: a “sample sampling function”, a “release function of releasing a sample into a liquid”, and a “uniformization function of uniformly mixing a sample with a liquid”. Sampling refers to quantitatively collecting and holding a sample in a capillary tube using capillary action, and releasing refers to releasing the sample held in the capillary tube into a liquid. Further, homogenization means that the sample and the liquid are uniformly mixed, and the substance contained in the sample is present at a constant concentration in all parts of the obtained mixture.

Further, the sample processing method according to the present invention includes a step of quantitatively injecting the sample into the capillary of the sampling element by capillary action, and a step of moving the sampling element into which the sample is injected by a magnetic force from outside the liquid system in a liquid. In addition, it can be said that the method includes a step of uniformly mixing the sample in the capillary tube into the liquid during the exercise.
By using the sample processing method of the present invention, substances contained in the sample can be easily diluted. For example, hemoglobin present at a high concentration in red blood cells in blood can be easily diluted by the above method. Furthermore, if the liquid contains a reagent that reacts with the substance in the specimen, a reaction for analyzing the substance in the specimen can be performed immediately after the homogeneous dissolution of the specimen.

Here, the liquid and the used in the sample processing method according to the present invention, a buffer, a solution containing a reagent a ₂ which reacts with the substance contained in the specimen, or a reagent b ₂ to disrupt the cells contained in the sample Containing solution.
Examples of the buffer include a phosphate buffer, a Tris buffer, and a carbonate buffer.
As the reagent a ₂ which reacts with the substance contained in the specimen, it is possible to use those listed as a reagent a ₁ which reacts with the substance contained in the specimen as described above, to destroy the cells contained in the sample reagents b _2, can be used those listed as a reagent b ₁ to disrupt the cells contained in the specimen as described above.

When the sampling element according to the present invention further supports the first reagent, the first reagent may be released into the liquid during the movement of the sampling element. At this time, the first reagent released is a reagent a ₁ which reacts with a substance in the sample, there is a case where the substance in the sample and the reagent a ₁ is at the same time the reaction in a homogeneous mixing process.
In addition, examples of the sample applied to the sample homogeneous mixing method of the present invention include body fluids, and more specifically, for example, blood, urine, saliva, tears, and sweat.
The reactions that can take place in the sample homogeneous mixing process according to the present invention include, for example, an enzyme reaction, an immune reaction, a hybridization, a hemolysis reaction, and a lysis reaction.

Here, a uniform mixing method using the sample sampling element and the stirrer according to the present invention will be described with reference to the drawings.
FIG. 7 is a process chart of a sample processing method using a sampling element according to the present invention. Here, the case where the sampling element 1 shown in FIG. 1 is used will be described, but the adhesive tape 13 is omitted. First, as shown in FIG. 7A, the sampling element 1 is brought into contact with the sample 30, and the sample 30 is introduced into the capillary 11 by capillary action. Thereby, the sample 30 is held in the capillary 11 of the sampling element 1 as shown in FIG.

Next, the sampling element 1 is introduced into a reaction cell 31 (reaction system) containing a liquid 33, and a magnet 32, which is a second magnetic material constituting a stirrer, is positioned outside the bottom surface of the reaction cell 31. I do. As the second magnetic body 32 rotates, the sampling element 1 rotates in the liquid, and the sample held in the capillary 11 is released into the liquid 33 by centrifugal force (FIG. 7 (c)). . Then, both are uniformly mixed by the stirring by the rotational movement of the sampling element 1. The sample 30 contained in the capillary tube 11 of the sampling element 1 is mixed with the liquid 33, and at this time, a mixed liquid (or lysis liquid) 34 exists in the reaction cell 31 ((d) in FIG. 7). )).

Next, in order to carry out the sample processing method using the sampling element, the present invention also provides a handling device for the sample sampling element. This handling device is for a sample having a first region capable of quantitatively collecting a sample and temporarily holding the sample, and a second region which is subjected to a mechanical action from the outside to move the first region. A sampling element is detachably included, and the sampling element holding the sample can be introduced into a reaction system.

Further, it is effective that the handling device has a mechanism capable of holding a liquid system constituting the reaction system and introducing the liquid system into the reaction system together with the sampling element.
The liquid system, buffer solution may be a solution containing reagents b ₂ to disrupt the cells contained in the solution or the sample, comprising a reagent a ₂ which reacts with the substance contained in the specimen.

Here, FIG. 8 shows a process chart of a sample processing method using the sampling element and the handling device according to the present invention. As shown in FIG. 8A, a handling device 40 according to the present invention includes a syringe 43 having a piston 43a and holding a liquid 43b, and a syringe 43 for holding the liquid 43b inside the syringe 43. It includes an aluminum seal 44 provided on the bottom. The sampling element 1 according to the present invention is attached to the tip of the syringe 43.

When this handling device 40 is used, the specimen 30 is held in advance in the capillary tube 11 of the sampling element 1 as shown in FIG. 8A, and then, as shown in FIG. By pushing the piston 43 a of 43, the liquid 43 b is introduced into the reaction cell 46 together with the sampling element 1. Thereafter, as shown in FIG. 8 (c), the sampling element 1 holding the sample 30 and the reaction cell 46 containing the liquid 43b are placed on, for example, a stirrer, and the sampling element 1 is rotated to release the sample 30. And uniformly mix with the liquid 43b while stirring.

Here, the features of the handling device 40 shown in FIG. 8 are that it has a syringe shape, that the liquid 43b to be mixed with the sample is sealed, that the detachable sampling element 1 is provided at the tip, and that the piston 43a is pressed. In addition, the sampling element 1 comes off, and the aluminum seal 44 is broken, so that the liquid 43b flows out. The syringe-shaped part is preferably made of plastic, for example, and is preferably disposable.
Further, the aluminum seal 44 has a function of holding the liquid 43b in a state where it does not leak and easily breaking the liquid 43b by an external force. Therefore, in place of the aluminum seal 44, a rupturable material having such a function may be used.

Next, FIG. 9 shows a state in which the sampling element 1 is attached and detached from the distal end portion of the handling device 40 shown in FIG. FIG. 9 is an enlarged view of a distal end portion of the handling device 40.
As shown in FIG. 9, the member 53 includes a core 53b and a cylinder 53a that can move up and down around the core 53b. In this case, the first magnetic body 51 of the sampling element 50 has a cylindrical shape and is held in a state of being pierced by the core 53b. When the cylinder 53a is moved downward, the held sampling element 50 is pushed by the cylinder 53a and comes off the core 53b.

8 and 9, the functions of the handling device 40 are summarized. First, the sample 30 is introduced into the capillary 11 of the sampling device 1 as shown in FIG. 8 using the handling device 40 provided with the sampling device 1. Next, by pushing the piston 43a of the syringe 43 toward the inside of the reaction cell 46, the sampling element 1 is introduced into the reaction cell 46 by a detachable function as shown in FIG. And the liquid 43b is also introduced into the reaction cell 46. Thereafter, operations as shown in FIGS. 7C and 7D are performed.

Further, a sample processing method using another handling device according to the present invention will be described. As shown in FIG. 10A, the handling device 60 is configured such that a reaction cell 63a holding a liquid 63b is sealed with a sampling element 1. A sample is held in the capillary 11 of the sampling element 1 (not shown).
In addition, the handling device 60 includes a lid 62 having a mechanism capable of dropping the sampling element 1 into the reaction cell 63a in the direction of arrow Z as shown in FIG. By pushing the lid 62 vertically downward, the sampling element 1 is introduced into the reaction cell 63a. 8 and 10, sampling, liquid introduction, and uniform mixing can be easily performed.

Next, an embodiment of a sample processing apparatus according to the present invention using the above-described sample sampling element will be described with reference to FIGS. FIG. 11 is a diagram showing a configuration in which a part of the sample processing apparatus according to the present invention is formed in a cross section. The sample processing apparatus measures a reaction in a reaction cell 70 into which the sampling element is introduced, a means (for example, a magnetic stirrer) 71 for applying a mechanical action to the sampling element in the reaction cell 70, and a reaction in the reaction cell 70. And an optical measurement system. The optical measurement system includes a light source 72 and a photometer (detector) 73, and a sampling element (not shown) according to the present invention is introduced into the reaction cell 70.

The reaction cell 70 is made of, for example, transparent quartz, and the liquid system in the reaction cell 70 is irradiated with substantially parallel light (or laser) 74 emitted from the light source 72. For example, first, a predetermined reagent (liquid system) is put in the reaction cell 70, and then, a sampling element that includes a capillary (first region) and a magnetic substance (second region) and holds a sample in the capillary. Is introduced into the reaction cell 70. Thereafter, the sampling element subjected to the mechanical action from the means 71 for applying a mechanical action rotates, and the specimen is discharged from the capillary (first region) into the reaction cell 70.

Due to the reaction between the sample and the reagent, for example, the liquid system in the reaction cell 70 becomes cloudy, and the intensity of the scattered light 76 before and after the reaction of the liquid system is measured by the photometer 73, so that the reaction between the sample and the reagent Completion or quantification or qualification of the sample can be performed, and the sample can be suitably processed. The sample processing apparatus may be controlled by the control device 75.
Hereinafter, a sample processing method using a sampling element according to the present invention will be described in detail with reference to examples, but the present invention is not limited thereto.

<< Example 1 >>
In this example, first, the sampling element according to the present invention shown in FIG. 1 was manufactured. A glass tube (EM Meister Minicaps manufactured by AS ONE Corporation) was used as the capillary tube 11, and a stirrer bar (Laboran rotator manufactured by AS ONE Corporation) was used as the first magnetic body 12. The sampling element according to the present invention having the structure shown in FIG. 1 was produced by winding the glass tube and the stirrer bar with the adhesive tape 13 and integrating them. The capacity of the glass tube was 5 ul, and the size of the stirrer bar was 25 mm × φ8 mm.

<< Examples 2 to 6 and Comparative Example 1 >>
In the present example, the sample processing method according to the present invention was performed according to the procedure shown in FIG. 7 using the sampling element manufactured in Example 1, and the effect of dilution of the sample was examined.
First, a sampling element according to the present invention was manufactured in the same manner as in Example 1. In order to observe the manner in which a sample is mixed and dissolved in a liquid by an absorption spectrum, 20 mg of a dye (BRILLIANT GREEN manufactured by Wako Pure Chemical Industries, Ltd.) is dissolved in 1 ml of distilled water, and a sample (BG) having a concentration of 20 mg / ml is dissolved. Solution).

After injecting the sample into a 5 ul glass tube of the sampling element, the sampling element was placed in a beaker containing 5 ml of distilled water, and immediately stirred with a stirrer (MIGHTY MAGNETIC STIRRER MODEL-16 manufactured by KOIKE PRECISION INSTRUMENT). . After stirring for 1 minute, 1 ml of the liquid in the beaker was sampled and its absorption spectrum was measured. The results are shown in FIG. 13A (Example 2).
On the other hand, as before, 5 ul of the same specimen as in Example 2 was collected with a pipetteman (manufactured by Gilson), placed in a 5 ml aqueous solution being stirred by a stirrer in a beaker, and stirred for 1 minute. Thereafter, 1 ml of the liquid in the beaker was sampled and its absorption spectrum was measured. The result was shown by b of FIG. 13 (Comparative Example 1).
From FIG. 13, it was found that even by using the sample processing method using the sampling element according to the present invention, uniform mixing of the samples can be performed as before, and further, a simple sample processing method can be provided.

サ ン プ リ ン グ Furthermore, a sampling device was prepared in the same manner as in Example 1, and the dilution effect of the sample was analyzed. Using a 1 μl, 3 μl, 5 μl or 10 ul glass tube (EM Meister Minicaps manufactured by AS ONE Corporation) and a stirrer bar (25 mm × 8 mm), the sampling element 4 having a volume of 1 μl, 3 μl, 5 μl or 10 ul was used. Seeds were made. The same sample as in Example 2 was injected into four types of sampling elements, each was introduced into a 5 ml aqueous solution contained in a beaker, and diluted by stirring with a stirrer. Thereafter, 1 ml of the liquid in the beaker was sampled and its absorption spectrum was measured. The results are shown in FIG. 14 (Examples 3 to 6). From FIG. 14, it was possible to prepare a dilution series of the sample by using the sample processing method using the sampling element according to the present invention.

<< Examples 7-11 >>
In this example, a sampling element having the structure shown in FIG. 4 and supporting potassium chloride was prepared, and hemolysis of blood was performed.
A 0.1%, 0.5%, 1%, 5% or 10% potassium chloride solution impregnated and freeze-dried glass fiber filter paper 23 (Whatman F075-14, 10 mm × 30 mm) is prepared. The sample is placed between a glass tube having a capacity of 3 ul, which is a capillary tube 21, and a stirrer bar, which is a first magnetic body 22, wrapped with an adhesive tape 24, and five kinds of sampling elements carrying potassium chloride. Produced.
These were put into 1 ml of whole blood, stirred for 3 minutes, and then blood cells were separated by centrifugation. The globin concentration in the plasma thus obtained was measured using an SLS-Hb measurement kit (manufactured by Wako Pure Chemical Industries, Ltd.). The results are shown in FIG. From FIG. 15, it can be seen that the possibility of controlling the hemolysis of blood was shown by using a sampling element carrying potassium chloride.

<< Example 12 >>
In this example, human serum albumin measurement was performed using the sampling element having the structure shown in FIG. 1 according to the present invention.
A sampling element including a glass tube having a capacity of 3 ul as a capillary tube was produced in the same manner as in Example 1. In addition, the concentration of the specimen is 0 g / l, 0.005 g / l, 0.015 g / l, 0.046 g / l, 0.23 g / l, 0.46 g / l, 1.5 g / l or 4.6 g. / L human serum albumin was used.
The measurement was performed by injecting 3 ul of human serum albumin into the sampling element, and then injecting the sampling element into a 1 ml buffer solution (50 mM MOPS, 4% PEG, pH 7.4) containing a rabbit-derived anti-human albumin antibody and a mouse-derived anti-human albumin antibody. After stirring for 3 minutes, the scattering intensity (measurement wavelength 670 nm) was measured. The results are shown in FIG. As shown in FIG. 16, it was confirmed that human serum albumin can be measured using the sampling device according to the present invention. This indicated the possibility of being applicable to measurement items other than human serum albumin.

検 体 The sample sampling element, sample processing device and sample processing method according to the present invention can be suitably used for, for example, urinalysis, pregnancy test, blood test, immune serum test, endocrine function test, virus test, bacterial test and the like.

1 is a schematic perspective view of a sampling device according to the present invention. FIG. 2 is a diagram showing a sampling element viewed from a direction of an arrow X in FIG. 1. FIG. 2 is a diagram illustrating a sampling element viewed from a direction of an arrow Y in FIG. 1. FIG. 7 is a schematic perspective view of another sampling element according to the present invention. FIG. 5 is a diagram illustrating a sampling element viewed from a direction of an arrow X in FIG. 4. FIG. 5 is a diagram illustrating a sampling element viewed from a direction of an arrow Y in FIG. 4. It is a figure showing a process of a sample processing method using a sampling element concerning the present invention. FIG. 4 is a diagram showing steps of a sample processing method using the sample processing apparatus according to the present invention. FIG. 9 is a diagram illustrating a state in which a sampling element 50 is attached and detached from a tip portion of the handling device 40 illustrated in FIG. 8. It is a figure showing a process of a sample processing method using another handling device concerning the present invention. FIG. 1 is a schematic side view illustrating a configuration of a sample processing apparatus according to the present invention. FIG. 12 is a schematic top view of the sample processing apparatus shown in FIG. 11. It is a graph which shows the absorption spectrum in the Example of this invention, and the absorption spectrum in a comparative example. It is a graph which shows the dilution series in the example of the present invention. It is a graph which shows the measurement result of the hemolysis reaction in the example of the present invention. It is a graph which shows the measurement result of human serum albumin in the example of the present invention.

Explanation of reference numerals

DESCRIPTION OF SYMBOLS 1 Sampling element 11, 21 Capillary tube 12, 22 First magnetic body 13, 24 Adhesive tape 23 Glass fiber filter paper 30 Sample 31, 63a Reaction cell 32 Second magnetic body 33, 63b Liquid 34 Mixed liquid 40, 60 Handling device 43 Syringe 43a Piston 43b Liquid 44 Aluminum Seal 46 Reaction Cell 50 Sampling Element 51 First Magnetic Body 52 Capillary Tube 53 Member 62 Lid 70 Reaction Cell 71 Means for Applying Mechanical Action 72 Light Source 73 Photometer 74 Nearly Parallel Light 75 Controller

Claims (22)

A sample for quantitatively collecting a sample and temporarily holding the sample, and a second region which is subjected to a mechanical action from the outside to move the first region. Sampling element. 2. The sample sampling element according to claim 1, wherein the first area holds the sample by capillary action. 2. The sample sampling element according to claim 1, wherein the mechanical action is caused by a change in a magnetic field. 2. The sample sampling element according to claim 1, wherein the first region emits the sample by the movement of the second region. Specimen sampling device of claim 1, wherein the holding the reagent b ₁ to disrupt the cells contained in the reagent a ₁ and / or said analyte to react with the substances contained in the specimen. The reagent a ₁ is, enzymes, antigens, antibodies, analyte sampling device according to claim 5, characterized in that the receptor or nucleic acid. The sample sampling element according to claim 5, wherein the substance is a protein, hormone, antibody, enzyme, antigen or nucleic acid. The reagent b ₁ is analyte sampling device according to claim 5, wherein the inorganic salt or a surfactant. The sample sampling element according to claim 5, wherein the cells are red blood cells, white blood cells, or platelets. Components released from the cells destroyed by the reagent b ₁ is, for proteins, glycosylated proteins, phosphorylated proteins, hormones, lipids, wherein antibodies, enzymes, antigens, receptors, inhibitors, that it is a DNA or RNA A sample sampling element according to claim 5. A sample sampling element having a first region capable of quantitatively collecting a sample and temporarily holding the sample, and a second region that is subjected to a mechanical action from the outside to move the first region;
A reaction cell into which the sampling element is introduced,
Means for applying a mechanical action to the sampling element in the reaction cell;
A sample processing apparatus, comprising: an optical measurement system for measuring a reaction in the reaction cell. 12. The sample processing apparatus according to claim 11, wherein the means for giving a mechanical action is a magnetic field changing device for giving a mechanical action to the sampling element by a magnetic force. 12. The sample processing apparatus according to claim 11, wherein the optical measurement system is a light scattering spectrophotometer, a fluorescence spectrophotometer, an absorption spectrophotometer, or an emission spectrophotometer. (A) a sample sampling element having a first region capable of quantitatively collecting a sample and temporarily holding the sample, and a second region which is subjected to a mechanical action from the outside to move the first region A step of quantitatively collecting and holding the sample,
(B) introducing the sampling element holding the sample into a reaction system;
(C) moving the sampling element by mechanical action from outside the reaction system to release the sample from the sampling element, and mixing the sample into the reaction system while stirring. Characteristic sample processing method. Prior to the step (a), the claims, characterized in that for supporting the reagent b ₁ to disrupt the cells contained in the reagent a ₁ and / or said analyte to react with the substances contained in the sample to the sampling device 15. The sample processing method according to 14. Sample processing method of claim 15, wherein said reagent a ₁ is, enzymes, antigens, antibodies, receptors, or nucleic acids. 16. The sample processing method according to claim 15, wherein the substance is a protein, a hormone, an antibody, an enzyme, an antigen, or a nucleic acid. The reagent b ₁ is, sample processing method of claim 15, wherein the inorganic salt or a surfactant. The method according to claim 15, wherein the cells are red blood cells, white blood cells, or platelets. Components released from the cells destroyed by the reagent b ₁ is, for proteins, glycosylated proteins, phosphorylated proteins, hormones, lipids, wherein antibodies, enzymes, antigens, receptors, inhibitors, that it is a DNA or RNA The sample processing method according to claim 15. The reaction system, buffers, claim 14, characterized in that a solution containing reagents b ₂ to disrupt the cells contained in the solution or the sample, comprising a reagent a ₂ which reacts with the substance contained in the sample The sample processing method according to the above. In the step (c), the reagent a ₂ reacts with a substance contained in the sample, and / or the reagent b ₂ is contained in the sample, while mixing the sample into the reaction system while stirring. The sample processing method according to claim 14, wherein cells are destroyed.

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