WO2019181400A1 - Specimen processing method and vessel for specimen processing - Google Patents

Abstract

The present invention addresses the problem of providing a specimen processing method and a vessel for specimen processing that make reliable detection using magnetic force and fluorescence possible even if there is a large amount of a target substance within a specimen. To solve this problem, in the present invention, a specimen is added to a liquid in a vessel, a conjugate of a target substance in the specimen and magnetic particles is formed in the liquid, magnetic force is used to separate the conjugate and the liquid containing the specimen within the vessel, and the liquid containing the specimen is immobilized within the vessel after the conjugate and the liquid containing the specimen are separated.

Description

Sample processing method and sample processing container

The present invention relates to a sample processing method and a sample processing container used for detecting a target substance such as bacteria from a sample.

In detection of micro-organisms such as proteins, viruses and bacteria, it is preferable that detection is possible with high sensitivity even if the target substance is a small amount. In particular, it is important that viruses such as influenza virus and norovirus, which are concerned about the spread of infectious diseases, can be reliably detected even if the amount in the sample is very small.

As a highly sensitive detection method capable of detecting a small amount of virus or the like, a polymerase chain reaction (PCR) is known.
In the polymerase chain reaction method, by amplifying only the base sequence to be examined by polymerase chain reaction, the virus to be examined can be amplified about 1 million times in 20 cycles. Therefore, it is possible to detect highly sensitive viruses and the like.
On the other hand, the polymerase chain reaction method has problems such as the influence of impurities is large, the pretreatment is complicated, and refrigeration or frozen storage such as a reagent test is necessary.

On the other hand, as a method for easily detecting a small amount of virus or the like, a conjugate described in Patent Document 1 that forms a near field on the detection plate and contains a target substance (target substance) on the detection plate surface There is known an optical detection method for detecting an optical signal as an optical signal.
Specifically, the optical detection method described in Patent Document 1 generates a conjugate in which fluorescent particles (phosphors) and magnetic particles are bound to a target substance using, for example, an antigen-antibody reaction. On the other hand, a near field is generated on the surface of the detection plate by light irradiated from the back side under total reflection conditions. In this state, the generated combined body is attracted to the near field (detection plate) by magnetic force and is moved in parallel to the detection plate. In the optical detection method described in Patent Document 1, the target substance is detected by measuring the light amount fluctuation caused by the movement of the subject and the movement of light (bright spot).

International Publication No. 2017/187744

In the optical detection method that binds the fluorescent particles to the target substance, the non-specific adsorption causes the fluorescent particles adsorbed to other than the target substance to become noise, the detection sensitivity decreases, and the highly sensitive target substance cannot be detected. There is a problem.
On the other hand, in the method described in Patent Document 1, the non-specifically adsorbed fluorescent particles other than the target substance do not move by magnetic force, and only the conjugate of the magnetic particles, the target substance and the fluorescent particles is fluorescent. It moves by magnetic force while generating. Therefore, according to the method described in Patent Document 1, by detecting the movement of light (bright spot), noise due to nonspecifically adsorbed fluorescent particles can be removed and the target substance can be detected with high sensitivity. .

However, in the method described in Patent Document 1, when the amount of the target substance present in the subject is large, the target substance may not be detected even though the target substance exists in the subject. .
That is, when the amount of the target substance in the subject is very large compared to the amount of the magnetic particles and the fluorescent particles, the combination of the target substance and the magnetic particles without the magnetic particles and the fluorescent particles, In addition, there is a possibility that only a conjugate of the target substance and fluorescent particles is generated, and a conjugate in which the target substance is bonded with magnetic particles and fluorescent particles is not generated.

The conjugate of the target substance and magnetic particles moves by magnetic force but does not emit light. On the other hand, the conjugate of the target substance and the fluorescent particles emits light but does not move by magnetic force.
That is, in this case, there is no conjugate of the target substance, magnetic particles, and fluorescent particles that moves by magnetic force while generating light despite the presence of the target substance in the subject. The substance cannot be detected, resulting in false detection that there is no target substance in the subject.
In the detection of viruses that are likely to spread infectious diseases such as influenza viruses and noroviruses, false detection of the absence of a virus is a major problem even though the target substance virus is present in the specimen. .

An object of the present invention is to solve such problems of the prior art, and in the detection of a target substance from a subject using magnetic force and fluorescence, even when the amount of the target substance in the subject is large. Another object of the present invention is to provide a specimen processing method that makes it possible to properly detect a target substance, and a specimen processing container used in the specimen processing method.

In order to solve this problem, the present invention has the following configuration.
[1] A specimen is supplied into a liquid in a container to form a combined body in which a target substance in the specimen and magnetic particles are bound in the liquid,
In the container, the conjugate and the liquid containing the analyte are separated by magnetic force,
A method for processing a specimen, comprising: separating a conjugate and a liquid containing a specimen, and then immobilizing the liquid containing the specimen in a container.
[2] The specimen processing method according to [1], wherein after immobilizing a liquid containing the specimen, the fluorescent particles are bound to the conjugate.
[3] The specimen processing method according to [2], wherein the binding of the conjugate and the fluorescent particles is performed in a space in which a fixed body in which a liquid containing the specimen is immobilized is part of the wall surface.
[4] The subject processing method according to [3], wherein the immobilized body is a bottom surface of the space.
[5] The method according to any one of [2] to [4], wherein the binding of the conjugate and the fluorescent particles is performed by supplying the liquid containing the fluorescent particles to the conjugate separated from the liquid containing the analyte. Analyte processing method.
[6] Liquid containing fluorescent particles is stored in a supply chamber having the wall surface of the container as a part of its wall surface, and the liquid containing fluorescent particles is supplied to the conjugate separated from the liquid containing the analyte. The method for processing a subject according to [5], wherein the method is performed by breaking a part of the supply chamber.
[7] The subject processing method according to [6], wherein the container is tubular, and the supply chamber has a double-tube structure in which the wall of the container is one tube.
[8] The subject processing method according to [6] or [7], wherein the inner wall surface of the container is a part of the wall surface of the supply chamber.
[9] The subject processing method according to any one of [1] to [8], wherein the container is separated into a plurality of chambers by walls that can be broken by a subject collection tool.
[10] The subject processing method according to [9], wherein the substance is moved between the plurality of chambers by gravity.
[11] One of the plurality of chambers is a liquid chamber for storing a liquid, and another one of the plurality of chambers is a magnetic particle chamber for storing magnetic particles, and among the plurality of chambers The subject processing method according to [9] or [10], wherein the another chamber is an immobilization chamber for immobilizing a liquid containing the subject.
[12] In the container, the liquid chamber, the magnetic particle chamber, and the immobilization chamber are provided in one direction in the order of the liquid chamber, the magnetic particle chamber, and the immobilization chamber. Method.
[13] The subject processing method according to [11] or [12], wherein the liquid chamber and the magnetic particle chamber, and the magnetic particle chamber and the immobilization chamber are adjacent to each other.
[14] The subject processing method according to any one of [11] to [13], wherein the immobilization chamber has at least one of a disinfectant and a disinfectant.
[15] The method for processing a specimen according to any one of [1] to [14], wherein the container has a blocking agent.
[16] A container for processing a subject,
The interior is separated into a plurality of chambers by a wall that can be broken by a specimen collection tool, and one of the plurality of chambers contains a liquid to be supplied with the subject, An object processing container, wherein another chamber contains magnetic particles, and another one of the plurality of chambers contains a substance for immobilizing a liquid.
[17] The specimen processing container according to [16], which has a chamber for storing fluorescent particles in at least one of the inside of the container and the outside of the container.

According to the present invention, in the detection of a target substance from a subject using magnetic force and fluorescence, the target substance can be detected properly even when the amount of the target substance in the subject is large.

FIG. 1 is a diagram conceptually illustrating an example of a subject processing container according to the present invention, which implements an example of a subject processing method according to the present invention. FIG. 2 is a conceptual diagram for explaining an example of the object processing method of the present invention. FIG. 3 is a conceptual diagram for explaining an example of the subject processing method of the present invention. FIG. 4 is a diagram conceptually illustrating an example of a detection apparatus that detects a target substance using the subject processing container of the present invention. FIG. 5 is a top view of the detection apparatus shown in FIG. FIG. 6 is a conceptual diagram for explaining the operation of the detection device shown in FIG. FIG. 7 is a diagram conceptually illustrating another example of the subject processing container of the present invention, which implements another example of the subject processing method of the present invention. FIG. 8 is a diagram conceptually illustrating another example of the subject processing container of the present invention, which implements another example of the subject processing method of the present invention.

Hereinafter, a subject processing method and a subject processing container according to the present invention will be described in detail based on preferred embodiments shown in the accompanying drawings.

In the present specification, a numerical range expressed using “to” means a range including numerical values described before and after “to” as a lower limit value and an upper limit value.

FIG. 1 shows a conceptual diagram of an example of a subject processing container according to the present invention, which implements an example of a subject processing method according to the present invention.

In the subject processing method of the present invention, first, the subject is supplied to the liquid in the container to form a conjugate in which the target substance in the subject and the magnetic particles are bound in the liquid. Next, in the container, the conjugate and the liquid containing the analyte are separated using magnetic force. Thereafter, the liquid containing the analyte separated from the conjugate is immobilized in the container.
In the specimen treatment method of the present invention, preferably, after the liquid containing the specimen is immobilized in the container, the fluorescent particles are further bound to the conjugate obtained by binding the target substance and the magnetic particles. Thus, a combined body in which the target substance, magnetic particles, and fluorescent particles are combined is formed.

A subject processing container 10 shown in FIG. 1 is for carrying out such a subject processing method of the present invention, and has a rectangular cylindrical container main body 12 having one end opened and the other end closed. The container body 12 includes a liquid chamber 16, a magnetic particle chamber 18, and an immobilization chamber 20 that are separated by the partition wall 14 a and the partition wall 14 b and are arranged in the height direction of the square cylinder.
Furthermore, as a preferable aspect, the specimen processing container 10 has a double tube structure in which a rectangular cylindrical inner tube 24 having both surfaces opened is inserted into a region corresponding to the liquid chamber 16 of the container body 12. Yes. The end surface on the magnetic particle chamber 18 side between the container body 12 and the inner tube 24 is closed by a partition wall 14c, and the space between the container body 12 and the inner tube 24 becomes the fluorescent particle supply chamber 28. Yes. Accordingly, the liquid chamber 16 is inside the inner tube 24. The fluorescent particle supply chamber 28 is provided with a breaking jig 30.
Both the partition wall 14a and the partition wall 14b are films that can be broken by a sample collection tool such as a swab. The partition wall 14 c forming the fluorescent particle supply chamber 28 is a film that can be broken by the breaking jig 30.

In such a specimen processing container 10, the liquid chamber 16 contains a liquid L1 for dissolving and / or dispersing the specimen.
Magnetic particles are accommodated in the magnetic particle chamber 18.
In the immobilization chamber 20, a substance for immobilizing the liquid L1 accommodated in the liquid chamber 16 is accommodated.
Further, the fluorescent particle supply chamber 28 contains a liquid L2 containing fluorescent particles.

Thus, the specimen processing container 10 contains a liquid, and is normally used with the closed end side of the container body 12 facing downward.
In the following description, the closed end side of the container body 12 is also referred to as the bottom. Further, the end surface on the closed end side of the container body 12 is also referred to as a bottom surface.

In the present invention, the target substance (target substance) to be detected is not limited. Examples include viruses, bacteria, DNA (deoxyribonucleic acid), RNA (ribonucleic acid), proteins, and contaminants.
Moreover, there is no restriction | limiting also in the test object (sample to collect) used as the detection object of a target substance, The various things considered to contain the target substance can be utilized. Examples of the subject include body fluids such as blood and lymph, saliva, sweat, runny nose, tears, vomit, urine, feces, chemicals, environmental water, clean water, sewage, and wipes.
These specimens may be collected by a known method according to the specimen. For example, a method of collecting a subject by wiping a doorknob, a table, or the like at a site where food poisoning or the like has occurred with a swab (cotton swab) or the like is exemplified. As another method, a method of collecting a subject by bringing a swab into contact with vomit, urine and the like is exemplified.

There are no limitations on magnetic particles (magnetic particles, magnetic materials, magnetic substances), and various known particles used for detection of target substances using magnetism can be used. Examples include magnetic beads and magnetic powder. Commercial products are also available for these.
There is no restriction on the amount of saturation magnetization of the magnetic particles. Saturation magnetization of the magnetic particles is preferably ^{0.01 ~ 200A · m 2 / kg} , more preferably ^{0.1 ~ 100A · m 2 / kg} , more preferably ^{0.3 ~ 50A · m 2 / kg} .
By setting the saturation magnetization amount of the magnetic particles to 0.01 A · m ² / kg or more, the combined body of the magnetic particles, the target substance, and the fluorescent particles can be suitably moved by applying a magnetic force described later. By setting the saturation magnetization amount of the magnetic particles to 100 A · m ² / kg or less, the combined body reaches the inner wall of the magnetic particle chamber 18 due to the movement of the combined body including the target substance by applying a magnetic force described later. The combined body can be suitably moved in the test liquid.

There are no restrictions on the fluorescent particles (phosphor, fluorescent substance), and various known ones that are used for detecting a target substance using fluorescence can be used. Examples include fluorescent dyes, quantum dots, rare earths, fluorescent pigments, and biological fluorescent molecules. Commercial products are also available for these. The fluorescent particles may be phosphorescent particles.
Depending on the type of the target substance, the target substance may generate fluorescence when irradiated with excitation light. In this case, the target substance also serves as the fluorescent particles.

There is no limitation on the method for binding the target substance to the magnetic particles, and further, the method for binding the target substance to the fluorescent particles, and a known method according to the type of the target substance and the magnetic particles and the target substance and the fluorescent particles. Is available. Examples include physical adsorption, antigen-antibody reaction, DNA hybridization, biotin-avidin bond, chelate bond, amino bond, and the like.

Physical adsorption is a method of binding a target substance and magnetic particles using electrostatic bonding force such as hydrogen bonding. Physical adsorption is easy to implement because it does not require treatment of magnetic particles or the like. On the other hand, in the physical adsorption, the magnetic particles and the fluorescent particles do not specifically adsorb to the target substance, so the selectivity is low. That is, in physical adsorption, there is a possibility that magnetic particles and / or fluorescent particles bind to substances other than the target substance contained in the analyte.
On the other hand, since the antigen-antibody reaction utilizes specific binding with a target substance, there is an advantage that magnetic particles and fluorescent particles can be selectively bound to the target substance.

When using the antigen-antibody reaction, if the target substance is an antigen such as a virus, it is necessary to bind the antibody against the target substance to the magnetic particles and / or fluorescent particles in advance. There is. That is, when using the antigen-antibody reaction, if the target substance is an antigen such as a virus, it is necessary to modify the magnetic particles and / or fluorescent particles with an antibody against the target substance virus in advance. There is.

When both the magnetic particles and the fluorescent particles are bound to the target substance, it is preferable that at least one of the bonds is specific binding with the target substance as in the antigen-antibody reaction.
When binding both magnetic particles and fluorescent particles to the target substance, if both bonds are non-specific, both the magnetic particles and fluorescent particles bind to foreign substances other than the target substance. In this case, there is an inconvenience that it becomes impossible to distinguish between the target substance and the foreign substance.

In the present invention, magnetic particles that emit fluorescence when irradiated with excitation light and magnetized fluorescent particles can also be used.
In this case, the binding between the magnetic particle that emits fluorescence when irradiated with excitation light, and the target substance and the particle is preferably a specific bond with the target substance, such as an antigen-antibody reaction. When the binding between the target particle and the magnetic particles that emit fluorescence when irradiated with excitation light is nonspecific, the magnetic particles that emit fluorescence when irradiated with excitation light are bound to foreign substances other than the target material. In this case, there is a disadvantage that it becomes impossible to distinguish between the target substance and the foreign substance. The same applies to the fluorescent particles having magnetism.

As described above, the sample processing container 10 in the illustrated example has the rectangular cylindrical container main body 12 that is divided into the liquid chamber 16, the magnetic particle chamber 18, and the immobilization chamber 20.
In addition, in the region corresponding to the liquid chamber 16 of the container main body 12, a square tube-shaped inner tube 24 whose both surfaces are open is inserted to form a double tube structure, and between the container main body 12 and the inner tube 24. Is a fluorescent particle supply chamber 28. Accordingly, the liquid chamber 16 is inside the inner tube 24.

The container body 12 is a light transmissive container made of glass, resin, or the like.
As described above, the container main body 12 is a rectangular cylindrical (square column) container in which one end surface (upper surface) is open and the other end surface (bottom surface) is closed.

In the subject processing container of the present invention, the container body is divided into a chamber for containing a liquid for dissolving and / or dispersing the subject, a chamber for containing magnetic particles, and a substance for immobilizing the liquid. As long as a chamber for storing the liquid, preferably a chamber for storing a liquid containing fluorescent particles, can be provided, the shape is not limited, and any of various shapes can be used.
Therefore, the container main body (subject processing container) can be used in various shapes such as a rectangular tube, a hexagonal tube, and the like, a cylindrical shape, an elliptical tube shape, a regular tetrahedron, and a spherical shape. is there. Further, the container body may have a convex shape such as a curved surface (spherical shape), such as a test tube, a convex surface on the bottom surface, a concave surface on the bottom surface, and a concave surface on the bottom surface. You may have.
Further, the container body 12 has protrusions, convex portions, ribs, concave portions, and the like on the outer surface for the purpose of positioning a loading position in a detecting device, which will be described later, fixing to the detecting device, and forming a gripping portion. You may have.

In the container body 12, only the magnetic particle chamber 18 in which detection of a target substance to be described later is performed may be light-transmitting, and the other chambers may be light-shielding.
Further, in the magnetic particle chamber 18, only the region corresponding to the optical path of excitation light and the optical path of measurement light (fluorescence measurement optical path) in detection of a target substance to be described later is light-transmitting, and the other areas are light-shielding. It may be.

Similarly to the container body 12, the inner tube 24 is formed of glass, resin, or the like, and is a rectangular tube having both ends open.
The inner tube 24 does not necessarily need to be light transmissive. Moreover, there is no restriction | limiting also in the shape of the inner tube | pipe 24, If it can be inserted in the container main body 12 and a double tube can be formed, various shapes can be utilized.

As described above, the inside of the container body 12 is divided into the liquid chamber 16, the magnetic particle chamber 18, and the immobilization chamber 20 in the height direction of the rectangular cylinder by the partition walls 14a and 14b. The liquid chamber 16 and the magnetic particle chamber 18 are separated by a partition wall 14 a, and the partition wall 14 a becomes the bottom surface of the liquid chamber 16. On the other hand, the magnetic particle chamber 18 and the immobilization chamber 20 are separated by the partition wall 14 b, and the partition wall 14 b becomes the bottom surface of the magnetic particle chamber 18. The bottom surface of the immobilization chamber 20 is a closed end of the container body 12.
Further, an inner tube 24 is inserted into a region corresponding to the liquid chamber 16 of the container main body 12, and the bottom surface between the container main body 12 and the inner tube 24 is closed by the partition wall 14c, so that the fluorescent particle supply chamber 28 is formed. It is formed.

As described above, the partition wall 14a and the partition wall 14b are films that can be broken by a sample collection tool such as a swab. The partition wall 14 c is a film that can be broken by the breaking jig 30.
The partition wall 14a, the partition wall 14b, and the partition wall 14c have various functions as long as they have a sufficient function as the bottom surface of the corresponding chamber, and do not affect the target substance and the substance accommodated in the corresponding chamber. It can be formed of any material. As an example, the partition walls may be formed of various resin films (polymer films).
In addition, the partition wall 14a and the partition wall 14c may be a single film as long as they can be ruptured independently depending on the material, shape, thickness, thickness distribution, fixing method, and the like.

The liquid chamber 16 contains a liquid L1 that can dissolve and / or disperse a subject such as saliva.
Examples of the liquid L1 include phosphate buffer, Tris buffer, acetate buffer, citrate buffer, tartrate buffer, water, aqueous liquid, alcohol liquid, and the like. It is preferable to use pure water, ion exchange water, or distilled water as the water.
Note that the same liquid as the liquid L1 can be used for the liquid L2 containing the fluorescent particles contained in the fluorescent particle supply chamber 28.
The liquid L1 and the liquid used for the liquid L2 may be the same or different.

The fluorescent particle supply chamber 28 contains a liquid L2 containing fluorescent particles. When the target substance is an antigen such as a virus, the fluorescent particles are preferably antibody-modified fluorescent particles.
In the liquid L2 containing fluorescent particles, the fluorescent particles may be dissolved, dispersed, or present in both dissolved and dispersed states.
The amount of the fluorescent particles accommodated in the fluorescent particle supply chamber 28 is not limited, and a necessary and sufficient amount may be appropriately set according to the type of the subject and the type of the target substance.

The breaking jig 30 is a rod-shaped member made of, for example, metal and resin.
The breaking jig 30 in the illustrated example has a substantially L-shaped shape, but is not limited thereto, and may be a linear bar-shaped member.
The breaking jig 30 is not limited to a rod-shaped member as long as it can break the partition wall 14c, and various shapes of members formed of various materials can be used.

In the sample processing container 10 shown in the drawing, the fluorescent particle supply chamber 28 is formed inside the container main body 12. However, the present invention is not limited to this, and the fluorescent particle supply is performed outside the container main body. A chamber may be formed.
For example, a tubular body may be provided in communication with the magnetic particle chamber 18, and the tubular body and the magnetic particle chamber 18 may be separated by a breakable partition wall, whereby the tubular body may be used as the fluorescent particle supply chamber.

In the container body 12 (the subject processing container 10), a space between the partition wall 14a and the partition wall 14b is a magnetic particle chamber 18.
As described above, magnetic particles are accommodated in the magnetic particle chamber 18. When the target substance is an antigen such as a virus, the magnetic particles are preferably antibody-modified magnetic particles.
The amount of magnetic particles accommodated in the magnetic particle chamber 18 is not limited, and a necessary and sufficient amount may be set as appropriate according to the type of subject and the type of target substance.
In the specimen processing container 10, the magnetic particle chamber 18 serves as a detection chamber (detection cell) for detecting the target substance when detecting the target substance described later.

In the container body 12, an immobilization chamber 20 is provided below the magnetic particle chamber 18. In the immobilization chamber 20, a substance capable of immobilizing the liquid L1 accommodated in the liquid chamber 16 is accommodated.
In the subject processing method of the present invention, there is no limitation on the method for immobilizing the liquid L1. As an example, a method of immobilizing the liquid L1 by embedding the liquid L1 in the matrix is illustrated. In other words, a method of immobilizing the liquid L1 by enclosing the liquid L1 in the matrix is exemplified.
Therefore, immobilization of the liquid L1 may be gelation using a gelling agent (crosslinking agent) or absorption using an absorbent material. The gelation using the gelling agent includes curing using a curing agent (crosslinking agent).

The gelling agent may be a compound or a mixture that can form a gel by contact with the liquid L1. As an example of the gelling agent, gelling agent A that forms a non-covalent bond by contacting with liquid L1 and gels, and gel that forms a covalent bond by contacting with liquid L1 and gels. The agent B is mentioned.
Examples of the gelling agent A include a mixture of sodium alginate and a calcium compound. When the liquid L1 and the mixture are mixed, calcium ions derived from the calcium compound are released into the liquid L1, gelation of sodium alginate proceeds via the calcium ions, and the liquid L1 is immobilized in the gel. As the gelling agent A, in addition to those described above, a gelling agent capable of forming a non-covalent bond such as a hydrogen bond, a π-π interaction, and an ionic bond can also be used. More specifically, examples thereof include polysaccharides such as gelatin and carrageenan, synthetic polymers such as polyvinyl alcohol, and low molecular hydrogelators.
Examples of the gelling agent B include a compound or a mixture that can be dissolved in the liquid L1 to form a covalent bond. More specifically, a mixture of an isocyanate compound and an alcohol compound capable of forming a urethane bond, a compound capable of proceeding an addition reaction such as a Michael addition reaction, and a compound capable of proceeding a condensation reaction.

Further, the absorbent material is an absorbent material that can swell and absorb the liquid L1, for example, a polymer having a three-dimensional cross-linked structure. More specifically, a polymer obtained by crosslinking polyvinyl alcohol, polyacrylic acid, or the like can be given.

In the specimen processing container 10 of the present invention, the immobilization chamber 20 may have a disinfectant and / or a disinfectant in addition to a substance capable of immobilizing the liquid L1 as necessary.
Since the immobilization chamber 20 has a bactericidal agent and / or a disinfectant, the contamination with the target substance when the target substance is bacteria, viruses, etc., and the surplus target substance flows into the immobilization chamber 20 is more preferable. Can be prevented.
The bactericidal agent and the disinfectant are not limited, and those that can inactivate, decompose, sterilize, etc. the target substance may be appropriately selected according to bacteria, viruses, and the like that are the target substance. Examples of bactericides include reverse soap bactericides such as benzalkonium chloride and cetylpyridium chloride, chlorine bactericides such as sodium hypochlorite, alcohol bactericides such as aqueous ethanol, and cresol. And phenolic fungicides such as

The subject processing method of the present invention will be described in detail below by describing the processing of the subject using the subject processing container 10 shown in FIG. 1 with reference to the conceptual diagrams of FIGS.
In the following description, the specimen processing container is also simply referred to as a processing container. Further, in order to simplify the drawings, the container body 12 and the inner tube 24 are indicated by bold lines in FIGS. 2 and 3.

In the processing of an object using the processing container 10, as an example, first, the processing container 10 is loaded into a target substance detection device.
As will be described later, as an example, the detection device detects the emission (fluorescence) by the fluorescent particles by irradiating the excitation light while moving the conjugate of the target substance, the magnetic particles, and the fluorescent particles by magnetic force. Detecting the target substance in the subject.

As described above, when detecting the target substance, the magnetic particle chamber 18 serves as a detection chamber for detecting the target substance. Therefore, as shown in FIGS. 2 and 3, when the processing container 10 is loaded at a predetermined position of the detection device, it faces the magnetic particle chamber 18 so as to face the magnetic particle chamber 18. The electromagnet 50 and the electromagnet 52 of the detection device are located.
The electromagnet 50 and the electromagnet 52 are known electromagnets having a coil, an iron core, a power source, a switch, and the like.

When the processing container 10 is loaded at a predetermined position of the detection apparatus, the sample is dissolved in the liquid L1 by immersing the swab S from which the sample is collected in the liquid L1 in the liquid chamber 16 and vibrating the swab S. Or disperse.

When the analyte is dissolved and / or dispersed in the liquid L1, the partition 14a is then broken by the swab S as shown in the second from the left in FIG. 2, and the liquid L1 containing the analyte is magnetized by dropping due to gravity. It flows into the particle chamber 18.
As described above, magnetic particles are accommodated in the magnetic particle chamber 18. When the liquid L1 containing the analyte flows into the magnetic particle chamber 18, the magnetic particles are bonded to the target substance, and a combined body of the target substance and the magnetic particles is formed.

When the liquid L1 in which the specimen is dissolved in the magnetic particle chamber 18 flows into the magnetic particle chamber 18, the electromagnet 50 and the electromagnet 52 of the detection device are driven. As a result, as shown in the third view from the left in FIG. 2, the combined body of the target substance and the magnetic particles and the excess magnetic particles are attracted to the electromagnet 50 and the electromagnet 52, and the magnetic particle chamber 18 ( Adsorbed to the inner wall surface of the container body 12). In FIG. 2 and FIG. 3, the adsorbent adsorbed on the inner wall surface of the magnetic particle chamber 18 by this magnetic force is indicated by the symbol C.

When a predetermined time has elapsed after driving the electromagnet 50 and the electromagnet 52, as shown on the right side of FIG. 2, the partition 14b is broken by the swab S, and the liquid L1 containing the subject is fixed by dropping due to gravity. It is made to flow into the chemical conversion chamber 20. When the partition wall 14 b is broken, the swab S is extracted from the processing container 10.
In the present invention, as described above, the conjugate of the target substance and the magnetic particles is separated from the liquid containing the analyte by using the magnetic force. Therefore, all the target substances existing in the magnetic particle chamber 18 serving as the detection chamber are combined with the magnetic particles.

As described above, the immobilization chamber 20 includes a substance that immobilizes the liquid L1 by gelation and absorption.
Therefore, as shown on the left side of FIG. 3, the liquid L1 that has flowed into the immobilization chamber 20 is immobilized by gelation or absorption, becomes an immobilization body SL, and does not flow.

As described above, according to the present invention, after a conjugate of a target substance such as a virus and a magnetic particle is generated in the liquid L1 containing the analyte, the fluid containing the conjugate and the analyte is obtained using magnetic force. And the liquid L1 containing the analyte from which the conjugate has been separated is immobilized. That is, the surplus target substance that is not bonded to the magnetic particles is separated from the bonded body and immobilized.
Therefore, as a preferred embodiment, the liquid L2 containing fluorescent particles is then supplied to the magnetic particle chamber 18 where the conjugate of the target substance and magnetic particles is present, so that the target substance, magnetic particles, and fluorescent particles Can be formed.
Therefore, according to the present invention, even when the target substance in the specimen is large, only the conjugate of the target substance and the magnetic particles and the conjugate of the target substance and the fluorescent particles are formed, and the target substance and the magnetic particles are formed. When the target substance is present in the specimen, it is possible to reliably detect the target substance. The method for detecting the target substance will be described in detail later.

Further, according to the present invention, a target substance and magnetic particles, preferably in addition to fluorescent particles, are combined in one processing container 10 (container body 12), and further, a target containing an excess target substance. The liquid L1 containing the specimen is immobilized.
Therefore, it is not necessary to transfer the liquid and the like, and the processing can be performed easily. Moreover, since a target substance such as a virus that may become a pollutant is immobilized in one container, the environment is not polluted by the target substance, and the processing container 10 is discarded after the inspection. However, the possibility of polluting the environment is very low. In particular, since the immobilization chamber 20 has a disinfectant, a disinfectant, and the like, the possibility of causing environmental pollution due to the target substance can be further reduced. In the present invention, after the detection of the target substance, if necessary, the sterilization process and the sterilization process may be performed by heating, ultraviolet irradiation, ozone treatment, etc. before discarding the processing container 10. Good.

As shown on the left side of FIG. 3, when the liquid L1 flowing into the immobilization chamber 20 is immobilized and becomes the immobilization body SL, the driving of the electromagnet 50 and the electromagnet 52 is stopped. Further, as shown in the center of FIG. 3, the partition 14 c is broken by the breaking jig 30, and the liquid L <b> 2 containing fluorescent particles flows into the magnetic particle chamber 18.
At this time, the partition wall 14b which was the bottom surface of the magnetic particle chamber 18 is broken, but the immobilization body SL in which the liquid L1 containing the subject is immobilized is present in the immobilization chamber 20, so this immobilization is performed. The body SL becomes the bottom surface of the magnetic particle chamber 18. That is, according to the processing container 10, it is not necessary to move the liquid or the like in order to mix the combined body of the target substance and the magnetic particles with the liquid L2 containing the fluorescent particles.

When the liquid L2 containing fluorescent particles flows into the magnetic particle chamber 18, the conjugate of the target substance and magnetic particles is mixed with the liquid L2 containing the fluorescent particles to form a conjugate of the target substance, magnetic particles and fluorescent particles. Is done. As described above, the target substances existing in the magnetic particle chamber 18 are all target substances bonded to the magnetic particles, and the surplus target substance is immobilized in the immobilization chamber 20. Therefore, according to the present invention, when the target substance is present in the subject, a combined body of the target substance, the magnetic particles, and the fluorescent particles can be surely formed.

In addition, in the magnetic particle chamber 18 in this state, in addition to the combined body of the target substance, the magnetic particles, and the fluorescent particles, the combined body of the target substance and the magnetic particles, the magnetic particles, and the liquid containing the fluorescent particles. Contained.

When the processing of the subject is completed in this way, the target substance is then detected in the magnetic particle chamber 18 by the detection device.
As described above, for example, the detection device detects the emission (fluorescence) by the fluorescent particles by irradiating the excitation light while moving the conjugate of the target substance, the magnetic particles, and the fluorescent particles by magnetic force. Detect the target substance.

Hereinafter, an example of a detection apparatus for detecting a target substance will be described with reference to the conceptual diagrams of FIGS. 4 and 5.
4 is a perspective view, and FIG. 5 is a view from above.

As shown in FIGS. 4 and 5, the detection device includes an excitation light irradiation unit 54 and an imaging unit 56 in addition to the electromagnet 50 and the electromagnet 52 described above.
As described above, the magnetic particle chamber 18 of the processing container 10 acts as a target substance detection chamber (detection cell) when the target substance is detected. Therefore, in a state where the processing container 10 is loaded in the detection device, the electromagnet 50 and the electromagnet 52 of the detection device correspond to the magnetic particle chamber 18 of the processing container 10 so as to face each other with the magnetic particle chamber 18 interposed therebetween. To position.

The excitation light irradiation part 54 is for irradiating the liquid accommodated in the magnetic particle chamber 18 with excitation light to generate fluorescence in the fluorescent particles.
In the illustrated detection apparatus, the excitation light irradiation unit 54 includes a light source 60 and a condensing optical system 62.

The light source 60 emits excitation light for generating fluorescence in the fluorescent particles. There is no restriction | limiting in the light source 60, Various light sources which can irradiate the light containing the component which excites a fluorescent particle and generate | occur | produces fluorescence can be utilized.
Examples of the light source 60 include a light bulb such as a mercury lamp, a fluorescent lamp, an LED (Light Emitting Diode), a laser such as a semiconductor laser, and the like. Among these, LEDs and semiconductor lasers are preferably used.

The condensing optical system 62 is for collecting the excitation light irradiated by the light source 60 and irradiating the inside of the magnetic particle chamber 18.
The condensing optical system 62 preferably has a focal point inside the magnetic particle chamber 18. The condensing optical system 62 preferably has a focal point at a position away from the inner wall of the magnetic particle chamber 18 by 100 nm or more, and more preferably has a focal point at a position away from the inner wall of the magnetic particle chamber 18 by 300 nm or more. .
The condensing optical system 62 is not limited, and various known condensing optical systems such as a condensing optical system using one or a plurality of lenses can be used.

The imaging unit 56 images the inside of the magnetic particle chamber 18.
In the illustrated detection apparatus, the imaging unit 56 includes an imaging element 64 and a condensing optical system 68.

The image sensor 64 is a known image sensor. Therefore, there is no restriction | limiting in the image pick-up element 64, According to the wavelength range of the fluorescence which a fluorescent particle generate | occur | produces, various image pick-up elements which have sensitivity to the light of this wavelength range can be utilized.
Examples of the imaging element 64 include a complementary metal oxide semiconductor (CMOS) image sensor (CMOS camera) and a charge-coupled device (CCD) image sensor (CCD camera). Among these, a CMOS image sensor and a CCD image sensor are preferably used.

The condensing optical system 68 is an optical system that condenses the optical path of photometry by the image sensor 64 inside the magnetic particle chamber 18 when viewed from the image sensor 64 side. That is, the condensing optical system 68 is a condensing optical system having a focal point inside the magnetic particle chamber 18.
The condensing optical system 68, that is, the imaging unit 56, preferably has a focal point inside the magnetic particle chamber 18. The condensing optical system 68 preferably has a focal point at a position away from the inner wall of the magnetic particle chamber 18 by 100 nm or more, and more preferably has a focal point at a position away from the inner wall of the magnetic particle chamber 18 by 300 nm or more. .
The condensing optical system 68 is not limited, and various known condensing optical systems such as a condensing optical system using one or a plurality of lenses can be used.

Note that the focal point of the excitation light irradiation unit 54 (condensing optical system 62) and the focal point of the imaging unit 56 (condensing optical system 68) may or may not coincide.

Here, in the detection device, the intersection of the optical axis (optical path) of the excitation light irradiation unit 54 and the optical axis (optical path of light to be measured) of the imaging unit 56 is located other than the inner wall surface of the magnetic particle chamber 18. Is preferable (see FIG. 5). That is, in the detection device, the intersection of the optical axis of the excitation light irradiation unit 54 and the optical axis of the imaging unit 56 is preferably located inside the magnetic particle chamber 18.
By adopting such a configuration for the detection apparatus, noise due to fluorescent particles adsorbed nonspecifically on the inner wall surface of the magnetic particle chamber 18 is removed, and a highly sensitive subject having a high S / N ratio. (Target substance) becomes possible. This will be described in detail later.

The intersection of the optical axis of the excitation light irradiating unit 54 and the optical axis of the imaging unit 56 is preferably located within the magnetic particle chamber 18 at a location away from the inner wall surface of the magnetic particle chamber 18 by 100 nm or more, It is more preferable to be located 200 nm or more away from the inner wall surface of the magnetic particle chamber 18, and more preferable to be located 300 nm or more away from the inner wall surface of the magnetic particle chamber 18.

The optical axis of the excitation light irradiation unit 54 is the optical axis of the light source 60 when the condensing optical system 62 is not provided, and when the condensing optical system 62 is provided, the optical axis of the condensing optical system 62 is used. The optical axis. In addition, the optical axis of the light source 60 and the optical axis of the condensing optical system 62 are normally made to correspond.
The optical axis of the imaging unit 56 is the optical axis of the imaging element 64 when the condensing optical system 62 is not provided, and the light of the condensing optical system 62 when the condensing optical system 62 is provided. Is the axis. Note that the optical axis of the image pickup element 64 and the optical axis of the condensing optical system 62 are usually matched.

Hereinafter, with reference to FIG. 6, the operation of detecting the target substance by the detection apparatus loaded with the processing container 10 will be described.
FIG. 6 is a view of the processing container 10 as viewed from the same direction as FIGS. In FIG. 6, the excitation light irradiation unit 54 and the imaging unit 56 are illustrated in order to simplify the drawing and make it easy to understand the movement of the combined body of the target substance, magnetic particles, and fluorescent particles (the detected combined body T). Is omitted, and only the combined body of the target substance, magnetic particles, and fluorescent particles is shown inside the magnetic particle chamber 18.

As described above, in the state in which the processing of the subject is completed, the magnetic particle chamber 18 of the processing container 10 has a conjugate of the target substance, magnetic particles, and fluorescent particles, a conjugate of the magnetic particles and the target substance, A liquid containing magnetic particles and fluorescent particles is contained.
In the following description, a combination of a target substance, magnetic particles, and fluorescent particles is also referred to as “detected combination T” for convenience.

In a state where the electromagnet 50 and the electromagnet 52 are not driven, the detected coupled body T, the coupled body of magnetic particles and the target substance, magnetic particles, fluorescent particles, and the like are in the magnetic particle chamber 18 (liquid). It is in an irregularly floating state.
When only the electromagnet 50 is driven from this state, the detected coupled body T, the coupled body of the magnetic particles and the target substance, and the magnetic particles move toward the electromagnet 50 by magnetic force. The fluorescent particles move due to Brownian motion or the like, but do not move due to magnetic force because they do not have magnetic particles.
Next, when driving of the electromagnet 50 is stopped and the electromagnet 52 is driven, the detected coupled body T, the coupled body of the magnetic particles and the target substance, and the magnetic particles move toward the electromagnet 52 by magnetic force.
Next, when driving of the electromagnet 52 is stopped and the electromagnet 50 is driven, the detected coupled body T, the coupled body of the magnetic particles and the target substance, and the magnetic particles move toward the electromagnet 50 by magnetic force.

That is, by alternately driving the electromagnet 50 and the electromagnet 52, the detected coupled body T, the coupled body of the magnetic particles and the target substance, and the magnetic particles in the magnetic particle chamber 18 are electromagnet 50 by the magnetic force. And the electromagnet 52.

As described above, when the excitation light irradiation unit 54 irradiates the inside of the magnetic particle chamber 18 with the excitation light while the electromagnet 50 and the electromagnet 52 are alternately driven, the fluorescent particles emit light. That is, when excitation light is irradiated into the inside of the magnetic particle chamber 18 by the excitation light irradiation unit 54, the detected coupled body T and the fluorescent particles emit fluorescence.
On the other hand, the combination of the magnetic particles and the target substance, and the magnetic particles do not change when irradiated with excitation light.

As described above, by alternately driving the electromagnet 50 and the electromagnet 52, the detected coupled body T reciprocates. However, since the fluorescent particles do not have magnetic particles, they are not moved by magnetic force.
On the other hand, the to-be-detected conjugate T and the fluorescent particles emit fluorescence by irradiation with excitation light, but the conjugate of the magnetic particles and the target substance that reciprocate similarly to the to-be-detected conjugate T, and the magnetic particles are: Since it does not have fluorescent particles, it does not emit light.
That is, only the detected coupled body T reciprocates while emitting fluorescence in a state where the electromagnet 50 and the electromagnet 52 are driven alternately and irradiated with excitation light.

Therefore, while the electromagnet 50 and the electromagnet 52 are driven alternately, the inside of the magnetic particle chamber 18 is irradiated with the excitation light and the inside of the magnetic particle chamber 18 is imaged by the imaging unit 56, so that the image is captured ( In the moving image), if there is light (bright spot) that reciprocates according to the alternating drive of the electromagnet 50 and the electromagnet 52, the detected coupled body T exists in the liquid in the magnetic particle chamber 18. That is, it can be detected that the target substance is present in the subject.
Here, even if the detected conjugate T (target substance) contained in the liquid in the magnetic particle chamber 18 is very small, the electromagnet 50 and the electromagnet 52 are alternately arranged as long as at least one detected conjugate T exists. The light reciprocates according to the drive. Therefore, even if the amount of the target substance in the subject is very small, the target substance can be detected, and the presence or absence of the target substance in the subject can be detected. That is, according to the specimen processing method of the present invention, if a target substance is present in the specimen, it is possible to reliably generate a detected conjugate T (a conjugate of the target substance, magnetic particles, and fluorescent particles). Therefore, if the target substance exists in the subject, the target substance can be reliably detected.

Such detection of the target substance may be performed in the vicinity of the wall surface of the magnetic particle chamber 18. That is, as described in Patent Document 1, a near field may be formed in the magnetic particle chamber 18 to detect the target substance. In other words, the intersection of the optical axis of the excitation light irradiation unit 54 and the optical axis of the imaging unit 56 may be located in the inner wall surface of the magnetic particle chamber 18 or in the vicinity of the inner wall surface.
However, there is a possibility that many foreign substances such as proteins are attached to the inner wall surface (near field) of the magnetic particle chamber 18, and further, fluorescent particles and magnetic particles are adsorbed nonspecifically. The smooth movement of the specimen is hindered, and the detection sensitivity of the detected conjugate T decreases.
Therefore, preferably, as described above, the target substance is detected inside the magnetic particle chamber 18. In other words, the intersection of the optical axis of the excitation light irradiation unit 54 and the optical axis of the imaging unit 56 is positioned other than the inner wall surface of the magnetic particle chamber 18, that is, inside the magnetic particle chamber 18. This avoids inconveniences that occur when the target substance is detected in the vicinity of the wall surface of the magnetic particle chamber 18, and enables detection of a highly sensitive analyte with a high S / N ratio.

In the target substance detection method described above, the detected conjugate T is detected by imaging using an image sensor such as a CCD image sensor, but the detection method of the detected conjugate T is not limited thereto.
For example, instead of the image pickup device, detection using a light amount measuring means (photosensor) such as a photodiode, a phototransistor, and a photomultiplier tube (photomultiplier) can be used.
When the light intensity measuring means is used and the detected coupled body T exists in the liquid as described above, when the detected coupled body T is reciprocated by the alternating drive of the electromagnet 50 and the electromagnet 52, the detected coupled body. In a state where T is outside the detection region by the light amount measuring means, the amount of light detected is small. When the detected coupled body T enters the detection region by the light amount measuring means by movement, the detected light amount increases.
Accordingly, all the detected coupled bodies T are once moved by magnetic force to the wall surface on the electromagnet 50 or electromagnet 52 side, and then the electromagnet 50 and the electromagnet 52 are alternately driven, thereby detecting the coupled bodies T to be detected. The amount of detected light increases or decreases according to the reciprocating motion of. That is, it is possible to detect whether or not the target substance is present in the liquid, that is, in the subject, by detecting the increase / decrease in the detected light amount.

The processing container 10 shown in FIG. 1 and FIG. 2 has a fluorescent particle supply chamber 28 for supplying the liquid L2 containing fluorescent particles, with the processing container 10 having a double tube structure as a preferred embodiment. However, the present invention is not limited to this.
For example, the processing container does not have the fluorescent particle supply chamber for supplying the liquid L2 containing the fluorescent particles, and after immobilizing the liquid L1 containing the subject, the target substance and the magnetic particles using a dropper or the like The liquid L2 containing the fluorescent particles may be supplied to the magnetic particle chamber 18 in which the combined body exists.

Further, the processing container 10 shown in FIG. 1 has the partition wall 14b, so that the magnetic particle chamber 18 and the immobilization chamber 20 are completely separated, and the partition wall 14b is broken by the swab S, so that the magnetic particle chamber Although 18 and the immobilization chamber 20 are communicated, the present invention is not limited to this.
For example, instead of the partition wall 14b that can be broken by the swab S, the magnetic particle chamber 18 and the immobilization chamber 20 are separated by a mesh, a nonwoven fabric, a filter, or the like, and the breaking by the swab S or the like is not performed. The liquid L1 containing the subject may fall from the magnetic particle chamber 18 to the immobilization chamber 20 due to gravity.

In the processing container 10 shown in FIG. 1, the magnetic particle chamber 18 has a chamber in which the target substance and the combination of magnetic particles and the liquid L1 containing the target object are separated by a magnetic force, and detection of the target substance by the detection device. It becomes a detection chamber when performing. However, the present invention is not limited to this, and includes a magnetic particle chamber 18 in which magnetic particles are mixed into the liquid L1 containing the subject, and a chamber in which a target substance is detected by a magnetic separation and detection device. It may be divided.
For example, like the (subject) processing container 70 shown in FIG. 7, a separation detection chamber 72 separated by the partition 14 d is provided below the magnetic particle chamber 18, and the partition 14 b described above is provided below the separation detection chamber 72. A configuration in which the immobilization chamber 20 separated in (1) is provided can also be used.
The partition 14d may be a mesh, a nonwoven fabric, a filter, or the like, similar to the partition 14b described above.

In the processing container 70, as in the processing container 10 described above, after the sample collected by a swab or the like in the solution chamber 14 is dissolved and / or dispersed in the liquid L 1, the liquid L 1 is caused to flow into the magnetic particle chamber 18. Forming a conjugate of the target substance and magnetic particles.
Next, the partition wall 14d is broken by a swab, and the liquid L1 containing the conjugate of the target substance and the magnetic particles and the analyte is caused to flow into the separation detection chamber 72.
When the liquid L1 flows into the separation detection chamber 72, the electromagnet 50 and the electromagnet 52 are driven to adsorb the combined substance of the target substance and magnetic particles to the inner wall of the separation detection chamber 72 in the same manner as the processing container 10 described above. Let Next, the partition wall 14b is broken by a swab or the like, and the liquid L1 containing the specimen is caused to flow into the immobilization chamber 20 to be immobilized.
Next, the partition 14c is broken by the breaking jig 30, and the liquid L2 containing fluorescent particles is caused to flow into the separation detection chamber 72 through the magnetic particle chamber 18 to form a combined body of the target substance, magnetic particles, and fluorescent particles. To do. Thereafter, as described above, the target substance is detected by the detection device.

In the processing container 70 shown in FIG. 7, the partition wall 14d and the partition wall 14b are broken by a swab to cause each liquid to flow into the next chamber (lower chamber). However, the present invention is not limited to this.
As an example, a (subject) container 76 shown in FIG. 8 is illustrated. The storage container 76 has a partition wall 14e through which a liquid L1 containing a specimen such as a mesh, a nonwoven fabric, and a filter can be passed, instead of the partition wall 14d that can be broken by a swab. Further, a packing 78 made of rubber or the like is provided on the lower surface (bottom wall) of the container main body 12, and the breaking jig 80 is provided by the packing 78 by being held fluid-tight by a breaking jig.

In the processing container 70, first, similarly to the processing container 70 described above, the liquid L <b> 1 containing the subject flows into the magnetic particle chamber 18.
When the liquid L1 containing the subject is in the magnetic particle chamber 18, a combined body of the target substance and the magnetic particles is formed. In parallel, the liquid L1 containing the conjugate of the target substance and magnetic particles and the analyte passes through the partition wall 14e and flows into the separation detection chamber 72 by gravity.
When the liquid L1 flows into the separation detection chamber 72, the electromagnet 50 and the like are driven in the same manner as the processing container 70 described above, and the combined body of the target substance and the magnetic particles is adsorbed on the inner wall of the separation detection chamber 72.
Next, the partition 14b is broken from below by the breaking jig 80, and the liquid L1 containing the specimen flows into the immobilization chamber 20 and is immobilized.
Thereafter, similarly, the liquid L2 containing fluorescent particles is caused to flow into the separation detection chamber 72 to form a combined body of the target substance, magnetic particles, and fluorescent particles, and the target substance is detected by the detection device.

In the above example, a plurality of chambers are arranged in the vertical direction (vertical direction) and the liquid is moved by gravity drop. However, the present invention is not limited to this, and a plurality of chambers are arranged in the horizontal direction (horizontal direction). A configuration in which the chambers are arranged can also be used.
In this case, an immobilized body in which the liquid L1 containing the subject is immobilized may be used as, for example, a side wall that forms the detection chamber.

In the present invention, it is preferable to use a blocking agent in combination.
By using a blocking agent, it is possible to prevent aggregation of magnetic particles, fluorescent particles, and conjugates of target substances, magnetic particles and fluorescent particles, and more appropriate treatment of analytes and target substances after treatment Can be detected.
In addition, as a blocking agent, well-known various things can be utilized. As an example, bovine serum albumin (BSA (Bovine Serum Albumin)), casein, skim milk, polyethylene glycol and the like are exemplified.

In the present invention, the supply position of the blocking agent to various liquids is not limited, and various places in the processing container can be used.
For example, in the processing container 10 shown in FIG. 1, the liquid chamber 16 may have a blocking agent, the magnetic particle chamber 18 may have a blocking agent, and the fluorescent particle supply chamber 28 has a blocking agent. Any two of these chambers may have a blocking agent, and all three chambers may have a blocking agent.

As described above, the sample processing method and the sample processing container of the present invention have been described in detail. However, the present invention is not limited to the above-described examples, and various improvements and modifications can be made without departing from the gist of the present invention. Of course, you may do this.

It can be suitably used for detection of viruses and the like in medicine and research.

10, 70, 76 (Subject) Processing container 12, 78 Container body 14a, 14b, 14c, 14d, 14e Partition 16 Liquid chamber 18 Magnetic particle chamber 20 Immobilization chamber 24 Inner tube 28 Fluorescent particle supply chamber 30, 80 Break Jig 50, 52 Electromagnet 54 Excitation light irradiation unit 56 Imaging unit 60 Light source 62, 68 Condensing optical system 64 Imaging element 72 Separation detection chamber 78 Packing L1, L2 Liquid SL immobilized body T Detected assembly

Claims (17)

Supplying a specimen into a liquid in a container to form a conjugate in which the target substance and magnetic particles in the specimen are bound in the liquid;
In the container, the conjugate and the liquid containing the analyte are separated by magnetic force,
A method for processing a specimen, comprising: separating the conjugate and the liquid containing the specimen, and then immobilizing the liquid containing the specimen in the container. The method for treating an analyte according to claim 1, wherein after the liquid containing the analyte is immobilized, fluorescent particles are bound to the conjugate. The method for processing an analyte according to claim 2, wherein the binding of the conjugate and the fluorescent particles is performed in a space in which a fixed body on which a liquid containing the analyte is immobilized is a part of a wall surface. 4. The object processing method according to claim 3, wherein the immobilized body is a bottom surface of the space. 5. The method according to claim 2, wherein the binding of the conjugate and the fluorescent particles is performed by supplying the liquid containing the fluorescent particles to the conjugate separated from the liquid containing the analyte. The processing method of the test object of description. The liquid containing the fluorescent particles is stored in a supply chamber in which the wall surface of the container is a part of its wall surface,
The method for processing a subject according to claim 5, wherein the supply of the liquid containing the fluorescent particles to the conjugate separated from the liquid containing the subject is performed by breaking a part of the supply chamber. The object processing method according to claim 6, wherein the container is tubular, and the supply chamber has a double-tube structure in which the wall of the container is a single tube. The subject processing method according to claim 6 or 7, wherein the inner wall surface of the container is a part of the wall surface of the supply chamber. The method for processing a subject according to any one of claims 1 to 8, wherein the container is separated into a plurality of chambers by walls that can be broken by the sample collection tool. The object processing method according to claim 9, wherein the substance is moved between the plurality of chambers by gravity. One of the plurality of chambers is a liquid chamber containing the liquid,
Another one of the plurality of chambers is a magnetic particle chamber containing the magnetic particles,
The subject processing method according to claim 9 or 10, wherein another one of the plurality of chambers is an immobilization chamber for immobilizing the liquid containing the subject. The said container WHEREIN: The said liquid chamber, the said magnetic particle chamber, and the said fixation chamber are provided in order of the said liquid chamber, the said magnetic particle chamber, and the said fixation chamber toward one direction. Analyte processing method. The method for processing an object according to claim 11 or 12, wherein the liquid chamber and the magnetic particle chamber, and the magnetic particle chamber and the immobilization chamber are adjacent to each other. The specimen processing method according to any one of claims 11 to 13, wherein the immobilization chamber has at least one of a disinfectant and a disinfectant. The method for processing an object according to any one of claims 1 to 14, wherein the container has a blocking agent. A container for processing a subject,
The interior is separated into a plurality of chambers by walls that can be broken by the subject sampling tool, and one of the plurality of chambers contains a liquid supplied to the subject, One of the chambers contains magnetic particles, and the other one of the plurality of chambers contains a substance for immobilizing the liquid. . The specimen processing container according to claim 16, further comprising a chamber for storing fluorescent particles in at least one of the inside of the container and the outside of the container.

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