JP3865134B2 - Biochip cartridge - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a biochip cartridge for testing biopolymers such as DNA, RNA (mRNA, cDNA, etc.) and protein, and more particularly to a biochip cartridge that is highly safe and can reduce test costs. is there.
[0002]
[Prior art]
A method for testing DNA or the like using a biochip has been well known. FIG. 8 shows an example of a conventional cartridge for biochip (also simply referred to as biochip) that scans a hybridized DNA chip with a biochip reader and reads an unknown DNA sequence (see, for example, Patent Document 1). ).
[0003]
In this apparatus, the biochip reader 20 irradiates the hybridized DNA chip in the biochip 10 with excitation light and reads the fluorescence generated from the fluorescent label to detect an unknown DNA sequence and the like. The container 11 is made of a material transparent to the excitation light and fluorescence.
[0004]
The biochip 10 in this case is one in which a substrate 12 having a large number of known DNA chip sites CL arranged in an array is stored in a container 11 as shown in FIG. Prior to the reading operation, the biochip 10 is injected with a solution 15 containing an unknown DNA fragment previously attached with a fluorescent label from an introduction port 13 using a solution introduction means 14 such as a syringe as shown in FIG. Hybridize with a DNA chip.
[0005]
However, blood, which is a sample to be examined, may be contaminated with viruses such as HIV, and for safety, medical instruments such as syringes are used as disposable instruments. In the solution introduction method as shown in FIG. 10, since the solution transfer operation from the solution introduction means 14 to the container 11 is performed, there is a risk of contact with the solution due to an erroneous operation and infection with HIV or the like.
In addition, there are problems that the medical instruments to be discarded due to the disposable increase in the number of syringes, instruments used for pretreatment, solution introduction means, DNA chips, and the like, resulting in high inspection costs.
[0006]
FIG. 11 solves this point. The blood collection tube 31 is inserted into a syringe instead of a conventional Spitz tube to collect blood, and is formed in a cylindrical shape with a solid material transparent to excitation light and fluorescence. The opening of the blood collection tube 31 is sealed with a rubber stopper 32 in which a needle is punctured at the center, and the whole blood collection tube 31 is under negative pressure.
[0007]
The blood collected through the needle is once held in the collection unit 33 and then guided to the pretreatment unit 34 for pretreatment. The pretreatment is a series of processes such as separating lymphocytes from blood, extracting DNA from the separated lymphocytes, and adding a fluorescent label to the extracted DNA.
[0008]
A substrate 35 on which known DNA is arranged in an array is housed at the innermost part of the blood collection tube 31, and the DNA permeating from the pretreatment unit 34 and the known DNA are stored. Hybridization with the DNA of is carried out.
[0009]
However, although such a biochip has an advantage that blood collection, pretreatment, hybridization, etc. are performed automatically and consistently, a solid blood collection tube is necessary and expensive, and air for making negative pressure There is a problem in that a suction pump or the like is required and the whole is expensive.
[0010]
Japanese Patent Application Laid-Open No. 2002-365299 filed by the present applicant describes a biochip that solves this problem. This biochip has a structure as shown in FIG. This biochip 40 is formed into a flat sealed bag shape with a material that is flexible and transparent to excitation light and fluorescence.
[0011]
As shown in the plan view of FIG. 12 (b), the blood collection bag 41 has a square outer shape, its peripheral part is joined in a sealed manner, and its central part is a fish-shaped bag. The opening of the bag corresponding to the fish mouth is sealed with a stopper 42. The stopper 42 is formed of a rubber-like material, and an injection needle is inserted through the stopper when blood is collected. When the injection needle is removed after blood collection, the needle hole is immediately closed, and the collected blood does not leak to the outside.
[0012]
In the blood collection bag 41, a collection part 43, a pretreatment part 44, a coupling part 45, and a waste liquid storage part 47 are formed in this order from the stopper 42 to the back.
The collected blood is stored in the collection unit 43. Hooks 431 are formed on the front and back surfaces of the membrane of the collection part 43, respectively, and the collection part 43 is inflated by pulling the engaging member engaged with the hook 431 outward during blood collection.
[0013]
The preprocessing unit 44 performs a process of extracting an unknown target sample from the collected blood. The coupling unit 45 includes a substrate 46 on which a plurality of known samples (here, DNAs) are arranged in an array, and the sample extracted by the pretreatment unit 44 can be complementarily coupled to the known sample. It is like that.
The waste liquid storage unit 47 is a bag portion provided for storing unnecessary solutions pushed out from the pretreatment unit 44 and the coupling unit 45, and the bag is compressed in an initial state.
[0014]
On both sides of the pre-processing portion 44, bag portions 48 and 50 corresponding to the dorsal fin and the belly fin are formed in a relationship opposite to each other. In the bag portions 48 and 50, solutions necessary for extracting an unknown sample (DNA, RNA, protein, or the like) from blood are sealed, respectively.
Valves 49 and 51 for partition walls are formed at connecting portions (narrow passages) between the bag portions 48 and 50 and the pretreatment portion 44, and are formed so as to be broken when the pressure of the solution in the bag portion increases.
[0015]
After the collected blood is stored in the collection part 43 of the voting bag 41, the blood collection bag 41 is sandwiched between rotating rollers 61 and 62 as shown in FIG. go.
The rollers 61 and 62 have a length in the axial direction longer than the width of the blood collection bag 41, and press the entire width of the blood collection bag 41 uniformly.
[0016]
The collected blood is pushed to the pretreatment unit 44 by the rotation of the rollers 61 and 62. When the positions of the rollers 61 and 62 advance and the bag portion 48 starts to be crushed, the pressure in the bag portion 48 increases and the valve 49 is broken. When the valve 49 is broken, the solution in the bag portion 48 flows into the pretreatment portion 44, and a predetermined treatment with the solution is performed.
Subsequently, when the bag portion 50 is also crushed by the rollers 61 and 62, the valve 51 is similarly broken, and the solution in the bag portion 50 flows into the pretreatment portion 44, and a predetermined process is performed.
[0017]
Therefore, the time difference process can be easily performed by shifting the attachment position of the bag portion. That is, a process of separating lymphocytes from blood and extracting DNA from the separated lymphocytes, a process of adding a fluorescent label to the extracted DNA, and the like can be performed while being shifted in time.
[0018]
When the processing in the preprocessing unit 44 is completed, the rollers 61 and 62 are subsequently rotated. As a result, the processed blood is sent to the coupling portion 45 and hybridized with a known DNA chip placed on the substrate 46.
Excess blood and solution pushed out from the pretreatment unit 44 accumulate in the waste liquid storage unit 47. The hybridized DNA chip is read out by a biochip reading device (not shown) as in the prior art.
[0019]
In this manner, the processes from blood collection to pretreatment and hybridization are performed consistently in a sealed blood collection bag, and accidents such as accidental contact with the solution can be prevented. In addition, since such a blood collection bag is easily made of a soft, flexible and inexpensive material, an inexpensive biochip can be easily realized.
[0020]
[Patent Document 1]
JP 2001-235468 A (2nd page-5th page, FIG. 1, FIG. 4-6)
[0021]
[Problems to be solved by the invention]
However, such conventional biochips have the following problems.
(1) Although the bag 41 is crushed by the rollers 61 and 62, the bag is not uniformly crushed and the flow of blood and solution is difficult to stabilize.
(2) The bag 41 is soft and easily emits autofluorescence due to an adhesive, coating, plasticizer, and the like. This self-fluorescence becomes background noise and the S / N changes, so that it is impossible to detect weak fluorescence (weak signal) from a fluorescent substance labeled on DNA.
[0022]
An object of the present invention is to solve the above-described problems, and to realize a biochip cartridge that stabilizes blood and solution feeding and is less prone to self-fluorescence by using a hard material on the surface. is there.
[0023]
[Means for Solving the Problems]
In order to achieve such an object, the invention of claim 1
A flat substrate member made of an elastic material;
A flexible cover having a material harder than the substrate member and attached in close contact with the upper and lower surfaces of the substrate member,
The substrate member includes a collection unit for storing biopolymers, a pretreatment unit for pretreating the biopolymers, and a biological height prepared in advance from the pretreated biopolymers. A detection unit for detecting a biopolymer that binds to a molecule, and each gap portion of a flow path connecting the collection unit, the pretreatment unit, and the detection unit,
The biopolymer may be sequentially moved from the collection unit side through the pretreatment unit to the detection unit side.
[0024]
As a result, the processes from the injection of the biological sample to the hybridization are performed consistently in the sealed cartridge, and accidents such as contact with the injected solution due to erroneous operation can be prevented.
Further, it can be manufactured with an inexpensive material and is easy to manufacture, and an inexpensive biochip cartridge can be easily provided.
Furthermore, since the flow path is fixed, the remaining sample and uneven pressing are reduced, and accurate feeding can be performed. In addition, since a hard cover is used for pressing the substrate member, there is an advantage that autofluorescence from the cover hardly occurs.
[0025]
In this case, the movement of the sample containing the biopolymer, as described in claim 2, presses the cover with a roller-like rigid body, and crushes the voids of the substrate member from the collection unit to the detection unit side. Done by going.
[0026]
Further, in order to perform pretreatment on the sample, a pretreatment liquid stored in the bag portion by providing a bag portion filled with the pretreatment liquid on the substrate member and pressing the roller with a roller as in claim 3 Is sent to the pre-processing unit.
[0027]
Further, the substrate member is provided with a waste liquid storage part for storing the waste liquid discharged from the detection part.
Further, the cover may be structured to be attached to the front surface or the front surface and the back surface of the substrate member as in the fifth aspect.
[0028]
Further, the flow path formed in the substrate member is formed such that the gap is crushed by pressing the cover with a roller-like rigid body, as in claim 6.
Further, the cover can be made of plastic, glass or quartz.
Furthermore, when the biopolymer is optically detected, a cover formed of a transparent material is used so that the cover can be optically detected at least in the detection portion.
[0029]
Further, as in claim 9, a plurality of bag portions for storing the pretreatment liquid are arranged at different positions. Thus, when the substrate member is crushed by the roller-like rigid body, the pretreatment liquid flows from each bag portion into the pretreatment portion with a time difference.
Further, as in claim 10, the substrate member can be formed in a wedge shape in which the thickness gradually decreases from the sampling portion side toward the detection portion side.
[0030]
Further, as in claim 11, a valve for stopping the flow of the solution is provided in the flow path, and the valve can be formed to open when pressure is applied to the solution flowing in the flow path.
The substrate member can also be formed of a plastically deformable material or gel as in the twelfth aspect.
[0031]
DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, the present invention will be described in detail with reference to the drawings. FIG. 1 is a block diagram showing an embodiment of a biochip cartridge according to the present invention. 1A is a side view, FIG. 1B is a plan view of the substrate member, and FIG. 1C is a cross-sectional view taken along the line AA ′ in FIG. 1B (including a cover).
[0032]
In the biochip cartridge 100 of FIG. 1 (a), 101 and 102 are transparent and flexible covers such as hard plastic, and 110 is a substrate formed of an airtight and elastic material such as elastic rubber. It is a member. Covers 101 and 102 are attached to the upper and lower surfaces of the substrate member 100 in a sealed manner, for example, by bonding.
[0033]
As shown in FIG. 1 (b), the substrate member 110 is formed with a through-hole for the inlet channel 111, and has a plurality of chambers, that is, the sampling part 112, the first and second bag parts 113. 114, the pretreatment unit 115, the detection unit 116, the waste liquid storage unit 117, and the respective through holes for the flow path 118 that connect them to each other.
By bonding the covers 101 and 102 to the front and back of the substrate member 110, each hole is closed from above and below. For example, the flow path 118 is a void having a shape as shown in FIG.
[0034]
Each flow path and room will be further described in detail. The passage 111 is a sample injection path for injecting a solution containing a collected biopolymer such as blood (hereinafter simply referred to as a sample). It should be noted that by utilizing the fact that the substrate member 110 itself is a rubber-like elastic body, the passage 111 is not provided, and the sample may be directly injected into the collection member 112 by inserting the injection needle into the substrate member 110. I do not care.
[0035]
The collection unit 112 is a room for collecting collected samples such as collected blood. The first and second bag portions 113 and 114 store pretreatment liquids for separating, purifying, and amplifying the biopolymer to be detected from the sample of the collection unit 112.
[0036]
In the pretreatment unit 115, the sample and the pretreatment liquid from the bag portions 113 and 114 are mixed to perform pretreatment such as separation, purification, and amplification. The detection unit 116 includes an array chip (not shown) to which a biopolymer is fixed, and the biopolymer of the pretreated sample is complementarily bonded (hybridized) to the biopolymer, thereby It is a room to detect.
The waste liquid storage unit 117 is a room for storing waste liquid discharged from the detection unit 116 after hybridization.
[0037]
Next, a method of using and operating the biochip cartridge 100 formed as described above will be described. After injecting the sample into the collecting unit 112, the biochip cartridge 100 is placed on, for example, a flat plate (not shown), and a cylindrical roller-like rigid body (from the top of the cover 101 (see FIG. 2A)). (Hereinafter simply referred to as a roller) 200 is pressed in parallel with the cover, and is rotated from the inlet side toward the pretreatment unit 115.
[0038]
When the roller 200 is pressed, the cover 101 is bent and the substrate member 110 is crushed. As a result, the flow path 111 immediately below the center of the roller 200 is narrowed and closed, and a temporary valve is formed. When the roller 200 is rotated and moved to the right of the paper surface, the valve also moves to the right. This valve has the effect of preventing backflow.
[0039]
When the collecting unit 112 is crushed by the roller 200, the sample stored in the collecting unit 112 is pushed rightward and sent to the preprocessing unit 115 through the flow path 118.
[0040]
Next, when the bag portion 113 is similarly crushed by the roller 200, the pretreatment liquid is sent to the pretreatment portion 115 through the flow path 118. As the roller 200 further advances to the right, the bag portion 114 is similarly crushed, and the pretreatment liquid accumulated therein is also fed into the pretreatment portion 115. As a result, the pretreatment liquid is mixed into the sample in the pretreatment unit 115, and pretreatment such as separation, purification, and amplification of the biopolymer is performed.
As shown in FIG. 1B, the time difference process can be easily performed by shifting the positions of the bag portions 113 and 114 to the right in the drawing.
[0041]
When the preprocessing is completed, the roller 200 is rotated, the preprocessing unit 115 is crushed, and the processed sample is sent to the detection unit 116. In the detection unit 115, hybridization between the biopolymer in the sample and the biopolymer fixed to the array chip is performed. The biopolymer and the solution that have not been hybridized are sent to the waste liquid storage unit 117 by rotating the roller 200 to the right or tilting the entire biochip cartridge.
[0042]
The array chip subjected to hybridization can be read through the transparent cover 101 using a known biochip reading device (not shown).
[0043]
The present invention is not limited to the above-described embodiments, and includes many changes and modifications without departing from the essence thereof.
For example, as shown in FIG. 3, a shutter 210 is provided in addition to the roller 200, and by pressing this shutter 210, the flow path can be appropriately blocked to prevent the sample from proceeding inadvertently. .
[0044]
Further, the cartridge 100 may be sandwiched from above and below by two rollers, and the sample and the pretreatment liquid may be sent out as in the above embodiment.
In addition, as shown in FIG. 4, a stopper 119 is provided in the flow path 118 at the outlet of the sampling unit 112, and when the sample is injected into the sampling unit 112, it is blocked, and then the sampling unit 112 is crushed by a roller. For the first time, the stopper 119 is broken to open the flow path, and the sample may flow out of the flow path 118.
[0045]
Further, the thickness of the substrate member 110 is not uniform, and as shown in FIG. 5, as shown in FIG. 5, the substrate member 110 may have a wedge shape in which the collection portion side is thick and the waste liquid storage portion side is thin. As a result, the shape of the flow path and the bag portion can be changed depending on the location, so that the degree of freedom in design increases.
Further, each room or flow path of the substrate member 110 may be a concave hole as shown in FIG. 6 instead of the through hole. Further, the structure shown in FIG. 7 may be used in which the lower cover 102 is removed.
[0046]
The upper and lower covers may be glass plates or quartz plates. In addition, when the hybridized biopolymer can be electrically detected, the transparent plate as described above may not be used.
The substrate member may be a gel. Alternatively, in the case of a disposable biochip cartridge, a material that does not return after plastic deformation can be used as the substrate member.
[0047]
【The invention's effect】
As described above, the present invention has the following effects.
(1) Processing from sample injection to hybridization is performed consistently in a sealed cartridge. Therefore, accidents that come into contact with the injected solution due to an erroneous operation can be prevented.
(2) It is possible to easily produce an inexpensive biochip cartridge that can be easily manufactured with an inexpensive material.
[0048]
(3) Since the flow path is fixed, the remainder of the sample and uneven pressing are reduced, and accurate delivery is possible.
(4) A hard cover is used to press the substrate member, and autofluorescence from the cover is unlikely to occur.
[Brief description of the drawings]
FIG. 1 is a block diagram showing an embodiment of a biochip cartridge according to the present invention.
FIG. 2 is a diagram for explaining an operation;
FIG. 3 is a block diagram showing another embodiment of the present invention.
FIG. 4 is a block diagram showing still another embodiment of the present invention.
FIG. 5 is a block diagram showing still another embodiment of the present invention.
FIG. 6 is a block diagram showing still another embodiment of the present invention.
FIG. 7 is a block diagram showing still another embodiment of the present invention.
FIG. 8 is a block diagram showing an example of a conventional biochip.
9 is a plan view of the biochip of FIG. 8. FIG.
FIG. 10 is an explanatory diagram showing a conventional method of injecting a solution into a biochip.
FIG. 11 is a block diagram showing an example of another conventional biochip.
FIG. 12 is a block diagram showing an example of still another conventional biochip.
13 is an explanatory diagram showing a method for operating the biochip shown in FIG. 12. FIG.
[Explanation of symbols]
100 Biochip cartridge 101, 102 Cover 110 Substrate member 111 Channel 112 Collecting unit 113 First bag unit 114 Second bag unit 115 Pretreatment unit 116 Detection unit 117 Waste liquid container 118 Channel 200 Roller 210 Shutter 300 Fluorescence Reader

Claims (12)

A flat substrate member made of an elastic material;
A flexible cover having a material harder than the substrate member and attached in close contact with the upper and lower surfaces of the substrate member,
The substrate member includes a collection unit for storing biopolymers, a pretreatment unit for pretreating the biopolymers, and a biological height prepared in advance from the pretreated biopolymers. A detection unit for detecting a biopolymer that binds to a molecule, and each gap portion of a flow path connecting the collection unit, the pretreatment unit, and the detection unit,
A biochip cartridge characterized in that the biopolymer can be sequentially moved from the collection part side through the pretreatment part to the detection part side.   The movement of the biopolymer is performed by pressing the cover with a roller-like rigid body and crushing each gap portion of the substrate member from the collection unit to the detection unit side via the pretreatment unit. The biochip cartridge according to claim 1, wherein the cartridge is configured to be performed.   The substrate member is formed with a bag portion filled with a pretreatment liquid, and the pretreatment liquid stored in the bag portion is sent to the pretreatment portion when the roller is pressed. The cartridge for biochips according to claim 1 or 2.   The biochip cartridge according to any one of claims 1 to 3, wherein the substrate member is formed with a waste liquid storage section for storing the waste liquid discharged from the detection section.   The biochip cartridge according to any one of claims 1 to 4, wherein the cover is attached to a front surface and a back surface of the substrate member, respectively.   The biochip cartridge according to any one of claims 1 to 5, wherein the flow path formed in the substrate member is formed so that the gap is crushed by pressing the cover with the roller-shaped rigid body.   7. The biochip cartridge according to claim 1, wherein the cover is made of plastic, glass, or quartz.   8. The biochip cartridge according to claim 1, wherein the cover is made of a transparent material so that at least the detection unit can perform optical detection.   A plurality of bag portions for storing the pretreatment liquid are arranged at different positions, and when the substrate member is crushed by the roller-like rigid body, the pretreatment liquid flows from each bag portion into the pretreatment portion with a time difference. 9. The biochip cartridge according to claim 1, wherein the biochip cartridge is formed as described above.   The biochip cartridge according to any one of claims 1 to 9, wherein the substrate member is formed in a wedge shape whose thickness gradually decreases from the collection unit side toward the detection unit side.   The valve according to any one of claims 1 to 10, wherein a valve for stopping the flow of the solution is provided in the flow path, and the valve is opened when pressure is applied to the solution flowing through the flow path. Biochip cartridge.   12. The biochip cartridge according to claim 1, wherein the substrate member is made of a plastically deformable material or gel.

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