JPH06222035A - Detecting region carrying tape, detecting element, detector and detecting system - Google Patents

Abstract

PURPOSE:To provide a detecting region carrying tape, a detecting element, a detector and a detecting system. CONSTITUTION:The detecting region carrying tape 10 comprises a tape-like base 1 and a plurality of independent detecting regions 2 arranged thereon at an interval in the longitudinal direction wherein the detecting region 2 includes a porous reactant layer 3 carrying a biological catalyst, electrodes 4, 4', and joint terminals 5, 5'. The detecting element is an external reader jointing type containing a tape. The detector and the detecting system include detecting elements and a detecting region carrying tape. Since a pair electrode is provided on the disposable tape side and an electric signal is received on the repeatedly used device side, contamination is retarded while keeping high accuracy.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a tape in which a plurality of independent detection regions, each of which is composed of a porous material carrying an immobilized enzyme or the like and an electrode, are continuously arranged; The present invention relates to a joint type and disposable detection element; a detection device capable of inserting the detection element to detect an inspection object contained in blood, urine, etc .; and a detection system using the tape.

[0002]

2. Description of the Related Art Japanese Patent Laid-Open No. 63-165749 discloses
A biosensor having an electrode coated with a permselective membrane is described. In this biosensor, after attaching blood samples one after another to a fixed membrane tape carrying immobilized enzymes etc.,
The tape is continuously moved onto the selectively permeable membrane-coated electrode. The selective permeation membrane allows only the reaction product or reaction consumption product of the test trace substance in the sample and the enzyme in the fixed membrane to permeate, so that the change can be detected by the electrode. Therefore, in this biosensor, it is possible to always use a fresh one as the immobilization membrane that contacts the sample and performs the enzyme reaction. However, since the selective permeation membrane coated electrode that is in continuous contact with the fixed membrane is always the same, the selective permeation membrane and the electrode are contaminated with the sample, and there is a drawback that the detection accuracy is lowered. In order to maintain the detection accuracy, it is conceivable to frequently clean or replace the electrodes and the selectively permeable membrane, but the work becomes complicated and the cleaning effect is limited.

Further, in Japanese Patent Laid-Open No. 2-257050, a tape having sample permeation ports having selective permeation membranes arranged at predetermined intervals is used to measure the concentration of the tape so that the permeation ports can move on the immobilized enzyme electrode. The device is described. In this device, since the permselective membrane is moved, this portion is not contaminated, but since the immobilized enzyme electrode is repeatedly used, it is contaminated and it is difficult to maintain the detection accuracy.

[0004]

DISCLOSURE OF THE INVENTION The present inventor has conducted various studies to solve the above-mentioned drawbacks of the prior art.
Repeated use by providing a simple electrode pair on the side of the immobilized enzyme-carrying tape that is used only once and used only once, and the device side that is used repeatedly receives only the signal from the electrode It has been found that the device to be used is not contaminated by the sample, the detection accuracy can be maintained at a high level, and there is no need to use a permselective membrane. The present invention is based on these findings.

[0005]

SUMMARY OF THE INVENTION Therefore, the present invention provides
(A) a tape-shaped base, and (B) a plurality of independent detection regions arranged on the base at intervals in the longitudinal direction,
Each of the independent detection regions described above (1) is a porous support carrying a biological catalyst capable of reacting with a test object contained in a sample to cause a change detectable by an electrode. Reactant layer, (2) A pair of electrodes that are in contact with the porous reactant layer and that can detect changes due to the reaction between the test object contained in the sample and the biological catalyst,
And (3) a detection area-carrying tape, comprising a joining terminal capable of sending a signal from the electrode to an external reader. Further, the present invention provides (1) a storage portion for the detection area supporting tape, (2) a sample receiving port, and (3).
External reading device joining part, (4) connecting part with means for giving transfer driving force to the detection area carrying tape, (5) accommodating part of used detection area carrying tape, and (6) transfer driving force Also relates to a detection element, characterized in that it includes means for sequentially guiding the detection area carrying tape to the storage section, the sample receiving port, the external reader joining section and the storage section. Furthermore, the present invention further comprises (1) a mounting portion for the detection element, (2) a sampling device for communicating with a sample inlet of the detection element, (3) communication for an external reader joint portion of the detection element, The present invention also relates to a device for reading a signal from a joining terminal of the detection area supporting tape, and (4) a detection device including means for applying a transfer driving force to the detection area supporting tape. Furthermore, the present invention relates to a detection system using the above-mentioned detection area-supporting tape, and the detection system as a whole includes (1) a storage portion for the detection area-supporting tape, (2) a sampling device, and (3). A means for supplying the sample collected by the sample collecting device to the porous reactant layer of the detection area supporting tape, (4) an electric signal reader, (5) an electrode in contact with a joining terminal of the detection area supporting tape. Means for sending the signal to the electric signal reader, (6) means for applying a transfer driving force to the detection area carrying tape, (7) means for accommodating the tested detection area carrying tape, and (8)
A means for sequentially guiding the detection area carrying tape from the storage portion to the storage portion by the transfer driving force,
The detection system includes (A) a tape storage unit that stores at least the detection area supporting tape, and (B) a reading unit that includes at least the sampling device and an electric signal reading device. It also relates to said detection system.

A typical mode of the detection area supporting tape according to the present invention is shown in FIGS. 1 and 2 (FIG. 2 is a sectional view taken along the line AA of FIG. 1). The detection area supporting tape 10 comprises a tape-shaped substrate 1 and an independent detection area 2 formed thereon.
A plurality of independent detection areas 2 are arranged discontinuously and independently at intervals in the longitudinal direction of the tape-shaped substrate 1. Each independent detection region 2 includes a porous reactant layer 3, a pair of electrodes 4, 4 ', and bonding terminals 5, 5'. In the detection area supporting tape 10 according to the present invention, a buffer area 6 can be provided between the independent detection areas 2 as required. In this case, a barrier section 7 for preventing the permeation of the liquid sample is provided between the independent detection area 2 and the buffer area 6.
It is preferable to avoid the influence between the independent detection areas 2. Further, the insulating layer 8 is provided between the porous reactant layer 3 and the joining terminal 5. If necessary, the pitch guide holes 9 may be provided in both ears of the tape-shaped base 1.

The tape-shaped substrate 1 is made of a sheet-shaped material such as an electrically insulating and moisture-resistant polyester film or polyolefin film. The electrode 4 can be provided on the substrate 1 by printing a paste such as silver or carbon or by plating a printed circuit made of copper or the like with a metal such as platinum. It is preferable to simultaneously form the electrode pairs 4, 4 ′ and the joining terminal 5 because the manufacturing process is simplified.
At least one portion of the electrode pair 4, 4 ′ and the porous reactant layer 3 may be in contact with each other. The material of the porous support forming the porous reactant layer 3 is not particularly limited, but a fiber aggregate (for example, non-woven fabric, felt, woven fabric, knitted fabric) or foam or a composite thereof is preferable. The nonwoven fabric is particularly preferable because it has a three-dimensional three-dimensional structure and has a good liquid absorbing property. Although not shown, a separation filter such as a membrane filter or a filter paper is provided on the porous reactant layer 3 for the purpose of removing contaminants contained in the test object or particles interfering with the reaction. Good.
The joining terminals 5, 5 ′ are in electrical communication with the electrode pairs 4, 4 ′, but are electrically insulated from the porous reactant layer 3 by the insulating layer 8. The insulating layer 8 continuously or discontinuously in the longitudinal direction on the tape-shaped substrate 1 separates a part of each electrode pair 4, 4 ′ from the joining terminals 5, 5 ′ and the electrode pair 4, 4 ′. Thus, the porous reactant layer 3 is preferably laminated on the electrode pair 4, 4 ′ which is not covered by the insulating layer 8.

The buffer region 6 may be formed separately from the porous reactant layer 3, but the porous reactant layer 3 is continuously formed on the tape-shaped substrate 1 and a partial region thereof is formed in the buffer region 6. Is preferable because the manufacturing process is simple. Buffer area 6
Not only prevents the liquid sample from moving between the independent detection regions 2, but also has a cleaning action of wiping off the liquid sample supplied to the preceding independent detection region 2 when it adheres to the detection device. The barrier portion 7 can be formed, for example, by heat-sealing the porous reactant layer 3. Therefore, according to a preferred embodiment of the detection area supporting tape 10, the porous reactant layer 3 continuously provided in the longitudinal direction on the tape-shaped substrate 1 is provided by the barrier portion 7 composed of a heat seal or the like in each predetermined area. The independent detection areas 2 and the buffer areas 6 are alternately and continuously arranged by partitioning, and the area including the electrode pairs 4 and 4'is used as the independent detection area 2.
A region that does not include the electrode pairs 4 and 4 ′ is used as the buffer region 6. It is not necessary to provide the pitch guide hole 9 when the tape is intermittently transferred at a constant distance using a pulse motor or the like.

Although not shown in FIGS. 1 and 2,
To prevent the porous reactant layer from contacting with gas or liquid during the storage period from the preparation of the detection area-carrying tape of the present invention to its actual use, resulting in contamination or deterioration of the detection function. In addition, it is preferable to provide a protective film that can be easily removed on the porous reactant layer. In this case, immediately before using the detection area supporting tape of the present invention, the protective film is removed to expose the porous reactant layer. The protective film may be discontinuously provided only on the porous reactant layer. For example, when the protective film is provided on the detection area supporting tape shown in FIG. 1 and FIG. It is preferable to provide in the form of a peeling tape that continuously covers the entire area of the porous reactant layer supporting surface or the entire area other than the pitch guide holes 9 because the arrangement and the peeling operation are simple.
When preparing this continuous peelable protective film tape using an adhesive, the porous reactant layer or the like carried on the detection area carrying tape, when peeling the protective film tape It should not be peeled off together with the protective film tape. As the protective film, a synthetic resin film having gas and / or liquid barrier properties,
For example, polyvinyl chloride resin, polyester resin,
Alternatively, it is made of polyolefin resin and has a thickness of about 10-1.
A 00 μm film can be used.

The porous reactant layer 3 may be continuous in the longitudinal direction of the detection area supporting tape 10 as described above, but may be formed discontinuously in some cases.
As shown in FIG. 3, a small piece of the porous reactant layer 3 may be stuck on each electrode pair 4, 4 ′. In the three independent detection areas 2 (A, B, C) shown in FIG. 3, A is an insulating layer 8 provided on the pair of electrodes 4, 4'and the joining terminals 5, 5 ', and a circular small piece is further provided thereon. Shows the state of the complete independent detection region provided with the porous reactant layer 3, and B shows the state of the complete independent detection region from the porous reactant layer 3 and the insulating layer 8.
Is removed to show the modes of the electrodes 4, 4'and the bonding terminals 5, 5 ', and C is a part of the porous reactant layer 3 cut away. It is preferable that all of the three independent detection areas 2 (A, B, C) be protected by a continuous peeling type protective film 37 until just before use.

In the case of the embodiment shown in FIG. 3, the porous reactant layer 3
Must be cut into small pieces of a predetermined shape and attached to predetermined positions on each electrode pair, but on the other hand, there is no portion such as a buffer area that does not contribute to the reaction of the porous reactant layer 3. , The biological catalyst is effectively used without waste. Also,
Since the porous support does not exist between the small pieces of the porous reactant layer 3, the sample solution is adjacent to the porous reactant layer 3
There is no need to provide a barrier because there is no risk of moving to small pieces. Further, if a small piece of the porous support is adhered on each electrode pair, then the biological catalyst is applied in the form of a solution and dried to form the porous reactant layer 3, This is because the amount of the biological catalyst in the independent detection region can be accurately controlled, and the porous reactant layer 3 in which the biological catalyst is uniformly dispersed can be obtained.

A biological catalyst is supported on the entire surface of the porous support constituting the porous reactant layer 3 used in the independent detection area 2 of the detection area supporting tape 10 of the present invention.
The biological catalyst may be any as long as it can react with an object to be tested contained in the sample to cause a change detectable by the electrode 4, and examples thereof include an enzyme, an antibody, an organelle,
Microorganisms or animal and plant cells can be used. In the present invention, the biological catalyst is selected according to the type of the test object. As the biological catalyst, the same one may be provided for each independent detection region 2, or a different one may be provided for each independent detection region 2 or for each group of independent detection regions 2. It is preferable to select a combination that finally produces hydrogen peroxide or ammonia by the reaction of the test object with one or more biological catalysts (particularly enzymes). Although many examples are already known as combinations thereof, typical examples are glucose-glucose oxidase (hydrogen peroxide), pyruvate-pyruvate oxidase (hydrogen peroxide), and urate-uricase (peroxidation). Hydrogen), urea-urease (ammonia), neutral fat-lipase and glycerol oxidase (hydrogen peroxide), cholesterol-cholesterol oxidase (hydrogen peroxide), L amino acid-L amino acid oxidase (hydrogen peroxide), ethanol-alcohol oxidase (Hydrogen peroxide), antibody protein-antibody and peroxidase. Further, the electrode is selected according to the type of the reaction product of the test object and the biological catalyst. For example, with respect to the hydrogen peroxide generation reaction, the reaction can be detected by a potentiostatic electrolysis method using electrodes of platinum, gold, silver, carbon and the like. Further, an ammonia electrode or an ion-selective field effect transistor (FET) electrode can be preferably used for the ammonia production reaction.

As a method for supporting the biological catalyst on the porous support, any method can be used,
For example, a carrier binding method in which a biological catalyst is directly or indirectly bound to the surface of a porous support by a covalent bond or an ionic bond (for example, the method described in JP-A-60-224618), silk fibroin, etc. Use the encapsulation method (the method described in JP-A-2-245189 or JP-A-60-120988) in which the biological catalyst is confined and immobilized in the lattice of the polymer gel or in the resin film. You can A method of attaching a biological catalyst to a porous support together with a surfactant and a stabilizer is also useful.

Various detection elements can be constructed using the detection area supporting tape 10 of the present invention. A typical mode thereof is shown in FIGS. 4 and 5. The audio cassette tape type detection element 20 includes a housing 21, a housing 22 for the detection area supporting tape 10, a sample receiving port 23, an external reading device joining portion 24, a connecting portion 25 for connecting a pitch feed guide roll and a pulse motor. It includes an accommodating portion 26 for the used detection area carrying tape, and a guide means 27 for the detection area carrying tape. Further, when the detection area supporting tape 10 carries the protective film tape 37, the guide means 38 of the protective film tape 37 after peeling and the accommodating portion thereof (in the illustrated mode, also serve as the accommodating portion 26 of the detection area supporting tape). )
including.

The detection area carrying tape 10 wound around the reel 28 is separated from the protective film tape 37, and then the roller 27 is rotated by the intermittent rotation of the pitch feed guide roll 29 which meshes with the pitch guide hole 9. After that, the sample is transferred to the sample receiving port 23 and temporarily stopped. At the time of this temporary stop, the sample is dropped from the sample supply nozzle 30 onto the porous reactant layer 3 in the independent detection region 2. Sample inlet 23
It is preferable to provide the support base 31 on the inside. Then, the pitch feed guide roll 29 is rotated to transfer the independent detection region 2 on which the sample has been dropped to the external reading device joining portion 24 and temporarily stop. Here, the detection area supporting tape 1
The connection terminals 5, 5 ′ of 0 are brought into contact with the connection terminals 34, 34 ′ provided at the tip of the reading head 33 which communicates with the external reading device 32. It is preferable to provide the support base 35 also inside the joint portion 24. After inspection, pitch feed guide roll 2
9 is rotated to attach the detection area carrying tape 10 to the roller 27.
After that, the reel 36 is wound up.

On the other hand, the protective film tape 37 is wound around the reel 36 via the roller 38. The take-up reel of the protective film tape 37 is the used detection area carrying tape 10.
Although it may be provided separately from the take-up reel 36, the number of parts can be reduced and the space can be reduced by using both of them. The rotation and stop of the pitch feed guide roll 29, the dropping of the sample by the sample supply nozzle 30, and the reading head 3
The contact between 3 and the contact terminals 5, 5'is tuned by a suitable control device.

In the embodiment shown in FIGS. 4 and 5, the connecting portion 25 for the pitch feed guide roll and the pulse motor is provided behind the external reader joining portion 24, but before the sample receiving port 23,
It may be provided between the sample receiving port 23 and the external reader joining section 24. The external reading device joining section 24 does not necessarily have to be provided on the detection area supporting tape 10 as shown in FIG. 5, and may be provided near the sample receiving port 23, for example. In this case, at the position where the sample is supplied at the sample receiving port 23, the connecting terminals 5 and 5'of the detection area carrying tape are provided.
The electric signal transmitting means that can contact with the casing 21 may be provided in the casing 21. It is desirable that the electric signal transmitting means is provided with connection terminals at both ends of an electrically conductive material, whereby one end portion is connected to the connection terminals 5 and 5'of the detection area carrying tape and the other end portion is connected. The external reader connection portion 24 can be brought into contact with the connection terminals 34, 34 ′ provided at the tip of the read head 33. With this configuration, by applying a potential between the electrode pairs 4 and 4'in advance, the sample can be attached to the porous reactant layer 3 and the reaction can be detected at the same time, and the sample can be detected from the sample receiving port 23 to the outside. Reader junction 24
It is possible to eliminate the detection error due to the transfer of the tape to the. A window 39 made of a transparent material is preferably provided above the reels 28 and 36 so that the remaining amount of the unused detection area carrying tape 10 can be confirmed. Further, as the external reading device 32, a potentiostatic electrolysis device or the like can be preferably used, and it is preferable to provide an appropriate recording and / or display device in communication therewith.

The detection element shown in FIG. 4 is connected to the installation portion of the detection element, the sampling device which communicates with the sample inlet of the detection element, and the external reader connection portion of the detection element, It can be used in combination with a detection device including a device for reading a signal from the joining terminal of the detection area supporting tape and a means for applying a transfer driving force to the detection area supporting tape. This detecting device can be used for urine inspection without touching the urine sample by mounting the sample collecting device in the toilet bowl in a centralized laboratory such as a hospital.

The present invention also relates to various detection systems using the detection area supporting tape. This detection system includes (A) a disposable tape storage unit that stores the detection area supporting tape and is used only for one detection, and (B) a reading unit that includes at least a sampling device and an electric signal reading device. When the configuration of the tape storage unit (A) is changed, the configuration of the other reading unit (B) is also changed. The tape storage unit (A) having the simplest structure stores the detection area supporting tape in a roll shape in the cylindrical casing, and the tip of the detection area supporting tape can be taken out from the slit provided on the side surface of the cylindrical casing. , Roll photographic film of Patrone type. In this case,
In the reading unit (B), in addition to the sampling device and the electric signal reading device, means for supplying a sample collected by the sampling device to the porous reactant layer of the detection area supporting tape, the detection A means for sending a signal from the electrode to the electric signal reading device in contact with the joining terminal of the area carrying tape, a means for applying a transfer driving force to the detection area carrying tape, a means for accommodating the tested detection area carrying tape, or A means for separating and discarding, and a means for guiding the detection area carrying tape from the accommodating section to the accommodating section in order by the transfer driving force are arranged. It is preferable to collect the used detection area carrying tapes, preferably in a bag provided in the accommodating section, and discard the whole bag. On the other hand, one example in which the tape storage unit (A) is provided with various means is the detection element 20 shown in FIGS. 4 and 5. Therefore, there are various modifications between the roll photographic film cartridge type tape storage unit (A) and the detection element 20 type tape storage unit (A).
The configuration of the other reading unit (B) also changes accordingly.

The detection device or the detection system of the present invention can be used for detecting and quantifying various biological test objects contained in an arbitrary sample, and in particular, a biological liquid sample,
For example, lipids (eg, cholesterol, phospholipids, neutral fats) contained in blood, serum, plasma, etc., sugars or nitrogen compounds (eg, urea, uric acid, creatinine), hormones (eg, growth hormone, luteinizing hormone, etc.) It is used for detection and quantification of adrenocorticotropic hormone, insulin, gastrin), various marker proteins (for example, tumor markers), sugars contained in urine and the like, and nitrogen compounds (for example, uric acid and urea nitrogen). Prior to performing the test with the detection element of the present invention, the biological liquid sample is optionally diluted with physiological saline or the like, particles such as blood cells are separated and removed, or treated with a drug such as an anticoagulant. You can also do it.

[0021]

The detection area supporting tape of the present invention has a simple electrode pair carried together on the side of the immobilized enzyme carrying tape which is used only for one measurement and is disposable, and repeatedly used in combination therewith. By adopting a configuration in which only the electric signal from the electrode is received on the device side, the device side that is repeatedly used is less likely to be contaminated by the sample, the detection accuracy can be maintained at a high level, and it is necessary to use a selective permeation membrane etc. Nor. Further, by using the detection element of the present invention, continuous inspection can be easily performed without directly touching the sample. Further, in the present invention, various detection systems can be configured by using the above-mentioned detection area supporting tape.

[Brief description of drawings]

FIG. 1 is a partial plan view showing a notch of a mode of a detection area supporting tape according to the present invention.

FIG. 2 is a sectional view taken along the line AA of FIG.

FIG. 3 is a partial plan view showing a notch of another embodiment of the detection area supporting tape according to the present invention.

FIG. 4 is a perspective view of an embodiment of a detection element according to the present invention.

FIG. 5 is a cross-sectional view schematically showing a state in which the detection element of FIG. 4 is incorporated into a detection device of the present invention and used.

[Explanation of symbols]

1 ... Tape-shaped substrate; 2 ... Independent detection area; 3 ... Porous reactant layer; 4 ... Electrode; 5 ... Joining terminal; 10 ... Detection area carrying tape; 20 ... Detection element; 22 ...・ Accommodation area for the detection area supporting tape; 23 ・ ・ Sample inlet; 24 ・ ・
External reading device splicing part; 25 ... Pitch feed guide roll contact part; 26 ... Used detection area carrying tape accommodating part; 2
7 ... Guide means for the detection area supporting tape; 30 ... Sample supply nozzle; 32 ... Reader; 33 ... Read head; 3
7. Protective film.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification code Internal reference number FI technical display location G01N 27/327 27/416

Claims (4)

[Claims] 1. A tape-shaped substrate (A), and (B) a plurality of independent detection regions arranged on the substrate at intervals in the longitudinal direction. Each of the independent detection regions is (1) a sample. A porous reactant layer carrying a biological catalyst capable of reacting with a test object contained therein to cause a change detectable by an electrode, (2) the porous reaction A pair of electrodes that are in contact with the body layer and can detect changes due to the reaction between the test object contained in the sample and the biological catalyst; and (3) sending a signal from the electrodes to an external reader. A detection area-carrying tape, comprising: 2. A storage unit for the detection area supporting tape according to claim 1, (2) a sample inlet, (3) an external reader joining section, (4) a transfer driving force for the detection area supporting tape. To the means for supplying the detection area carrying tape, (5) a housing portion for the used detection area carrying tape, and (6) the detection area carrying tape by the transfer driving force, the housing portion, the sample receiving port, and the outside. A detection element, characterized in that it comprises means for guiding in sequence to the reader joint and the housing. 3. A detection element installation part according to claim 1, (2) a sampling device which communicates with a sample inlet of the detection element, and (3) a junction part of an external reader of the detection element. A detection device, comprising: a device for reading a signal from a joining terminal of the detection area supporting tape according to claim 1; and (4) means for applying a transfer driving force to the detection area supporting tape. 4. A detection system using the detection area carrying tape according to claim 1, wherein the detection system as a whole comprises (1) a storage area for the detection area carrying tape according to claim 1, and (2) a sampling device. (3) means for supplying the sample collected by the sample collecting device to the porous reactant layer of the detection area supporting tape,
(4) electrical signal reader, (5) means for sending a signal from an electrode to the electrical signal reader by contacting a joining terminal of the detection area carrying tape, (6) transfer driving force for the detection area carrying tape And (7) means for accommodating the inspected detection area supporting tape, and (8) means for sequentially guiding the detection area supporting tape from the accommodating portion to the accommodating portion by the transfer driving force. The detection system is composed of (A) a tape storage unit for storing at least the detection area supporting tape, and (B) a reading unit including at least the sampling device and an electric signal reading device. , The detection system described above.