JP4670013B2 - Biosensor and manufacturing method thereof - Google Patents

Description

The present invention relates to a biosensor and a method for producing the same. More specifically, the present invention relates to a biosensor having a resist layer covering the surface of a conductor constituting electrodes and terminals and a method for producing a biosensor having a folding step.

Conventionally, disposable sensors (Patent Document 1 and Patent Document 3) have a three-dimensional structure in order to ensure quantification, and further, a sample solution is automatically obtained by utilizing a capillary phenomenon (Patent Documents 5 and 6). In particular, a mechanism for introducing the sensor inside the sensor is known (Patent Document 7). The sensor having such a structure is assembled by stacking a spacer and a cover on an electrically insulating substrate. An electrode pattern is formed on the substrate, and air holes necessary for air necessary for capillary action to escape are formed on the cover. Each of these components must be punched into a predetermined shape in advance, and positioning for accurate overlaying of each component in 3D processing is also required. Becomes complicated. Furthermore, when these sensors require application of reagents such as molecular identification elements and mediators (Patent Documents 2 and 4) and formation of films (Patent Document 8) to avoid the influence of interfering substances Has a problem that it becomes a more complicated process.
JP 47-500 A JP-A-48-37187 JP-A-52-142585 JP-A-54-50396 JP 56-79242 A JP-T 61-502419 Japanese Patent Laid-Open No. 1-291153 Japanese Patent Laid-Open No. 3-202864

  The conventional sensor described above requires many processes and materials for manufacturing, and has to take a complicated structure. As a result, a large capital investment was required for the production line, the product yield was not sufficient, and the cost was high. Naturally, the environmental load at the time of material procurement and manufacturing was also large. Furthermore, due to complicated processes, especially alignment during substrate lamination, the coefficient of variation (CV), which is an indicator of variations in sensor characteristics produced, was not sufficient. In addition, since the change in shape of the biosensor causes a decrease in measurement accuracy and reproducibility, it has been required to ensure long-term shape stability in the biosensor without causing warping of a cover or the like after manufacturing. .

In order to solve the above problems, the inventors have previously proposed a biosensor manufactured by folding, bending, or bending a single electrically insulating flat substrate. In this biosensor, an electrode is formed on a single electrically insulating substrate, and the electrode arrangement is made flat or three-dimensional by processing a single flat substrate in three dimensions so that the electrode is arranged inside the substrate. Quantitative measurement in a narrow region is possible, and the main structure of the sensor is formed from a single flat substrate. Accordingly, in order to prevent the folded portion from being warped, it is necessary to attach a fixing tool to the folded portion, thermocompression bonding, cutting, etc., and to make a space for the spacer formed between the substrate and the cover. Since the sample inlet is formed by using the sample solution, the sample liquid permeates into the groove formed at the boundary between the substrate near the sample inlet, the spacer, and the constituent materials of the cover, and there is a problem that the sample volume fluctuates. .
Japanese Patent Laid-Open No. 2005-233917

The problem of the biosensor will be described in detail with reference to FIG. a) and b) differ only in the shape of the substrate 1, and show an assembly example of a conventional biosensor. In i), conductors 7 and 7 are formed on the surface, and the surface of the conductor other than the one substrate 1 provided with the perforations 16 to be folded and the electrode reaction unit 13 and the terminal 11 is provided. An overlying resist layer 6 is shown. The resist layer 6 also functions as the spacer 2. ii) shows a substrate 1 having a resist layer 6 formed on the surface and an adhesive layer 5 to be coated in the next assembly step. Here, the adhesive layer 5 also functions as the spacer 2 in the same manner as the resist layer 6. In iii), the substrate on which the adhesive layer 5 is formed is folded along the perforations 16 and shows a state before being overlapped. In iv), the biosensor 3 which is the folded molded body 14 formed by the substrate 1 is shown. In this case, since the folded portion formed along the perforation 16 may be warped depending on the thickness of the spacer such as the resist layer 6 and the adhesive layer 5, a fixing tool is attached to this portion or the warping stress is caused by thermocompression bonding. Some kind of treatment such as removing was necessary.

  FIG. 17 is a diagram illustrating a usage example of the biosensor illustrated in FIG. 16. i) shows a state before the sample solution 17 is introduced into the sample introduction port 9 of the biosensor 3, ii) shows a state during introduction, and iii) shows a state after introduction. Here, as shown in iii), the state in which the sample liquid 17 is dyed on the side surface of the sensor 3 near the sample introduction port 9 is shown.

  This state is shown more clearly in FIG. 18 which shows an example of use in the cross-sectional view of the biosensor shown in FIG. 16b) iv). Regarding the AA ′ cross section (part removed from the electrode reaction part 13) and the BB ′ cross section (part of the electrode reaction part 13) of the sensor, i) is the state before introducing the sample liquid 17, and ii) is the sample liquid. 17 shows the state after introduction. As is apparent from ii), in the case of the conventional sensor, the sample liquid is shown to wrap around the lower side of the sensor 3, that is, the side surface facing the sample introduction port 9 in addition to the sample transport path 8.

  As described above, such a foldable sensor has succeeded in greatly improving the manufacturing method of the conventional sensor by greatly simplifying the manufacturing process, reducing the material, and extremely simple structure. In order to prevent the folding part from warping, it is necessary to attach a fixing tool to the folding part, thermocompression bonding, cutting, etc., and there is no spacer formed between the substrate and the cover Since the sample introduction port is formed by using a space, the sample liquid may wrap around the sample introduction port provided in the empty part of the spacer sandwiched between the two substrates. There was a problem that fluctuated. The change in the sample volume may result in a change in the measured value as a result, and there is currently a demand for some measures to prevent the wraparound.

  An object of the present invention is a biosensor that can be manufactured without requiring many processes and materials as in the case of a conventional sensor, and the manufactured biosensor does not cause a shape change, and further, a sample volume. It is an object of the present invention to provide a biosensor and a method for producing the same.

An object of the present invention is to provide a biosensor in which electrodes, spacers, and a cover are sequentially formed on an insulating substrate. The insulating substrate and the cover are connected by a soft sheet, and a part of the soft sheet constitutes an electrode and a terminal. It acts as a resist layer covering the surface of the conductor other than the part to be covered , and the resist layer forms a part of the spacer and is folded at the soft sheet portion connecting the insulating substrate and the cover, and the electrode is housed inside the biosensor. The biosensor is formed by sequentially forming a soft sheet layer and an adhesive layer on the surface of the insulating substrate and the cover on which the electrode is formed, and then insulating the electrode so that the electrode is placed inside the biosensor. Manufactured by folding the insulating substrate and cover at the soft sheet part connecting the insulating substrate and cover It is.

In the biosensor according to the present invention, since the insulating substrate and the cover are connected by a soft sheet , a part of which acts as a resist layer , the folded portion does not warp, and the folded portion to prevent this occurs. Excellent effects such as no need for mounting fixtures, thermocompression bonding, and cutting.

In addition, when the sample inlet for the sample to be measured is provided in the soft sheet part that connects the insulating substrate and the cover, it is possible to prevent the sample liquid from getting around the sample inlet. It becomes possible to specify.

Further, the needle-integrated biosensor according to the present invention has a puncture needle that breaks through a soft sheet portion that connects the insulating substrate and the cover during blood collection when the puncture needle is included and fixed in the biosensor having a folding structure. The puncture needle returns to its original position by the restoring force of the soft sheet material, and blood collection is introduced from the newly formed sample introduction port at that time, so that the blood collection component can be measured electrochemically.

  As the substrate and the cover, it is sufficient if they are electrically insulating, for example, plastic, biodegradable material, paper or the like is used, and preferably polyethylene terephthalate is used. Further, an oxygen permeable material can be used, and in this case, since the reagent can be prevented from being reduced, there is an effect of suppressing variation in the measured value depending on the state of reduction.

Further, for the cover, the same material as the soft sheet forming material described later can be used. In this case, the cover and the resist layer are configured so as to be integrated with each other, so that the soft sheet layer becomes the insulating substrate and the cover. It can be set as the aspect which connects . Thereby, a cover member is not required separately and the structure of a biosensor can be simplified more.

  The electrode is formed on the substrate by a screen printing method, a vapor deposition method, a sputtering method, a foil pasting method, a plating method, etc., and the materials include carbon, silver, silver / silver chloride, platinum, gold, nickel, copper, Examples include palladium, titanium, iridium, lead, tin oxide, and platinum black. Here, as the carbon, carbon nanotubes, carbon microcoils, carbon nanohorns, fullerenes, dendrimers, or derivatives thereof can be used.

  The electrode may be a two-pole method formed with a working electrode and a counter electrode or a three-pole method formed with a working electrode and a counter electrode, a reference electrode, or an electrode method with more poles. Here, when the tripolar method is adopted, in addition to the electrochemical measurement of the measurement target substance, it is possible to measure the moving speed of the blood sample introduced into the transport path, thereby measuring the hematocrit value. Moreover, you may be comprised by 2 or more sets of electrode systems.

  A reagent layer (electrode reaction part) can be formed on the substrate on which the electrode is formed. The reagent layer is formed by a screen printing method or a dispenser method, and the reagent layer can be immobilized on the electrode surface or the substrate surface by an adsorption method involving drying or a covalent bonding method. Examples of the reagent arranged in the electrode reaction part of the biosensor include those containing glucose oxidase as an oxidase and potassium ferricyanide as a mediator when configured for blood glucose measurement. When the reagent is dissolved by the blood, the enzyme reaction is started. As a result, potassium ferricyanide coexisting in the reaction layer is reduced and potassium ferrocyanide, which is a reduced electron carrier, is accumulated. The amount is proportional to the substrate concentration, ie the glucose concentration in the blood. The reduced electron carrier accumulated for a certain time is oxidized by an electrochemical reaction. An electronic circuit in the main body of the measuring apparatus, which will be described later, calculates and determines the glucose concentration (blood glucose level) from the anode current measured at this time, and displays it on the display unit arranged on the main body surface.

  In addition, a surfactant and lipid can be applied around the blood collection port and on the surface of the electrode or reagent layer (electrode reaction part). By applying a surfactant or lipid, the sample can be moved smoothly.

  In the biosensor in which the reagent layer is provided on the electrode filled with the above blood collection, the blood collection and the reagent react when the blood collected from the blood collection port contacts the reagent layer on the electrode. This reaction is monitored as an electrical change at the electrode.

(Reagent layer formation) The area of the electrode is defined by a resist layer formed by a part of the soft sheet . The soft sheet is made of a soft material that does not react or dissolve with the substrate and does not cause bending of the folded portion when folded, such as ultraviolet or visible light curable vinyl / acrylic resin, urethane acrylate resin, polyester. It is made of an acrylate resin, polyvinyl chloride, polyethylene, polyester, polyolefin, polyvinyl fluoride, cellophane, etc., and has a thickness of about 5 to 500 μm, preferably about 10 to 100 μm. The purpose of the use of the resist layer covers the conductor surface other than portions constituting the electrodes and the terminals, in addition to defining the electrode area by clarifying the electrode patterns, to insulate the sample transfer path does not exist reagent layer There is a purpose . The soft sheet including the resist layer as a part has an important role of connecting the insulating substrate and the cover in addition to the purpose of the resist layer . By providing a sample introduction port in a part of the soft sheet part connecting the insulating substrate and the cover, it is possible to prevent a sample liquid from flowing around the sample introduction port and to manufacture a sensor that accurately defines the sample volume. Has an excellent effect. In addition, the soft sheet can be used to prevent contact with the reagent layer arranged in the electrode reaction layer in a needle-integrated biosensor provided with a puncture needle, which will be described later, and also used as a puncture membrane during puncture blood collection You can also When used as a puncture membrane, it also functions as a sample introduction port due to a through hole formed after puncture.

The soft sheet layer described above can be formed by (1) a screen printing method, (2) a thermocompression laminating method, or (3) a method using an adhesive tape. In particular,
(1) After disposing a release plate between the insulating substrate and the cover, a soft sheet layer is formed by a screen printing method. Examples of the material of the release plate include silicone, tetrafluoroethylene, polyethylene, and the like. A material obtained by coating these release materials on the surface of an arbitrary plate material can also be used. The release plate is removed by peeling when the soft sheet portion between the insulating substrate and the cover is folded.
(2) Connect the two insulating substrates using adhesive tape such as cellophane tape. However, since the adhesive portion is exposed between the two insulating substrates, it is preferable to cut the exposed portion after forming the sensor.
(3) The pressure-bonding surface of the polypropylene sheet capable of thermocompression bonding is stacked on the electrode forming substrate and thermocompression bonded under the conditions of 100 ° C. and 250 kg / cm ² . At this time, it is sufficient to add the thermocompression bonding for the portion that needs to maintain the flexibility of the sheet. In such a thermocompression- bondable sheet , a flexible sheet layer is formed by a method in which a commercially available laminate material such as polyvinyl chloride, polyethylene, polyester, polyolefin, and polyvinyl fluoride can be used as it is. The resist layer formed by a part of such a soft sheet may contain at least one reagent among enzymes, mediators, and surfactants.
Japanese Patent Laid-Open No. 56-42652

The substrate and the cover on which the soft sheet layer is formed are bonded via an adhesive such as an acrylic resin adhesive to constitute a biosensor. Such an adhesive layer can also be formed by a screen printing method, and is formed with a thickness of about 5 to 500 μm, preferably about 10 to 100 μm, and such an adhesive layer acts as a spacer as well as a resist layer. In addition, the said reagent can also be contained in an adhesive bond layer.

In addition, the biosensor having the above configuration is provided with a soft sheet layer so as to connect a long substrate having a large number of electrodes and a long cover, and then folded along a portion connecting the substrate and cover of the soft sheet . Later, a large number of biosensors can be manufactured at once by punching into a sensor shape. The needle-integrated biosensor manufactured by such a manufacturing method has very good reproducibility and has features that cannot be achieved by a conventional lamination method.

  A puncture needle for collecting body fluid from the skin of the subject can also be disposed on the adhesive layer. As the puncture needle, it is necessary to puncture the subject, and it is desirable that the puncture needle is strong and sharp enough to withstand this, and a thin puncture needle is preferable in order to suppress pain during puncture. Specifically, a 21-33 gauge thing by Terumo company is used. The puncture needle may be a hollow needle or a rod-like needle as long as it can penetrate the subject's skin. Furthermore, since the puncture needle needs to be hygienicly stored in the biosensor until it is used, a photocatalytic function effective for antibacterial and antiviral effects may be imparted to the needle tip surface. In that case, a film of titanium oxide or titanium dioxide is desirable.

  Here, there is a possibility that the puncture needle contained in the inside of the sample conveyance path is contaminated by the application of the reagent layer, the surfactant or the lipid into the sample transport path. In order to prevent such contamination, it is preferable not to apply these reagents around the tip of the puncture needle, or to isolate them from the reagent layer in the sample transport path by a resist layer or an adhesive layer.

The needle-integrated biosensor having the above-described configuration, in which the inside of the biosensor is sealed, is manufactured under a negative pressure condition, preferably a vacuum condition, than the outside air, so that the sensor is in a negative pressure state. In addition to capillary action, the suction means can be used in combination for the movement of blood collection into the sample transport path after puncture. By adopting such a configuration, blood can be collected smoothly. Here, a blood collection introduction guide can be provided in the vicinity of the puncture blood collection port. Examples of the material for the blood collection introduction guide include gels, elastic materials, and foamable materials, and the same material as the soft sheet can be used. A blood collection introduction guide made of such a material maintains the negative pressure and also has the effect of improving the adhesion between the skin of the subject and the puncture blood collection port.

In the needle-integrated biosensor of the present invention, it is desirable that a series of operations of puncture, blood collection, and measurement be performed by a measurement device having a puncture drive. In this case, for example, with respect to puncture driving, it is desirable to have a mechanism in which the needle penetrates the soft sheet of the biosensor and breaks through the skin of the subject, and a mechanism that quickly returns to the original position immediately after puncturing.

  An example of the structural features of the measuring device will be described. In this measurement apparatus, the puncture needle drive unit and the measurement apparatus unit are integrated, and the puncture needle drive unit includes a trigger unit, a puncture start button unit, and a drive unit using an elastic body such as a spring. On the other hand, for the measuring device section, the sensor introduction section, the connector, the electrochemical measurement circuit, the memory section, the operation panel, the measurement section that measures the electrical values at the electrodes of the biosensor, and the display that displays the measurement values at the measurement section In addition, a radio wave, for example, Bluetooth (registered trademark) can be mounted as a wireless means. With such a slide structure, since the puncture drive is received while the needle-integrated biosensor is securely held, the strength of the entire measuring apparatus can be increased. The measurement device can further include a mechanism that can recognize the left-right asymmetric structure with the puncture needle of the needle-integrated biosensor as the center line by the protruding portion of the measurement terminal.

  The puncturing drive of the measuring device is preferably a mechanism that returns quickly after tapping the upper part of the needle-integrated biosensor in the vertical direction, and preferably has a mechanism that can adjust the depth of puncturing the skin of the subject.

  The measuring device must have voice guidance and voice recognition functions for visual impairment caused by diabetes, measurement data management functions using the built-in radio clock, communication functions for medical data such as measurement data, and charging functions. Can do.

  A measurement method in the measurement unit of the measurement apparatus is not particularly limited, and potential step chronoamperometry, coulometry, cyclic voltammetry, or the like can be used.

  As described above, the needle-integrated biosensor of the present invention does not limit the user, that is, can handle a universal project.

  The needle-integrated biosensor according to the embodiment of the present invention will be described in detail with reference to the drawings. However, the present invention is not limited to the following examples unless it exceeds the gist.

FIG. 1 shows an assembly example of the biosensor of the present invention. FIG. 1a) shows a sensor with a cover, and b) shows a sensor in which a cover is not provided separately and a soft sheet also serves as a cover in the sensor a). a) i) shows a substrate 1 having a conductor 7 formed on the surface thereof, a cover 15 and a soft sheet 6 covered thereon. Here, the soft sheet 6 functions as a resist that partially covers the surface of the conductor other than the portions constituting the electrodes and terminals , and also functions as the spacer 2. FIG. 2 (ii) shows the substrate 1 having a soft sheet formed on the surface, the cover 15, and the adhesive layer 5 to be covered in the next assembly process. The soft sheet is provided so as to cover the substrate portion on the left side of the electrode reaction portion of the substrate 1 and the entire cover 15, and is also formed on the substrate excluding the electrode reaction portion and the right end portions of the conductors 7 and 7. The The adhesive layer 5 is provided on the soft sheet forming substrate excluding the electrode reaction portion and the right end portions of the conductors 7 and 7. The adhesive layer 5 also functions as the spacer 2 like the soft sheet . In iii), the cover member 15 is folded along the broken line located in the soft sheet portion connecting the substrate and the cover to the substrate 1 having the adhesive layer 5 formed on the surface, and shows a state before overlapping. In iv), a biosensor 3 that is a foldable molded body 11 formed by folding the substrate 1 and the cover member 15 is shown. Here, the cover 15 is narrower than the substrate 1, and the right end portion of the substrate 1 where the cover 15 is not formed forms terminals 11 and 11 connected to the measuring apparatus.

In this case, the flexible sheet on the substrate and the cover forms these connecting portions, and the substrate 1 and the cover 15 are folded over at the boundary, so that the folded portion is like a folded biosensor provided with a perforation on the conventional substrate. The biosensor 3 can be constructed without receiving any stress. That is, the stress applied at the time of folding is minimized by the flexibility and thinness of the soft sheet . As a result, there is an excellent feature that it is not necessary to attach a fixing tool to the folded portion or to perform a treatment such as removing the stress due to warping by thermocompression bonding.

i) to iv) of b) show an embodiment in which the cover 15 is not provided in i) to iv) of a). By taking such an embodiment, it was required in a) iii) A plate material used for the cover is not required, and an economical and low environmental load sensor is provided. Also, in b) iii), unlike a) iii), a mode in which a perforated line is provided in the soft sheet is shown. By providing a perforation on the soft sheet , further stress reduction during folding can be achieved.

FIG. 2 shows a state of the biosensor shown in FIG. 1b) in the a) AA ′ section and b) BB ′ section. Here, the electrode 10 is provided on the substrate 1, and the adhesive layer 5 and the soft sheet 6 disposed so as to sandwich the adhesive layer 5 include the sample transport path 8 and the electrode reaction part (reagent layer). It is shown that the flexible sheet is connected from the substrate 1 to the cover 6 while acting as a spacer 2 for forming the.

FIG. 3 is a view showing another assembly example of the biosensor of the present invention. a) to c) are production examples of the biosensor, i) is a constituent material required for production of the biosensor, and ii) is a molded body thereof. a) i) is a state in which the substrate 1 and the cover 15 of the biosensor are arranged in parallel to the long axis direction, and the flexible sheets 6 and 6 that serve to connect the substrate 1 and the cover 15 and also serve as the spacer 2. It is shown. Here, a conductor 7 is formed on the substrate 1. In a) ii), a molded body of each member shown in i) is shown. Here, the portion of the substrate not covered with the soft sheet 6 becomes the electrode 10, and the lower end portion of the substrate 1 on which the cover 15 is not formed forms the terminals 11 and 11 connected to the measuring device. b) i) shows the molded body shown in a) ii) and the adhesive layer disposed in the next step. The adhesive layer 5 has the function of the spacer 2 like the flexible sheet 6. b) ii) shows a state in which the adhesive layer 5 is formed on the surface of the molded body. From this state, the biosensor 3 which is the folded molded body 11 shown in c) is manufactured by folding the cover member on the substrate member with the adhesive layer 5 inside. In this case as well, the flexible sheet connects the substrate 1 and the cover 15 in the same manner as in FIG. 1, and the substrate 1 and the cover 15 are folded over at the boundary, thereby folding the biosensor having a perforation on the conventional substrate. As described above, the biosensor 3 can be constructed without being subjected to stress at the folded portion.

Fig. 4a) shows a cross-sectional view of the folded molded body 11 taken along the line AA 'shown in Fig. 3c). Here, the electrode 10 is provided on the substrate 1, and the adhesive layer 5 and the soft sheet 6 arranged so as to sandwich the adhesive layer 5 are connected to the sample transport path 8 and the electrode reaction part (reagent layer). It turns out that it is acting as the spacer 2 for forming. b) shows a BB ′ cross-sectional view of the folded compact 14 shown in FIG. 3c). Here, it is shown that the soft sheet connects the substrate 1 and the cover 6.

FIG. 5 is a view showing still another assembly example of the biosensor of the present invention. a) shows a sensor with a cover, and b) shows a sensor in which a cover is not provided separately and a soft sheet also serves as a cover in the sensor of a). The difference from FIG. 3 is the shape of the soft sheet 6, and one soft sheet corresponds to a pattern for defining the area of the electrode formed on the surface of the substrate 1 and the sample transport path 8 and the electrode reaction layer 13. The feature is that the part is cut out. By adopting such a shape, unlike the case of the soft sheet of FIG. 3, there is an effect that it is not necessary to adjust the distance between the two soft sheets each time the sensor is created.

  FIG. 6 is a diagram showing an example of use of the biosensor shown in FIG. Examples of a) and b) show usage examples of the biosensor shown in FIGS. 5 a) and b), respectively. i) shows a state before the sample solution 17 is introduced into the sample introduction port 9 of the biosensor 3, ii) shows a state during introduction, and iii) shows a state after introduction. Here, as shown in iii), the sample solution 17 does not wrap around in both the biosensors 3 a) and b). That is, as shown in FIG. 17 as an example of a conventional sensor, the problem that the sample liquid 17 spreads on the side surface of the sensor 3 near the sample introduction port 9 is solved. This is because the sample inlet 9 in the sensor 3 protrudes in a convex shape, and this eliminates the problem that the sample volume is easily influenced by the state of the sample liquid in the conventional sensor. .

  Such a state is more clearly shown by the cross-sectional view shown in FIG. a) and b) are AA ′ cross-sections (parts deviated from the electrode reaction part 13) and BB ′ cross-sections (parts of the electrode reaction part 13) when the biosensor shown in FIGS. 5a) and b) is used. ) State. i) shows the state before introducing the sample solution 17, and ii) shows the state after introducing the sample solution 17, respectively. As is apparent from ii), in the case of such a biosensor, since the vicinity of the sample introduction port 9 is convex, the sample liquid is prevented from wrapping around the side facing the sample introduction port 9, and the conventional sensor As an example, as shown in FIG. 18, there is no problem that the sample liquid wraps around the lower side of the sensor 3, that is, the side surface facing the sample introduction port 9, in addition to the sample transport path 8.

FIG. 8 is a view showing an assembly example of the suction needle integrated biosensor of the present invention. a) to d) are examples of manufacturing a needle-integrated biosensor, i) is a constituent material required for manufacturing the needle-integrated biosensor, and ii) is a molded body thereof. a) i) shows a state in which a biosensor substrate 1, a soft sheet peeling plate 31, a cover 15 are arranged, and a soft sheet 6 covered on these surfaces. Conductors 7 are disposed on the surface of the substrate 1 so as to be orthogonal to the long axis direction of the substrate 1, and the soft sheet 6 is provided with a through hole 37 for defining an electrode area. In ii), the state in which the soft sheet 6 is formed on the surface of the three members is shown from the front surface in ii) 1) and from the back surface in ii) 2). b) shows how the adhesive layer 5 is formed on the surface of the flexible sheet 6. Here, the adhesive layer 5 acts as the spacer 2 and its thickness is used to form a space for the sample transport path 8 and the vacuum space 26. Further, here, a ventilation filter 25 is provided between the sample transport path 8 and the vacuum space 26 for preventing the sample liquid from flowing into the vacuum space 26 provided for blood collection introduction. The puncture needle portion 33 is also disposed on the surface of the adhesive layer 5 provided on the cover 15. c) shows a state in which the adhesive layer 5 further covers the surface of the puncture needle portion 33 provided on the cover 15. Through this step, the puncture needle 19 can be prevented from contacting the reagent provided in the electrode reaction unit 13. The state at this time is shown from the back side in d). Here, a blood collection introduction guide 36 made of a soft material is attached to the soft sheet 6 peeled off from the peeling plate 31. Shown below these drawings is a needle-integrated biosensor 29 of the folded molded body 14 produced by a series of steps. Here, the internal pressure of the needle-integrated biosensor 29 can be evacuated by evacuating the atmosphere during the folding process. As shown in this figure, the needle-integrated biosensor 29 has an asymmetrical shape as a whole because the terminal 11 is disposed near the puncture blood collection port 32. Such a shape is characterized in that the orientation of the needle-integrated biosensor 29 is not mistaken when the needle-integrated biosensor 29 is attached to the measuring apparatus, and the terminal portion 11 is disposed near the puncture blood collection port 32. When attached to the measuring apparatus, at least the terminal portion can hold the sensor 29 reliably in addition to the lower portion of the sensor 29.

FIG. 9 shows an assembly example of the sensor of FIG. 8 in which a cover is not provided and a soft sheet also serves as a cover. 8 differs from FIG. 8 in that the cover 15 is not used separately, and the soft sheet also serves as the cover, so that the release plate 31 is used at the time of creation. 8 and 9 is a cross-sectional view along the line AA ′ (on the center line of the sensor substrate) and a cross-sectional view along the line BB ′ (on the line slightly to the right of the center of the sensor substrate). Each is shown in b).

In FIG. 10 , which is a cross-sectional view along the line AA ′, the soft sheet 6 and the electrode 10 are shown on the substrate 1. Accordingly, the periphery of the electrode 10 becomes an electrode reaction portion 13 containing a reagent and the sample transport path 8. These spaces 26 are kept in a vacuum state together with a space 26 (a space sandwiched between the soft sheet 6 and the adhesive layer 5) separated from the electrode reaction part 13 by the ventilation filter 25. For the puncture needle portion 33, the support 19 is fixed between the two adhesive layers 5, and only the tip of the needle 20 protrudes from the adhesive layer 5. In this structure, the base portion of the needle 20 is protected by the adhesive layer, thereby preventing direct contact with the reagent provided in the electrode reaction unit 13.

At the tip of the needle 20, the soft sheet 6 joins the substrate 1 and the cover 15 and folds it. As a result, the soft sheet 6 has a curved surface due to its flexibility, and a blood collection introduction guide 36 is provided on the surface. This guide 36 is provided in order to improve the close contact of the subject with the skin and to efficiently suck the blood sample after puncture using the negative pressure inside. In the BB ′ cross-sectional view, a cross section deviated from the longitudinal center of the substrate is shown, and there is no electrode reaction section 13 or sample transport path 8 and a state where the adhesive layer is filled is shown. On the other hand, the space 26 sandwiched between the lower flexible sheet 6 and the adhesive layer 5 is provided in order to secure more vacuum space.

FIG. 11 shows an example of use of the suction needle integrated biosensor 29 shown in FIG. FIG. A) shows a state before the skin 27 of the subject is punctured. Here, the blood collection guide 36 provided at the puncture blood collection port 32 of the biosensor 29 integrated with the skin 27 is in close contact with the skin 27. b) shows a state where the tip of the needle-integrated biosensor 29 is pressed against the skin 27 and punctured. Here, the soft sheet 6 is recessed due to its flexibility, so that the puncture needle 20 fixed inside the needle-integrated biosensor 29 breaks through the soft sheet 6. In c), after the puncture, the puncture needle 20 returns to the original position by the restoring force of the soft sheet 6, and the blood collection 24 is sucked by the negative pressure inside the sensor 3 through the penetration (blood collection) port 32 formed at that time. The state where the sample is promptly introduced into the sample conveyance path and the electrode reaction part is shown. Further, the blood collection 24 is stopped from flowing into the sensor 3 at the ventilation filter 25. In (d), the state where the puncture / blood collection has been completed and the component measurement is in progress is shown. Here, a state in which the bleeding 24 from the skin 27 is minimized by the attachment of the blood collection introduction guide is shown.

FIG. 12 shows an example of manufacturing a suction-type needle integrated biosensor 29. The feature of the aspirating needle-integrated biosensor 29 is that by providing the terminal 11 portion below, the substrate 1 has a simple rectangular shape, and more sensors can be manufactured from a single plate. a) to g) show the respective manufacturing steps. In a), the biosensor substrate 1, the soft sheet release plate 31, and the cover 15 are arranged close to each other. The release plate 31 is provided with a mold for taking the molding material of the soft sheet 6 into a mold and forming a blood collection introduction guide. On the surface of the substrate 1 conductors 7,7 for forming the electrodes are six pairs formed, soft sheet 6 is shown to be coated to the surface of the peeling plate 31 and the cover 15 of the flexible sheet. The soft sheet is provided with a through hole 37 for defining the electrode area. b) shows a state in which the soft sheet 6 is formed on the surfaces of the three members when viewed from the front surface (b) i)) and when viewed from the back surface (b) ii)). In the next step shown in c), a cutting line is provided for each biosensor unit 39 so that the substrate 1 and the cover 15 member can be easily separated, and then bonded to the surface of the soft sheet 6 in the step d). After the agent layer 5 is formed, the soft sheet 6 is peeled off from the peeling plate 31 on the back side. As shown in d) ii), a blood collection introduction guide 36 provided on the peeling plate 31 is formed on the back surface of the portion sandwiched between the substrate 1 and the cover 15. e) i) shows how the puncture needle 33 is fixed inside the sensor (cover 15 portion). In this state, by folding the substrate 1 and the cover 15 with the connecting portion between the substrate and the cover of the soft sheet 6 as a boundary, six needle integrated biosensor units 40 as shown in f) are formed. Are separated into individual needle integrated biosensors 29 as shown in g). Here, the internal pressure of the needle-integrated biosensor unit 40 can be evacuated by evacuating the atmosphere during the folding step f).

The needle-integrated biosensor shown in FIG. 13 is different from the needle-integrated biosensor shown in FIG. 12 and does not include a suction mechanism and uses only a capillary phenomenon for blood collection. Constituently, as shown in c) i) and ii), a cutting line is provided on the substrate 1 and the cover 15 member for each biosensor unit 39, and at the same time, the substrate 1 is close to the electrode 10 A punched through hole 37 is provided. The through hole 37 is for discharging air, and blood can be collected by capillary action. Further, the electrode area can be defined by the through hole 37. Further, in the next step f), a mode is shown in which the flexible sheet 6 is arranged to fix the support 19 portion of the puncture needle portion 33 and to separate the base portion of the needle 20 from the reagent of the electrode reaction portion 13. ing.

  FIG. 14 shows a front view a) i), a rear view a) ii) and a centerline cross-sectional view b) of the individual needle-integrated biosensor 29 manufactured in FIG. In a) ii) and b), a through hole 37 penetrating to the inside of the biosensor is shown.

FIG. 15 is different from the needle-integrated biosensor shown in FIG. 12 in that a cover 15 is not provided separately and a soft sheet also serves as a cover. Even if the cover 15 is not provided separately, many sensors can be manufactured from a single plate.

  The biosensor according to the present invention is a home-use self-diagnosis blood glucose meter, urine sugar meter, glycated hemoglobin meter, lactic acid meter, cholesterol meter that electrochemically measures the component concentration of various liquids using an enzyme or the like. It is effectively used as a biosensor for use in uric acid meters, protein meters, single nucleotide polymorphism sensors, DNA chips used for genetic diagnosis, as well as alcohol meters, glutamic acid meters, pyruvic acid meters, pH meters and the like.

It is a figure which shows one assembly example of the biosensor which concerns on this invention. It is a figure which shows an example of the cross section of the biosensor which concerns on this invention. It is a figure which shows the other assembly example of the biosensor which concerns on this invention. It is a figure which shows the other example of the cross section of the biosensor which concerns on this invention. It is a figure which shows the further another assembly example of the biosensor which concerns on this invention. It is a figure which shows one usage example of the biosensor which concerns on this invention. It is a figure which shows the usage example of the biosensor which concerns on this invention in a cross section. It is a figure which shows one assembly example of the suction blood collection type | mold needle integrated biosensor which concerns on this invention. It is a figure which shows the other assembly example of the suction blood collection type | mold needle integrated biosensor which concerns on this invention. It is a figure which shows an example of the cross section of the suction blood collection type | mold needle integrated biosensor which concerns on this invention. It is a figure which shows the usage example of the suction blood collection type | mold needle | hook integrated biosensor which concerns on this invention in a cross section. It is a figure which shows the other manufacture example of the needle | hook integrated biosensor which concerns on the present invention at the blood collection type. It is a figure which shows the other manufacture example of the needle | hook integrated biosensor which concerns on this invention. It is a figure which shows an example of the cross section of the needle | hook integrated biosensor which concerns on this invention. It is a figure which shows one manufacture example of the needle-integrated biosensor according to the present invention. It is a figure which shows one assembly example of the conventional biosensor. It is a figure which shows one usage example of the conventional biosensor. It is a figure which shows the usage example of the conventional biosensor in a cross section.

DESCRIPTION OF SYMBOLS 1 Board | substrate 2 Spacer 3 Biosensor 4 Connection part 5 Adhesive layer 6 Soft sheet 7 Conductor 8 Sample conveyance path 9 Sample inlet 10 Electrode 11 Terminal 12 Air outlet 13 Electrode reaction part (reagent layer)
14 Folding Molded Body 15 Cover 16 Perforation 17 Sample Solution 19 Puncture Needle Support 20 Puncture Needle 24 Blood Collection 25 Ventilation Filter 26 Vacuum Space 27 Skin 28 Open Space 29 Needle-Integrated Biosensor 31 Peeling Plate 32 Puncture Blood Collection Port 33 Puncture Needle Portion 36 Blood collection introduction guide 37 Through hole 38 Blood collection introduction guide forming template 39 Biosensor unit 40 Needle-integrated biosensor unit

Claims (20)

In a biosensor in which electrodes, spacers, and a cover are sequentially formed on an insulating substrate,
Insulating substrate and the cover are connected by a flexible sheet, form a part of the resist layer of the spacer with some soft matter sheet acts as a resist layer covering the conductor surface other than portions constituting the electrodes and terminals A biosensor characterized in that it is folded at a soft sheet portion connecting the insulating substrate and the cover, and the electrode is housed inside the biosensor. The cover is made of the same material as the soft sheet forming material, and the cover and the soft sheet are integrated, so that the soft sheet connects the insulating substrate and the cover. Biosensor. An insulating substrate and a cover on the soft sheet portion that Ide joint, according to claim 1 or 2, wherein the biosensor sample inlet for introducing a sample to be measured is provided. The biosensor according to claim 1 or 2, wherein the soft sheet material is made of vinyl / acrylic resin, urethane acrylate resin, polyester acrylate resin, polyvinyl chloride, polyethylene, polyester, polyolefin, polyvinyl fluoride, or cellophane.   The biosensor according to claim 4, wherein the acrylic resin is a thermosetting type or a photocurable type.   The biosensor according to claim 1 or 2, wherein the resist layer has a thickness of 5 to 500 µm.   The biosensor according to claim 1 or 2, wherein the resist layer contains at least one reagent selected from an enzyme, a mediator, and a surfactant.   The biosensor according to claim 1 or 2, wherein at least one reagent layer of an enzyme, a mediator or a surfactant is formed on the electrode.   The biosensor according to claim 1 or 2, wherein at least one of the substrate and the cover is an oxygen permeable material.   A needle-integrated biosensor in which a puncture needle is further fixed between the resist layer and the cover of the biosensor according to claim 1 or 2.   The biosensor according to claim 10, wherein the puncture needle is fixed with an adhesive. At the time of puncturing the subject, the puncture needle breaks through the soft sheet portion connecting the insulating substrate and the cover, pierces the subject's skin, and then the puncture needle is pulled back from the subject's skin by the restoring force of the soft sheet material, The biosensor according to claim 11, wherein the body fluid is introduced into the sample transport path of the biosensor through the through hole of the soft sheet portion generated at that time.   The biosensor according to claim 10 or 11, wherein the puncture needle is isolated from a reagent layer provided on the electrode by a resist layer or an adhesive layer. The needle-integrated biosensor according to claim 10, wherein a blood collection inlet guide is provided on a soft sheet connecting the insulating substrate and the cover.   The biosensor according to claim 10, wherein the biosensor is maintained at a negative pressure or a vacuum, and blood is collected using the suction force during puncture blood collection. The biosensor according to claim 1, wherein after the soft sheet layer and the adhesive layer are sequentially formed on the surface of the insulating substrate and the cover on which the electrode is formed, the insulating substrate and the cover are arranged so that the electrode is placed inside the biosensor. The biosensor is manufactured by folding the insulating substrate and the cover at the soft sheet portion connecting the two . After disposing a release plate between the insulating substrate on which the electrode is formed and the cover, a soft sheet layer is formed by a screen printing method, a thermocompression laminating method, or a method using an adhesive tape. The biosensor manufacturing method according to claim 16, wherein the release plate is peeled off when the soft sheet portions to be joined are folded. 3. The biosensor according to claim 2, wherein a soft sheet layer and an adhesive layer integrally formed with the cover portion are sequentially formed on the surface of the insulating substrate on which the electrode is formed, and then the electrode is insulated so that the electrode is accommodated in the biosensor. Of a biosensor manufactured by folding an insulating substrate and a cover at a soft sheet portion connecting the conductive substrate and the cover. After the release plate is placed on the insulating substrate on which the electrode is formed and the cover is to be formed adjacent to the insulating substrate, the soft sheet layer that is integrated with the cover part uses screen printing, thermocompression laminating, or adhesive tape. 19. The method for producing a biosensor according to claim 18, wherein the release plate is peeled off when the flexible sheet portion formed by the method is further folded and the soft sheet portion connecting the insulating substrate and the cover portion is folded.   20. The method for producing a needle-integrated biosensor according to claim 16, wherein a puncture needle is fixedly disposed on the adhesive layer.

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