Patent Application
20100300898
The present invention relates to a technology for detecting whether or not a sample supplied to an analysis tool is insufficient when the sample is analyzed using the analysis tool.
When the glucose concentration within blood is measured, a method of using a disposable analysis tool is being employed as a simple and easy technique. The analysis tool includes, for example, an electrode-type biosensor 9 shown in
The electrode 91 has an working electrode 94 for performing transfer of electrons to/from a certain component within blood and a counter electrode 95 for generating an electric potential difference between the electrode 92 and the working electrode 94. The working electrode 94 and the counter electrode 95 are exposed in the flow channel 93.
In such a biosensor 9, when a voltage is applied between the working electrode 94 and the counter electrode 95, a response electric current is output in response to a concentration of a certain component within the sample. Therefore, in the biosensor 9, it is possible to measure the glucose concentration or the like by measuring the response electric current using the working electrode 94 (electrode 91) and the counter electrode 95 (electrode 92).
The biosensor 9 is provided with a detection electrode 96 for detecting whether or not the sample is appropriately supplied in addition to the electrodes 91 and 92 for analyzing blood. In such a biosensor 9, it is determined that a sufficient amount of the sample such as blood has been supplied to the flow channel 93 when it is identified that a liquid junction is formed between the detection electrode 96 and the first electrode 92 or the second electrode 93.
However, as shown in
The present invention has been made to appropriately detect whether or not the sample supplied to the analysis tool is short when the sample is analyzed using the analysis tool.
According to a first aspect of the present invention, there is provided an analysis tool including: a substrate; and plural electrodes formed on the substrate, wherein the plural electrodes include two or more first electrodes having an working electrode and one or more second electrodes having a counter electrode.
The analysis tool according to the present invention may further include a flow channel for transferring the sample. In this case, it is preferable that the plural electrodes are arranged such that the working electrode and the counter electrode have a symmetrical positional relationship in the movement direction of the sample in the flow channel.
The number of the one or more second electrodes is equal to the number of the two or more first electrodes. When the number of each of first and second electrodes is 2, plural electrodes may be arranged side by side in the order of the counter electrode, the working electrode, the working electrode, and the counter electrode in the movement direction, or in the order of the working electrode, the counter electrode, the counter electrode, and the working electrode.
In addition, the plural electrodes may include two first electrodes and one second electrode. In this case, for example, the electrodes are arranged side by side in the order of the working electrode, the counter electrode, and the working electrode with respect to the movement direction.
According to a second aspect of the present invention, there is provided an analyzer for performing analysis of a sample using an analysis tool, the analysis tool having first and second working electrodes and one or more counter electrodes, the analyzer including: an electric current measurement means for detecting a response electric current when a voltage is applied between first and second working electrodes and the counter electrode; a detection means for detecting shortage of the sample supplied to the analysis tool by comparing a first response electric current obtained when a voltage is applied between the first working electrode and the counter electrode and a second response electric current obtained when a voltage is applied between the second working electrode and the counter electrode; and an analysis means for performing analysis of the sample based on at least one of the first and second response electric currents.
According to a third aspect of the present invention, there is provided a method of detecting whether or not the sample supplied to an analysis tool is insufficient when the sample is analyzed using the analysis tool, the analysis tool having first and second working electrodes and one or more counter electrode, wherein it is determined whether or not the amount of sample supplied to the analysis tool is insufficient by comparing a first response electric current obtained when a voltage is applied between the first working electrode and the one or more counter electrodes and a second response electric current obtained when a voltage is applied between the second working electrode and the one or more counter electrode.
In the detection method according to the present invention, for example, it is determined that the sample supplied to the analysis tool is insufficient when the difference between the first and second response electric currents is not within a predetermined range.
For example, the one or more counter electrodes include first and second counter electrodes. In this case, the first response electric current is measured when a voltage is applied between the first working electrode and the first counter electrode, and the second response electric current is measured when a voltage is applied between the second working electrode and the second counter electrode.
For example, the analysis tool has a flow channel for moving the sample, and the first and second working electrodes and the one or more counter electrodes are arranged symmetrically in a movement direction of the sample.
According to a fourth aspect of the present invention, there is provided a method of supplying a sample and analyzing a sample using an analysis tool, the analysis tool having first and second working electrodes and one or more counter electrodes, wherein shortage of the sample supplied to the analysis tool is detected by comparing a first response electric current obtained when a voltage is applied between the first working electrode and the one or more counter electrodes and a second response electric current obtained when a voltage is applied between the second working electrode and the one or more counter electrodes, and the sample is analyzed based on at least one of the first and second response electric currents.
Hereinafter, the present invention is described below in detail with reference to the accompanying drawings.
First, a first embodiment of the present invention is described with reference to
The biosensor 1 shown in
The substrate 10 is formed in a shape larger than the cover 12 using an insulation resin material such as PET, and has electrodes 14, 15, 16, and 17 and a reagent layer 18 formed on the surface thereof.
The electrodes 14 to 17 are formed to have a band shape extending in a longitudinal direction D3 and D4 of the substrate 10 and having substantially the same width. These electrodes 14 to 17 have terminal sections 14A, 15A, 16A, and 17A exposed in a lateral direction of the cover 12. The terminal sections 14A to 17A are to make contact with the first to fourth terminals 20A to 20D of the analyzer 2 when the biosensor 1 is installed in the analyzer 2 as described below with reference to
The electrodes 14 and 15 further have working electrodes 14W and 15W for performing transfer of electrons to/from a certain component within the sample. Meanwhile, the electrodes 16 and 17 have counter electrodes 16C and 17C for generating an electric potential difference from the working electrodes 14W and 15W. The working electrodes 14W and 15W and the counter electrodes 16C and 17C are arranged side by side in the order of the counter electrode 17C, the working electrode 15W, the working electrode 14W, and the counter electrode 16C in the direction D1 inside the capillary 13.
These electrodes 14 to 17 may be formed by CVD, sputtering, or deposition using gold, platinum, palladium, nickel, or the like, or by forming a conductive film through a screen printing using carbon inks and then irradiating laser light to provide a slit.
The reagent layer 18 is provided to cover the working electrodes 14W and 15W and the counter electrodes 16C and 17C in series inside the capillary 13. This reagent layer 18 includes, for example, an oxidoreductase and an electron carrier material, and is formed in a solid state that can be readily dissolved in a liquid sample such as blood.
The oxidoreductase is selected depending on the type of the analysis target component within the sample. For example, when glucose is analyzed, glucose oxidase (GOD) or glucose dehydrogenase (GDH) may be used, and typically, PQQGDH is used. The electron carrier material may include, for example, a ruthenium complex or an iron complex, and typically [Ru(NH3)6]Cl3 or K3[Fe(CN)6].
A pair of spacers 11 are provided to define a distance from the surface of the substrate 10 to the lower surface of the cover 12, i.e., the height of the capillary 13, and are configured of, for example, a double-face adhesive tape or a hot-melt film. The spacers 11 extend in a width direction D1 and D2 of the substrate 10 and are also arranged to be separated in a longitudinal direction D3 and D4 of the substrate 10. In other words, the pair of spacers 11 define the width of the capillary 13 and the area (the contact area making contact with the sample) of the portion exposed within the capillary 13 (the working electrodes 14W and 15W and the counter electrodes 16C and 17C) in the electrodes 14 to 17. In addition, since the electrodes 14 to 17 have substantially the same width, the areas of the working electrodes 14W and 15W and the counter electrodes 16C and 17C are set to be substantially the same. Here, the areas are referred as being substantially the same considering the irregularity of the area caused by a manufacturing error or the like.
The cover 12 is to define the capillary 13 in association with the spacers 11 or the like. This cover 12 is formed of the same material as that of the substrate 10 such as PET or thermoplastic resin having a high wettability such as vinylon or high-crystalline PVA.
The capillary 13 is to move the introduced sample such as blood in a width direction D1 and D2 of the substrate 10 using a capillary action and maintain the introduced sample. In other words, in the capillary 13, when the sample is introduced, the sample moves while discharging gas within the capillary 13. In this case, inside the capillary 13, the reagent layer 18 is dissolved so as to provide a liquid-phase reaction system including certain elements such as an oxidoreductase, an electron carrier material, and glucose.
The analyzer 2 shown in
First to fourth terminals 20A to 20D are to make contact with the terminal sections 14A to 17A of the electrodes 14 to 17 of the biosensor 1 when the biosensor 1 is installed in the analyzer 2.
The voltage application unit 21 is to apply a voltage between the terminal sections 14A to 17A of the biosensor 1 through the first to fourth terminals 20A to 20D. This voltage application unit 21 is configured of, for example, a direct current power source.
The electric current measurement unit 22A is to measure a first response electric current when a voltage is applied between the terminals 14A and 16A (between the working electrode 14W and the counter electrode 16C) of the biosensor 1 by the voltage application unit 21. The electric current measurement unit 22B is to measure a second response electric current when a voltage is applied between the terminal sections 15A and 17A (between the working electrode 15W and the counter electrode 17C) of the biosensor 1 from the voltage application unit 21.
The detection unit 23 is to detect whether or not the sample is supplied to the capillary 13 of the biosensor 1 based on the first and second electric currents measured by the electric current measurement unit 22A and 22B after the biosensor 1 is installed in the analyzer 2.
The control unit 24 is to control the voltage application unit 21 and a voltage state applied between the working electrodes 14W and 15W and the counter electrodes 16C and 17C.
The computation unit 25 computes a concentration of a certain component within the reagent solution or a correction value required in this computation in response to at least one of the first and second response electric current values measured by the electric current measurement units 22A and 22B. The computation unit 25 has, for example, a timer function and stores a concentration of a certain component and a response electric current (or an equivalent value (for example, a voltage value) corresponding to the response electric current) to compute the concentration of a certain component based on the response electric current after a predetermined time, for example, from starting applying the voltage.
While each of the detection unit 23, the control unit 24, and the computation unit 25 includes, for example, a CPU and memory (such as ROM or RAM), all of the detection unit 23, the control unit 24, and the computation unit 25 may be configured by connecting plural memory devices to a single CPU.
Next, a method of analyzing the sample using the biosensor 1 and the analyzer 2 is described below.
In the sample analysis, first, the biosensor 1 is installed in the analyzer 2, and the sample such as blood is introduced from the end portion of the capillary 13 of the biosensor 1 into the inner side of the capillary 13. Since both ends of the capillary 13 are opened at the outer side of the analyzer 2 when the biosensor 1 is installed in the analyzer 2, the sample can be introduced from either end. The sample introduced into the capillary 13 is moved into the direction D1 or D2 within the capillary 13.
Meanwhile, the analyzer 2 applies a voltage between the electrodes 14 to 17 by controlling the voltage application unit 21 using the control unit 24 and measures the response electric current at that moment using the electric current measurement units 22A and 22B. The voltage value applied by the voltage application unit 21 is set to, for example, a constant voltage of about 200 mV. The voltage is continuously applied to the electrodes 14 to 17 before the sample is introduced into the capillary 13. Applying the voltage may be stopped only for a predetermined time (for example, 0.5 to 60 seconds) after a liquid junction between the working electrodes 14W and 15W and the counter electrodes 16C and 17C is identified, and then the voltage may again be applied to the electrodes 14 to 17.
The detection unit 23 compares first and second response electric currents measured by the electric current measurement units 22A and 22B after a predetermined time (for example, 5 seconds) is elapsed from starting applying the voltage between the electrodes 14 to 17. In this case, it is determined whether or not the difference between the first and second response electric currents is within a predetermined range. Here, the predetermined range is determined arbitrarily, but may be appropriately set depending on the size of the capillary 13, the areas of the working electrodes 14W and 15W, a composition of the reagent layer 18, the type of the analysis component, the magnitude of the applied voltage, or the like.
When it is determined by the detection unit 23 that the difference between the first and second response electric currents is within a predetermined range, the detection unit 23 determines that an amount of the sample sufficient to analyze the sample is supplied to the capillary 13. When the difference between the first and second response electric currents is within a predetermined range, it means that a liquid junction state between the working electrode 14W and the counter electrode 16C is approximately the same as a liquid junction state between the working electrode 15W and the counter electrode 17C as shown in
Meanwhile, in the detection unit 23, when it is determined that the difference between the first and second response electric currents is not within a predetermined range, the detection unit 23 determines that the amount of the sample S sufficient to analyzes the sample S is not supplied to the capillary 13. When the difference between the first and second response electric currents is not within a predetermined range, a liquid junction state between the working electrode 14W and the counter electrode 16C is different from a liquid junction state between the working electrode 15W and the counter electrode 17C as shown in
In the detection unit 23, when it is determined that the difference between the first and second response electric currents is not within a predetermined range, it is determined that an amount of the sample sufficient to perform the analysis is not supplied to the capillary 13 (the sample is insufficient), and an error process for the sample shortage is performed. The detection unit 23 re-compares the first and second response electric currents and re-detects whether or not the sample is insufficient after it is determined that the difference between the first and second response electric currents is not within a predetermined range after a predetermined time has elapsed.
On the contrary, in the detection unit 23, when it is determined that the difference between the first and second response electric currents is within a predetermined range, it can be determined that the amount of the sample sufficient to perform the analysis is supplied to the capillary 13. Therefore, the analysis of a certain component within the sample is performed.
The analysis of a certain component within the sample is performed based on the first and second response electric currents when a voltage is applied by the voltage application unit 21 between the electrodes 14 to 17. More specifically, first, the computation unit 25 samples the first and second response electric currents measured by the electric current measurement units 22A and 22B after the sample is detected, or the voltage is re-applied, and then, a predetermined time is elapsed. Next, the computation unit 25 computes the concentration of a certain component by applying at least one of the first and second response electric currents to a calibration curve or a reference table representing a relationship between the concentration of a certain component and the response electric current value.
Here, when the analysis is based on both the first and second response electric currents, for example, an average value or an integration value of the first and second response electric currents is employed.
As described above, in the analysis of the sample using the biosensor 1 and the analyzer 2, the shortage of the sample is detected based on two working electrodes 14W and 15W lined in the movement direction D1 and D2 of the sample. Therefore, as long as each of the working electrodes 14W and 15W does not appropriately make contact with the sample, it is determined that the sample supplied to the capillary 13 is insufficient. Therefore, when it is determined that the sample sufficient to perform the analysis is supplied, each of the working electrodes 14W and 15W makes contact with the sample over substantially the entire area thereof. As a result, it is possible to prevent the analysis of the sample from being performed without considering the shortage of the sample supplied to the capillary 13, and improve the measurement accuracy.
In addition, since the working electrodes 14W and 15W and the counter electrodes 16C and 17C are arranged side by side symmetrically in the movement direction D1 and D2 of the sample, the shortage of the sample is detected in the same condition even when the sample is supplied to either end of the capillary 13. Therefore, it is possible to appropriately detect the supply shortage of the sample irrespective of the introduction direction of the sample into the capillary 13.
The present invention is not limited to the aforementioned embodiments, but may be variously changed. For example, the arrangement or the number of the working electrodes and the counter electrodes may have the configurations shown in
In the example shown in
In the example shown in
Also in the example shown in
In addition, since the working electrodes 14W′ and 15W′ (14W″ and 15W″) and the counter electrodes 16C′ and 17C′ (16C″) are arranged side by side symmetrically in the movement direction D1 and D2 of the sample, it is possible to appropriately detect the supply shortage of the sample irrespective of the introduction direction of the sample into the capillary 13.
Next, the second embodiment of the present invention is described below with reference to
Similar to the biosensor 1 described above (refer to
Electrodes 44, 45, 46, and 47 are formed on the substrate 40. The electrodes 44 to 47 have curved portions 44A, 45A, 46A, and 47A extending in the directions of D1 and D2 and lead portions 44B, 45B, 46B, and 47B extending in directions of D3 and D4. The curved portions 44A to 47A are arranged side by side in the directions of D3 and D4, and include the working electrodes 44W and 45W and the counter electrodes 46C and 47C defined by the spacer 41.
Inside the capillary 43, the working electrodes 44W and 45W and the counter electrodes 46C and 47C are arranged side by side in the order of the counter electrode 47C, the working electrode 45W, the counter electrode 46C, and the working electrode 44W in the direction D4.
The spacer 41 is to define a distance from the upper surface of the substrate 40 to the lower surface of the cover 42, i.e., the height of the capillary 43, and has a slit 48. The slit 48 defines the width of the capillary 48 for introducing the sample and the area of the portion (including the working electrodes 44W and 45W and the counter electrodes 46C and 47C) exposed inside the capillary 43 in the electrodes 44 to 47.
Here, the capillary 43 is to move the introduced sample such as blood in the direction D4 using a capillary action and retain the introduced sample. In the inner side thereof, the reagent layer 49 is formed to cover at least the working electrodes 44W and 45W. Such a spacer 41 is configured of, for example, a double-face adhesive tape or a hot-melt film.
The cover 42 is to define the capillary 43 in association with the spacer 41 or the like, and has a through-hole 42A for discharging the gas inside the capillary 43. The cover 42 is formed of the same material as that of the substrate 40, such as thermoplastic resin or PET having a high wettability such as vinylon or high-crystalline PVA.
In the biosensor 4, similar to the biosensor 1 (refer to
Also in the biosensor 4 shown in
Next, examples of the present invention is described.
In this example, whether or not the shortage of the sample can be detected in the biosensor according to the present invention was reviewed.
(Process of Manufacturing Biosensor)
A biosensor having the same configuration as that of the biosensor 1 described with reference to
(Measurement of Response Electric Current)
The response electric current was measured using the electric current meter by applying a constant DC voltage of 200 mV between the working electrode and the counter electrode after 1 second from detecting the liquid junction by setting, to 0 second, a timing when the sample is supplied to the capillary, and then, a liquid junction is detected between the working electrode and the counter electrode. As the sample, a glucose solution having the glucose concentration of 600 mg/dL was used. As the measurement result for the response electric current, the result (the sample number N=2) obtained when a glucose solution having an amount sufficient to fill the capillary is supplied is represented in
As recognized from
Meanwhile, as recognized from
Therefore, it was verified that it is possible to detect a supply shortage of the sample within the capillary by measuring the response electric currents using two working electrodes and comparing those response electric currents.
| Number | Date | Country | Kind |
|---|---|---|---|
| 2007-282783 | Oct 2007 | JP | national |
This application is the National Phase of International Application No. PCT/JP2008/069983, filed 31 Oct. 2008, which claims priority to and the benefit of JP patent application number 2007-282783, filed 31 Oct. 2007, the contents of all which are incorporated by reference herein.
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/JP2008/069983 | 10/31/2008 | WO | 00 | 7/14/2010 |
