JP2010156639A - Method and system of determining oxidation stress state - Google Patents
Method and system of determining oxidation stress state Download PDFInfo
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- Investigating Or Analysing Biological Materials (AREA)
Abstract
Description
本発明は、例えば、8-ヒドロキシデオキシグアノシンのよう低分子物質の測定を行うのに適したイムノクロマトセンサに関する。 The present invention relates to an immunochromatographic sensor suitable for measuring a low molecular substance such as 8-hydroxydeoxyguanosine.
「酸化ストレス状態」とは、体内で活性酸素と抗酸化物質とのバランスが崩れて活性酸素が優位になっている状態をいう。
喫煙、睡眠不足等の身体への過剰の負荷、過度のストレスが原因で抗酸化作用が低下すると、活性酸素が増加して酸化ストレスが高い状態になる。
“Oxidative stress state” refers to a state in which active oxygen is dominant because the balance between active oxygen and antioxidants is lost in the body.
When the antioxidant action decreases due to excessive load on the body such as smoking and lack of sleep and excessive stress, active oxygen increases and oxidative stress becomes high.
酸化ストレスが高い状態は、上記したように、喫煙、睡眠不足等の身体への過剰の負荷、過度のストレスが原因で陥る症状であるため、これが長く続くと何らかの病気に進行していく可能性がある。また、体内の細胞の酸化が進むと身体が老化するという問題もある。
このため、酸化ストレス状態を出来るだけ早期に検知することによって、このままでは病気に進行していく可能性がある状態、即ち、未病の状態を検知することができる。
一方、酸化ストレス状態を調べる方法には、いくつか知られており、測定もされている。しかし、酸化状態を測定するために、生体試料を安定にするための前処理が必要であったり、煩雑な測定操作をしなければならないという問題点がある。
本発明は、上記した酸化ストレス状態を出来るだけ早期に検知するための酸化ストレス状態判定方法及びシステムに関する。
As described above, a state with high oxidative stress is a symptom caused by excessive stress on the body such as smoking and lack of sleep and excessive stress. There is. Another problem is that the body ages as cells in the body oxidize.
For this reason, by detecting the oxidative stress state as early as possible, it is possible to detect a state where there is a possibility of progressing to a disease, that is, a non-disease state.
On the other hand, several methods for examining the oxidative stress state are known and measured. However, in order to measure the oxidation state, there is a problem that a pretreatment for stabilizing the biological sample is necessary or a complicated measurement operation is required.
The present invention relates to an oxidative stress state determination method and system for detecting the above oxidative stress state as early as possible.
上記した目的を達成するために本発明に係る酸化ストレス状態判定方法は、生体試料サンプル中の8-ヒドロキシデオキシグアノシン(8-OHdG)の濃度を、生体試料を採取した直後の早い時間で測定し、測定した8-ヒドロキシデオキシグアノシンの濃度に基づいて、体内の酸化ストレス状態を判定することを特徴とする。
生体試料サンプルは、生体内での状態をそのまま反映できるものでなければならない。生体試料を採取後に外気に接触し、また種々の外的要因を受けて時間が経過することで状態が変化し、間違った結果を得ることにつながることを回避するために、必要に応じて、前記生体試料サンプルは、前処理をせずに測定に供することができ、また、生体試料サンプルを測定試料と混合してから測定に供することもできる。
生体試料サンプルとしては、例えば、尿や全血等が用いられる。
また、生体試料サンプル中の8-ヒドロキシデオキシグアノシン(8-OHdG)の濃度の測定では、DNAの構成物質であるグアノシンに対する酸化損傷の状態を知ることができるが、その損傷は酸化作用のある物質からの酸化作用により起こるもので、酸化作用の蓄積が8-ヒドロキシデオキシグアノシンの濃度に関係する。それとともに酸化作用の異なる前記8-ヒドロキシデオキシグアノシンとは別の分析物の濃度も測定し、両測定結果に基づいて、酸化ストレス状態を総合的に判定してもよい。
この場合、別の分析物は、例えば、クロモグラニンＡ、アミラーゼ、酸素ラジカル、バイオピリン、チオレドキシン、ＭＤＡ−ＬＤＬ、血中酸化型α１アンチトリプシン、酸化ＬＤＬ受容体ＬＯＸ−１からなる群から選択され得る。
具体的には、例えば、生体試料サンプル中に存在する8-ヒドロキシデオキシグアノシンと、該8-ヒドロキシデオキシグアノシンに特異的に反応する反応物質とを反応させ、反応により生成又は消費される物質の濃度を電気的又は光学的に測定することにより、生体試料サンプル中の8-ヒドロキシデオキシグアノシンの濃度が測定され得る。
生体試料サンプル中に存在する8-ヒドロキシデオキシグアノシンの濃度と、酸化損傷を受けることなく生体試料サンプル中に存在するデオキシグアノシンの濃度とを比較することによって酸化ストレス状態の判定を行ってもよい。また、生体試料サンプル中に既知濃度のデオキシグアノシンを添加し、デオキシグアノシンの添加前後の8-ヒドロキシデオキシグアノシンの濃度およびデオキシグアノシンの濃度を比較することによって酸化ストレス状態の判定を行ってもよい。
本発明に係る酸化ストレス状態判定システムは、生体試料サンプルを導入するサンプル導入部と、サンプル導入部から導入された生体試料サンプル中の8-ヒドロキシデオキシグアノシンの濃度を測定する8-ヒドロキシデオキシグアノシン濃度測定部と、前記濃度測定部において測定された情報を入力し、8-ヒドロキシデオキシグアノシン濃度から体内の酸化ストレス状態を判定する判定処理部とを備えていることを特徴とする。
生体試料サンプルとしては、例えば、尿が用いられ得る。
前記8-ヒドロキシデオキシグアノシン濃度測定部は、生体試料サンプルと、8-ヒドロキシデオキシグアノシンに特異的に反応する反応物質とを反応させることができるように構成され得る。この場合、前記8-ヒドロキシデオキシグアノシン濃度測定部は、予め前記反応物質を備えていてもよく、また、前記反応物質を測定部の外部から供給する反応物質供給部を備えていてもよい。
前記8-ヒドロキシデオキシグアノシンに特異的に反応する反応物質は、抗8-ヒドロキシデオキシグアノシン抗体による抗原抗体反応を生じる試薬、又はアプタマーによる結合反応を生じる試薬であり得る。
前記8-ヒドロキシデオキシグアノシン濃度測定部は、前記反応物質との反応により生成又は消費される物質の濃度を電気的又は光学的に測定することができるように構成され得る。
前記8-ヒドロキシデオキシグアノシン濃度測定部における前記光学的に測定を行う手段は、例えば、分光光度計である。この場合、前記分光光度計は、8-ヒドロキシデオキシグアノシン濃度測定部におけるサンプルの反射率、蛍光、吸光度、透過率又はエバネッセント波の少なくとも一つを測定するために用いられ得る。
前記8-ヒドロキシデオキシグアノシン濃度測定部における前記電気的に測定を行う手段は、例えば、電気化学センサである。この場合、前記電気化学センサは、8-ヒドロキシデオキシグアノシン濃度測定部におけるサンプルの電流値、電圧値、電気抵抗値又はインピーダンスの少なくとも一つを測定するために用いられ得る。
前記8-ヒドロキシデオキシグアノシン濃度測定部は、緩衝液を含有する液透過性緩衝液層と、前記反応物質を含有する反応層と、生体試料サンプルが、前記液透過性緩衝液層を通過した後に前記反応層に至るように前記液透過性緩衝液層及び前記反応層を支持する支持部材を有し得る。
前記8-ヒドロキシデオキシグアノシン濃度測定部は、前記緩衝液層と液接触する少なくとも一つの付加的な層をさらに含み得る。具体的には、前記付加的な層は、血球分離層、放射線遮断層、干渉除去層、透析層又は濾過層の少なくとも一つであり得る。
前記支持部材は、生体試料サンプル導入孔を備え、かつ、前記支持部材は、前記緩衝液層及び前記反応層を挟み込み、前記サンプル導入孔から生体試料サンプルが前記緩衝液層及び前記反応層に供給されるように構成され得る。
前記8-ヒドロキシデオキシグアノシン濃度測定部は、色素を保持する色素層を備え、前記緩衝液層は、前記色素層の上に重ねて配置されるか、又は前記色素層と並置され得る。
In order to achieve the above object, the oxidative stress state determination method according to the present invention measures the concentration of 8-hydroxydeoxyguanosine (8-OHdG) in a biological sample sample at an early time immediately after collecting the biological sample. The oxidative stress state in the body is determined based on the measured concentration of 8-hydroxydeoxyguanosine.
The biological sample sample must be able to reflect the state in the living body as it is. In order to avoid contact with the outside air after collecting a biological sample, and changing the state over time due to various external factors, leading to wrong results, as necessary, The biological sample sample can be subjected to measurement without pretreatment, and can also be subjected to measurement after mixing the biological sample sample with the measurement sample.
As the biological sample, for example, urine or whole blood is used.
In addition, by measuring the concentration of 8-hydroxydeoxyguanosine (8-OHdG) in a biological sample, it is possible to know the state of oxidative damage to guanosine, which is a constituent of DNA. The accumulation of oxidation is related to the concentration of 8-hydroxydeoxyguanosine. At the same time, the concentration of an analyte different from the 8-hydroxydeoxyguanosine having a different oxidizing action may be measured, and the oxidative stress state may be comprehensively determined based on both measurement results.
In this case, another analyte may be selected from the group consisting of, for example, chromogranin A, amylase, oxygen radical, biopyrine, thioredoxin, MDA-LDL, blood oxidized α1 antitrypsin, oxidized LDL receptor LOX-1.
Specifically, for example, 8-hydroxydeoxyguanosine present in a biological sample sample is reacted with a reactive substance that specifically reacts with 8-hydroxydeoxyguanosine, and the concentration of the substance produced or consumed by the reaction Can be measured electrically or optically to determine the concentration of 8-hydroxydeoxyguanosine in a biological sample.
The oxidative stress state may be determined by comparing the concentration of 8-hydroxydeoxyguanosine present in the biological sample with the concentration of deoxyguanosine present in the biological sample without oxidative damage. Further, the oxidative stress state may be determined by adding a known concentration of deoxyguanosine to a biological sample sample and comparing the concentration of 8-hydroxydeoxyguanosine and the concentration of deoxyguanosine before and after the addition of deoxyguanosine.
An oxidative stress state determination system according to the present invention includes a sample introduction unit that introduces a biological sample, and an 8-hydroxydeoxyguanosine concentration that measures the concentration of 8-hydroxydeoxyguanosine in the biological sample introduced from the sample introduction unit. The information processing apparatus includes a measurement unit and a determination processing unit that inputs information measured by the concentration measurement unit and determines an oxidative stress state in the body from the 8-hydroxydeoxyguanosine concentration.
For example, urine can be used as the biological sample.
The 8-hydroxydeoxyguanosine concentration measuring unit may be configured to allow a biological sample sample to react with a reactant that specifically reacts with 8-hydroxydeoxyguanosine. In this case, the 8-hydroxydeoxyguanosine concentration measuring unit may include the reactive substance in advance, or may include a reactive substance supply unit that supplies the reactive substance from the outside of the measuring unit.
The reactant that specifically reacts with 8-hydroxydeoxyguanosine may be a reagent that causes an antigen-antibody reaction with an anti-8-hydroxydeoxyguanosine antibody or a reagent that causes a binding reaction with an aptamer.
The 8-hydroxydeoxyguanosine concentration measuring unit may be configured to be able to electrically or optically measure the concentration of a substance produced or consumed by reaction with the reactant.
The means for optically measuring in the 8-hydroxydeoxyguanosine concentration measuring unit is, for example, a spectrophotometer. In this case, the spectrophotometer can be used for measuring at least one of the reflectance, fluorescence, absorbance, transmittance, or evanescent wave of the sample in the 8-hydroxydeoxyguanosine concentration measuring unit.
The means for electrically measuring in the 8-hydroxydeoxyguanosine concentration measuring unit is, for example, an electrochemical sensor. In this case, the electrochemical sensor can be used to measure at least one of a current value, a voltage value, an electric resistance value, and an impedance of the sample in the 8-hydroxydeoxyguanosine concentration measuring unit.
The 8-hydroxydeoxyguanosine concentration measurement unit includes a liquid-permeable buffer layer containing a buffer solution, a reaction layer containing the reactant, and a biological sample sample after passing through the liquid-permeable buffer layer. A support member that supports the liquid-permeable buffer layer and the reaction layer may be provided so as to reach the reaction layer.
The 8-hydroxydeoxyguanosine concentration measuring unit may further include at least one additional layer in liquid contact with the buffer layer. Specifically, the additional layer may be at least one of a blood cell separation layer, a radiation blocking layer, an interference removal layer, a dialysis layer, or a filtration layer.
The support member includes a biological sample sample introduction hole, the support member sandwiches the buffer solution layer and the reaction layer, and a biological sample sample is supplied from the sample introduction hole to the buffer solution layer and the reaction layer. Can be configured.
The 8-hydroxydeoxyguanosine concentration measurement unit includes a dye layer that holds a dye, and the buffer layer may be disposed on the dye layer or juxtaposed with the dye layer.
本発明に係る酸化ストレス状態判定方法は、生体試料サンプル中の8-ヒドロキシデオキシグアノシン(8-OHdG)の濃度を、生体試料を採取した直後の早い時間で測定し、測定した8-ヒドロキシデオキシグアノシンの濃度に基づいて、濃度が濃ければ濃いほど酸化ストレス状態が高いと判定し、濃度が低ければ低いほど酸化ストレス状態が低いと判定するので、未病の状態を事前に検知することが可能になり、より早い治療を行うことができるようになる。
また、本発明に係る酸化ストレス状態判定システムは、生体試料サンプルを導入するサンプル導入部と、サンプル導入部から導入された生体試料サンプル中の8-ヒドロキシデオキシグアノシンの濃度を測定する8-ヒドロキシデオキシグアノシン濃度測定部と、前記濃度測定部において測定された情報を入力し、8-ヒドロキシデオキシグアノシン濃度から体内の酸化ストレス状態を判定する判定処理部とを備え、前記判定部において、生体試液中の8-ヒドロキシデオキシグアノシン濃度が濃ければ濃いほど酸化ストレス状態が高いと判定し、濃度が低ければ低いほど酸化ストレス状態が低いと判定するので、未病の状態を事前に検知することが可能になり、病気になる前のより早い段階で生活習慣を見直すきっかけを作ることが出来、治療が必要な病気になることを未然に防ぐ効果をもつことができる。また、何らかの自覚症状が出始めた段階で、病院に行き医師に相談するときにでも、スクリーニングの検査により過去の酸化ストレス状態を判断することが出来、医師による適切な指導と治療指針を行うことができるようになる。なお、尿中の8-ヒドロキシデオキシグアノシンの濃度の境界値は、40ng/mLであり、この境界値を健常な人では余程の濃縮された尿サンプルでなければ超えることはなく、4ng/mL〜10ng/mLの変動、推移を個々人でモニターすることで酸化ストレス状態を把握することが出来る。
The method for determining oxidative stress state according to the present invention comprises measuring 8-hydroxydeoxyguanosine (8-OHdG) concentration in a biological sample sample at an early time immediately after collecting the biological sample, and measuring the measured 8-hydroxydeoxyguanosine. The higher the concentration, the higher the oxidative stress state, and the lower the concentration, the lower the oxidative stress state, so it is possible to detect a disease-free state in advance. It becomes possible to perform faster treatment.
The oxidative stress state determination system according to the present invention includes a sample introduction unit for introducing a biological sample, and an 8-hydroxydeoxyguanosine concentration in the biological sample introduced from the sample introduction unit. A guanosine concentration measurement unit, and a determination processing unit that inputs information measured in the concentration measurement unit and determines an oxidative stress state in the body from the 8-hydroxydeoxyguanosine concentration. In the determination unit, The higher the 8-hydroxydeoxyguanosine concentration, the higher the oxidative stress state, and the lower the concentration, the lower the oxidative stress state, so it is possible to detect an unaffected state in advance. , Can make an opportunity to review lifestyle at an earlier stage before getting sick, need treatment It is possible to have a prevent effect to become sick. In addition, when any subjective symptom begins to appear, even when going to the hospital and consulting a doctor, it is possible to determine the past oxidative stress state by screening tests, and to give appropriate guidance and treatment guidelines by the doctor. Will be able to. The boundary value of 8-hydroxydeoxyguanosine concentration in urine is 40 ng / mL, and this threshold value is not exceeded unless it is an excessively concentrated urine sample in healthy people, and it is 4 ng / mL. It is possible to grasp the oxidative stress state by monitoring the fluctuation and transition of -10ng / mL by individual.
以下、添付図面に示した一実施例を参照して、本発明に係る酸化ストレス状態判定方法を実施するための酸化ストレス状態判定システムの実施の形態を説明していく。 Hereinafter, an embodiment of an oxidative stress state determination system for carrying out an oxidative stress state determination method according to the present invention will be described with reference to an embodiment shown in the accompanying drawings.
図１は酸化ストレス状態判定システムの概略ブロック図である。
この図面に示すように、酸化ストレス状態判定システムは、生体試料サンプルを導入するサンプル導入部２０、サンプル導入部から導入された生体試料サンプル中の8-ヒドロキシデオキシグアノシンの濃度を測定する8-ヒドロキシデオキシグアノシン濃度測定部２１、及び前記濃度測定部において測定された情報を入力し、8-ヒドロキシデオキシグアノシン濃度から体内の酸化ストレス状態を判定する判定処理部２２を有する。
前記生体試料サンプルを導入するサンプル導入部２０は、生体試料サンプルを吸収するような形態であってもよく、又は、生体試料サンプルを注入する形態であってもよい。前記生体試料サンプル導入部２０からは、例えば、生体試料サンプルとして尿や全血が導入され得る。
生体試料サンプルとして尿を使用する場合、前処理をせずに直接尿をサンプル導入部２０から導入させることができるが、生体試料サンプルとして全血を使用し、かつ、測定結果が色で現れる場合には、赤血球が妨害となり判別できないため、予め血清又はは血漿が分離される。
前記8-ヒドロキシデオキシグアノシン濃度測定部２１は、生体試料サンプルと、8-ヒドロキシデオキシグアノシンに特異的に反応する反応物質とを反応させることができるように構成され得る。測定結果を色で判別する場合、例えば、前記反応物質に予め金コロイド等の存在を可視化できる可視化物質を結合させておき、8-ヒドロキシデオキシグアノシンと反応した可視化物質結合反応物質の量、又は8-ヒドロキシデオキシグアノシンと反応しなかった可視化物質結合反応物質の量に基づく色を用いて判定処理部２２において酸化ストレス状態が判定され得る。
また、前記8-ヒドロキシデオキシグアノシン濃度測定部２１は、前記反応物質を予め保持していてもよく、又は、前記反応物質を測定部２１の外部から供給する反応物質供給部２１ｇを備えていてもよい。
前記8-ヒドロキシデオキシグアノシン濃度測定部２１において用いられる8-ヒドロキシデオキシグアノシンに特異的に反応する反応物質には、抗8-ヒドロキシデオキシグアノシン抗体による抗原抗体反応を生じる試薬、又はアプタマーによる結合反応を生じる試薬が用いられる。
前記8-ヒドロキシデオキシグアノシン濃度測定部２１が、前記反応物質との反応により生成又は消費される物質の濃度を電気的又は光学的に測定することができるように構成されている。
光学的に測定を行う場合には、前記8-ヒドロキシデオキシグアノシン濃度測定部２１は、分光光度計２１ａを備え、この分光光度計２１ａで8-ヒドロキシデオキシグアノシン濃度測定部２１におけるサンプルの反射率、蛍光、吸光度、透過率又はエバネッセント波の少なくとも一つを測定する。
電気的に測定を行う場合には、前記8-ヒドロキシデオキシグアノシン濃度測定部２１は、電気化学センサ２１ｂを備え、電気化学センサが、8-ヒドロキシデオキシグアノシン濃度測定部におけるサンプルの電流値、電圧値、電気抵抗値又はインピーダンスの少なくとも一つを測定する。
前記8-ヒドロキシデオキシグアノシン濃度測定部２１は、緩衝液を含有する液透過性緩衝液層２１ｃと、前記反応物質を含有する反応層２１ｄと、生体試料サンプルが、前記液透過性緩衝液層２１ｃを通過した後に前記反応層２１ｄに至るように前記液透過性緩衝液層２１ｃ及び前記反応層２１ｄを支持する支持部材２１ｅを有する。
また、試料サンプルの種類や測定方法等によって必要に応じて、前記8-ヒドロキシデオキシグアノシン濃度測定部２１は、前記緩衝液層２１ｃと液接触する少なくとも一つの付加的な層（図示せず）を有することができる。具体的には、前記付加的な層は、血球分離層、放射線遮断層、干渉除去層、透析層又は濾過層の少なくとも一つである。
前記支持部材２１ｅは、サンプル導入部２０を介して生体試料サンプルが導入される生体試料サンプル導入孔２１ｆを備え、かつ、前記支持部材２１ｅは、前記緩衝液層２１ｃ及び前記反応層２１ｄを挟み込み、前記サンプル導入孔２０から生体試料サンプルが前記緩衝液層２１ｃ及び前記反応層２１ｄに供給されるように構成されている。
前記反応層２１ｄでは、生体試料サンプル中の測定対象物と前記反応物質が反応し、それにより生成もしくは消費した物質により色が変化するように構成されている。
上記したように構成された前記8-ヒドロキシデオキシグアノシン濃度測定部２１において測定された測定結果は、判定処理部２２に送られ、判定処理部２２において、生体試料サンプル中の8-ヒドロキシデオキシグアノシンの濃度に基づいて酸化ストレス状態の判定を行う。具体的には、生体試料サンプル中の8-ヒドロキシデオキシグアノシンの濃度が高ければ高いほど、酸化ストレス状態が高いと判定し、同濃度が低ければ低い程、酸化ストレス状態が低いと判定する。
FIG. 1 is a schematic block diagram of an oxidative stress state determination system.
As shown in this drawing, the oxidative stress state determination system includes a sample introduction unit 20 for introducing a biological sample, and an 8-hydroxydeoxyguanosine concentration in the biological sample introduced from the sample introduction unit. A deoxyguanosine concentration measuring unit 21 and a determination processing unit 22 for inputting information measured by the concentration measuring unit and determining an oxidative stress state in the body from the 8-hydroxydeoxyguanosine concentration are included.
The sample introduction unit 20 for introducing the biological sample sample may be configured to absorb the biological sample sample or may be configured to inject the biological sample sample. From the biological sample sample introduction unit 20, for example, urine or whole blood can be introduced as a biological sample sample.
When urine is used as a biological sample, urine can be directly introduced from the sample introduction unit 20 without pretreatment, but when whole blood is used as the biological sample and the measurement result appears in color In this case, erythrocytes are obstructed and cannot be discriminated, so that serum or plasma is separated in advance.
The 8-hydroxydeoxyguanosine concentration measuring unit 21 may be configured to allow a biological sample sample to react with a reactant that specifically reacts with 8-hydroxydeoxyguanosine. When discriminating the measurement result by color, for example, a visualizing substance capable of visualizing the presence of colloidal gold or the like is bound in advance to the reactive substance, and the amount of the visualizing substance-bound reactive substance reacted with 8-hydroxydeoxyguanosine, or 8 The oxidative stress state can be determined in the determination processing unit 22 using a color based on the amount of the visualization substance binding reactant that has not reacted with -hydroxydeoxyguanosine.
The 8-hydroxydeoxyguanosine concentration measuring unit 21 may hold the reactant in advance, or may include a reactant supply unit 21g that supplies the reactant from outside the measuring unit 21. Good.
The reactant that specifically reacts with 8-hydroxydeoxyguanosine used in the 8-hydroxydeoxyguanosine concentration measuring unit 21 includes a reagent that causes an antigen-antibody reaction with an anti-8-hydroxydeoxyguanosine antibody, or a binding reaction with an aptamer. The resulting reagent is used.
The 8-hydroxydeoxyguanosine concentration measuring unit 21 is configured to be able to electrically or optically measure the concentration of a substance generated or consumed by reaction with the reactant.
When optically measuring, the 8-hydroxydeoxyguanosine concentration measuring unit 21 includes a spectrophotometer 21a, and the spectrophotometer 21a uses the reflectance of the sample in the 8-hydroxydeoxyguanosine concentration measuring unit 21; At least one of fluorescence, absorbance, transmittance, or evanescent wave is measured.
When the measurement is performed electrically, the 8-hydroxydeoxyguanosine concentration measurement unit 21 includes an electrochemical sensor 21b, and the electrochemical sensor has a current value and a voltage value of a sample in the 8-hydroxydeoxyguanosine concentration measurement unit. Measure at least one of electrical resistance or impedance.
The 8-hydroxydeoxyguanosine concentration measuring unit 21 includes a liquid-permeable buffer layer 21c containing a buffer solution, a reaction layer 21d containing the reactant, and a biological sample sample containing the liquid-permeable buffer layer 21c. And a support member 21e for supporting the liquid permeable buffer solution layer 21c and the reaction layer 21d so as to reach the reaction layer 21d.
In addition, the 8-hydroxydeoxyguanosine concentration measuring unit 21 may include at least one additional layer (not shown) in liquid contact with the buffer layer 21c as necessary depending on the type of sample sample, the measurement method, and the like. Can have. Specifically, the additional layer is at least one of a blood cell separation layer, a radiation blocking layer, an interference removal layer, a dialysis layer, or a filtration layer.
The support member 21e includes a biological sample sample introduction hole 21f through which a biological sample sample is introduced via the sample introduction unit 20, and the support member 21e sandwiches the buffer layer 21c and the reaction layer 21d, A biological sample is supplied from the sample introduction hole 20 to the buffer solution layer 21c and the reaction layer 21d.
The reaction layer 21d is configured such that the measurement object in the biological sample sample reacts with the reactant and the color changes depending on the substance generated or consumed.
The measurement result measured by the 8-hydroxydeoxyguanosine concentration measuring unit 21 configured as described above is sent to the determination processing unit 22, and the determination processing unit 22 determines the 8-hydroxydeoxyguanosine in the biological sample sample. The oxidative stress state is determined based on the concentration. Specifically, the higher the 8-hydroxydeoxyguanosine concentration in the biological sample is determined, the higher the oxidative stress state, and the lower the concentration, the lower the oxidative stress state.
次に、本発明における酸化ストレス状態判定方法で使用可能なイムノクロマトセンサの実施例を説明していく。
図２は、イムノクロマトセンサの分解展開図、図３は図２に示したイムノクロマトセンサの上面カバー以外の部分を組立てた部分分解展開図、図４は図２に示したイムノクロマトセンサの上面図を各々示している。
図中、符号１は土台となるベース基板を示しており、このベース基板１の上に、テストストリップ２及び試液吸収部３が設けられている。
テストストリップ２は、試液吸収部３に部分的に重ねて配置された試液展開層４と、前記試液展開層４に部分的に重ねて配置された可視化物質結合抗体保持層５と、可視化物質結合抗体保持層５に部分的に重ねて配置された展開層６とから成る。
可視化物質結合抗体保持層５には、測定すべき物質と特異的に反応する抗体に、その抗体の存在を可視化できる可視化物質を結合させた可視化物質結合抗体が保持されている。具体的には、例えば、測定すべき物質が8-ヒドロキシデオキシグアノシンの場合、その抗体は、8OHdG1B1(独立行政法人 産業技術総合研究所 特許生物寄託センター 受託番号FERM P-20122)（本願の出願人が特開2006-056859にて既に提案している抗体）であり、可視化物質としては金コロイドが挙げられる。この可視化物質結合抗体保持層５に保持された可視化物質結合抗体は、試液展開層４を介して試液が導入されると試液中に溶出する。このため、可視化物質結合抗体保持層５は、使用前（即ち、試液が通過する前）は、可視化物質（例えば、金コロイド）によって色付いているが、使用後、即ち、試液が正常に通過した後は、可視化物質が試液中に溶出してしまうため色が無くなる。具体的には、可視化物質結合抗体保持層５が白いメンブレンから成り、可視化物質が金コロイドである場合には、可視化物質結合抗体保持層５は、使用前は赤色であるが、使用後は白色になる。
展開層６は、前記可視化物質結合抗体保持層５に保持された可視化物質結合抗体と結合して、同抗体を捕捉する固定化抗原物質が固定された判定部７を有する。この判定部７は、展開層６において測定すべき物質と可視化物質結合抗体とが充分に抗体抗原反応することができるように、可視化物質結合抗体保持層５から充分に離れた位置に設けられている。
Next, an example of an immunochromatographic sensor that can be used in the oxidative stress state determination method of the present invention will be described.
2 is an exploded development view of the immunochromatographic sensor, FIG. 3 is a partial exploded development view in which parts other than the top cover of the immunochromatographic sensor shown in FIG. 2 are assembled, and FIG. 4 is a top view of the immunochromatographic sensor shown in FIG. Show.
In the figure, reference numeral 1 denotes a base substrate that serves as a base, and a test strip 2 and a reagent solution absorbing portion 3 are provided on the base substrate 1.
The test strip 2 includes a reagent solution developing layer 4 that is partially overlapped with the reagent solution absorbing portion 3, a visualization substance binding antibody holding layer 5 that is partially overlapped with the reagent solution developing layer 4, and a visualization substance binding. And a spreading layer 6 disposed partially overlapping the antibody holding layer 5.
The visualization substance-bound antibody holding layer 5 holds a visualization substance-binding antibody in which a visualization substance capable of visualizing the presence of the antibody is bound to an antibody that specifically reacts with the substance to be measured. Specifically, for example, when the substance to be measured is 8-hydroxydeoxyguanosine, the antibody is 8OHdG1B1 (National Institute of Advanced Industrial Science and Technology, Patent Biodeposition Center Accession No.FERM P-20122) (the applicant of the present application). Is an antibody already proposed in Japanese Patent Application Laid-Open No. 2006-056859), and examples of the visualization substance include colloidal gold. The visualized substance-bound antibody retained in the visualized substance-bound antibody retaining layer 5 is eluted in the reagent when the reagent is introduced through the reagent developing layer 4. Therefore, the visualization substance-bound antibody holding layer 5 is colored by the visualization substance (for example, gold colloid) before use (that is, before the sample solution passes), but after use, that is, the sample solution has passed normally. After that, since the visualization substance is eluted in the test solution, the color disappears. Specifically, when the visualization substance binding antibody holding layer 5 is made of a white membrane and the visualization substance is a gold colloid, the visualization substance binding antibody holding layer 5 is red before use but is white after use. become.
The development layer 6 includes a determination unit 7 to which an immobilized antigen substance that binds to the visualization substance-bound antibody retained in the visualization substance-bound antibody retention layer 5 and captures the antibody is immobilized. The determination unit 7 is provided at a position sufficiently separated from the visualization substance-bound antibody holding layer 5 so that the substance to be measured in the development layer 6 and the visualization substance-bound antibody can sufficiently react with each other. Yes.
上記したように構成されたテストストリップ２及び試液吸収部３はベース基板１上に配置され、さらに基板１にカバー１０が取り付けられる。
この実施例では、カバー１０は、ベース基板１における試液吸収部３以外の部分を全て覆うように寸法決めされており、かつ、テストストリップ２に相当する部分には確認用窓１１が形成されている。
この確認用窓１１は、可視化物質結合抗体保持層５の一部と、展開層６における判定部７が露出するように形成されている。
また、カバー１０における判定部７に相当する部分には、判定部７を挟むように色調表１２が設けられている。この実施例では、図３における判定部７の左側に測定すべき物質の量が少ない時の色（即ち、濃い色）が、右側に測定すべき物質の量が多い時の色（即ち、薄い色）が印刷されている。
The test strip 2 and the test solution absorber 3 configured as described above are disposed on the base substrate 1, and a cover 10 is attached to the substrate 1.
In this embodiment, the cover 10 is dimensioned so as to cover all portions of the base substrate 1 other than the reagent solution absorbing portion 3, and a confirmation window 11 is formed in a portion corresponding to the test strip 2. Yes.
This confirmation window 11 is formed so that a part of the visualization substance binding antibody holding layer 5 and the determination part 7 in the development layer 6 are exposed.
A color tone table 12 is provided at a portion corresponding to the determination unit 7 in the cover 10 so as to sandwich the determination unit 7. In this embodiment, the color when the amount of the substance to be measured is small on the left side of the determination unit 7 in FIG. 3 (that is, the dark color) is the color when the amount of the substance to be measured is large on the right side (that is, the color is light). Color) is printed.
次に、上記したように構成されたイムノクロマトセンサの作用を説明していく。
図５は、上記したイムノクロマトセンサの作用を示す図であり、（ａ）は使用前の状態を、（ｂ）は使用後の状態を各々示している。
この説明では、測定すべき物質は、尿中に含まれている8-ヒドロキシデオキシグアノシンの量であり、従って、抗体は、8-ヒドロキシデオキシグアノシンに特異的に反応する抗体（8OHdG1B1(独立行政法人 産業技術総合研究所 特許生物寄託センター 受託番号FERM P-20122)）であり、可視化物質は金コロイドである。また、判定層に固定されている固定化物質は8-ヒドロキシデオキシグアノシン(8OHdG)である。
使用者が、尿を試液吸収部３にかけると、尿は試液吸収部３に吸収される。試液吸収部３に吸収した尿は、毛細管現象により試液展開層４に入り、試液展開層４の幅全体に広がりながら可視化物質結合抗体保持層５に進む。そして、尿が可視化物質結合抗体保持層５を通過する時に、尿中に、同層５に保持された金コロイドが結合した抗体（8OHdG1B1(独立行政法人 産業技術総合研究所 特許生物寄託センター 受託番号FERM P-20122)）が溶出する。金コロイド結合抗体が溶出した尿は、そのまま展開層６へ進み、同層６で8-ヒドロキシデオキシグアノシンと金コロイド結合抗体とが抗体抗原反応して、尿中にある8-ヒドロキシデオキシグアノシンに金コロイド結合抗体が結合する。展開層６に入った尿は、さらに、毛細管現象により展開層６を判定部７に向かって進む。
尿が判定部７を通過する時に、既に8-ヒドロキシデオキシグアノシンと結合した金コロイド結合抗体は判定部７を通過するが、8-ヒドロキシデオキシグアノシンと結合していない金コロイド結合抗体は判定部７に固定された固定化物質と結合して判定部７に留まる。このため、判定部７は固定化物質と結合した金コロイド結合抗体の金コロイドの色の作用で赤く見える。
尿中に含まれている8-ヒドロキシデオキシグアノシンの量が多ければ多いほど、8-ヒドロキシデオキシグアノシンと結合しない金コロイド結合抗体の量は少なくなるので判定部７の色は薄くなり、逆に、尿中に含まれている8-ヒドロキシデオキシグアノシンの量が少なければ少ないほど、8-ヒドロキシデオキシグアノシンと結合しない金コロイド結合抗体の量が多くなるので判定部７の色は濃くなる。
使用者は、判定部７の両隣にある色調表１２の色と判定部７の色とを比較して、尿中に含まれている8-ヒドロキシデオキシグアノシンの量が多いか否かを判断する。
ところで、上記したように、このイムノクロマトセンサはカバー１０の窓部１１が、可視化物質結合抗体保持層５の一部が露出するように形成されている。このため、使用者は、窓部１１から見える可視化物質結合抗体保持層５の色の変化によって、尿が展開層６に入っているか否かを確認することができる。具体的には、可視化物質結合抗体保持層５は、使用前は、そこに保持されている可視化物質結合抗体の可視化物質、ここでは金コロイドの作用で色付いている。そして、可視化物質結合抗体保持層５は試液、ここでは尿が通過すると、可視化物質結合抗体が試液中に溶出するため同層５の色はなくなる。従って、試液導入部３から導入した尿が、何らかの原因で展開層６に進まなければ可視化物質結合抗体保持層５の色は変わらないが、試液導入部３から導入した尿が、正確に展開層６に進めば可視化物質結合抗体保持層５の色が変わる。このため、使用者は、窓部１１から見える可視化物質結合抗体保持層５の色によって試液が正確に展開層６に進んでいるか否かを確認することができるため、判定結果を間違えて認識することがない。
Next, the operation of the immunochromatographic sensor configured as described above will be described.
FIGS. 5A and 5B are diagrams showing the operation of the immunochromatographic sensor described above. FIG. 5A shows a state before use, and FIG. 5B shows a state after use.
In this description, the substance to be measured is the amount of 8-hydroxydeoxyguanosine contained in the urine. Therefore, the antibody is an antibody (8OHdG1B1 (independent administrative agency) that specifically reacts with 8-hydroxydeoxyguanosine. The National Institute of Advanced Industrial Science and Technology, Patent Biological Deposit Center, Accession Number FERM P-20122)), and the visualization material is colloidal gold. The immobilization substance immobilized on the determination layer is 8-hydroxydeoxyguanosine (8OHdG).
When the user puts urine on the sample solution absorption unit 3, the urine is absorbed by the sample solution absorption unit 3. The urine absorbed in the test solution absorbing part 3 enters the test solution developing layer 4 by capillary action and proceeds to the visualization substance-binding antibody holding layer 5 while spreading over the entire width of the test solution developing layer 4. When the urine passes through the visualization substance-binding antibody holding layer 5, the antibody (8OHdG1B1 (Independent Administrative Institution, National Institute of Advanced Industrial Science and Technology, Patent Biological Depositary Accession No.) FERM P-20122)) elutes. The urine from which the gold colloid-binding antibody is eluted proceeds to the developing layer 6 as it is, and 8-hydroxydeoxyguanosine and the gold colloid-binding antibody react with each other in the same layer 6 to form gold into 8-hydroxydeoxyguanosine in the urine. Colloid binding antibody binds. The urine that has entered the development layer 6 further proceeds through the development layer 6 toward the determination unit 7 by capillary action.
When the urine passes through the determination unit 7, the gold colloid-binding antibody already bound to 8-hydroxydeoxyguanosine passes the determination unit 7, but the gold colloid-binding antibody not bonded to 8-hydroxydeoxyguanosine determines the determination unit 7. It remains in the determination unit 7 by combining with the immobilized substance fixed on the surface. For this reason, the determination unit 7 looks red due to the effect of the colloidal gold color of the colloidal gold antibody bound to the immobilized substance.
The greater the amount of 8-hydroxydeoxyguanosine contained in the urine, the smaller the amount of colloidal gold-bound antibody that does not bind to 8-hydroxydeoxyguanosine, so the color of the determination unit 7 becomes lighter. The smaller the amount of 8-hydroxydeoxyguanosine contained in the urine, the greater the amount of colloidal gold-bound antibody that does not bind to 8-hydroxydeoxyguanosine, so the color of the determination unit 7 becomes darker.
The user compares the color of the color tone table 12 on both sides of the determination unit 7 with the color of the determination unit 7 to determine whether the amount of 8-hydroxydeoxyguanosine contained in the urine is large. .
By the way, as described above, in this immunochromatographic sensor, the window portion 11 of the cover 10 is formed so that a part of the visualization substance binding antibody holding layer 5 is exposed. For this reason, the user can confirm whether or not urine is in the spreading layer 6 by a change in the color of the visualization substance binding antibody holding layer 5 visible from the window portion 11. Specifically, the visualized substance-bound antibody holding layer 5 is colored by the action of the visualized substance of the visualized substance-bound antibody held therein, here, colloidal gold, before use. The visualization substance-binding antibody holding layer 5 disappears when the test solution, here urine passes, because the visualization substance-binding antibody is eluted in the test solution. Therefore, the color of the visualization substance-bound antibody retaining layer 5 does not change unless the urine introduced from the reagent introduction part 3 proceeds to the development layer 6 for some reason, but the urine introduced from the reagent introduction part 3 does not accurately If it advances to 6, the color of the visualization substance binding antibody holding layer 5 will change. For this reason, since the user can confirm whether or not the test solution has advanced to the developing layer 6 accurately based on the color of the visualization substance-binding antibody holding layer 5 that can be seen from the window portion 11, the determination result is mistakenly recognized. There is nothing.
上記したイムノクロマトセンサの実施例では、使用者が、判定部７の両隣にある色調表１２の色と判定部７の色とを比較して、尿中に含まれている8-ヒドロキシデオキシグアノシンの量が多いか否かを判断し、尿中に含まれている8-ヒドロキシデオキシグアノシンの量が多ければ多いほど酸化ストレス状態が高く、同濃度が低ければ酸化ストレス状態が低いと判定できるように構成した例を挙げているが、生体試料中の8-ヒドロキシデオキシグアノシンの濃度に基づく酸化ストレス状態の判定は、本実施例に限定されることなく、例えば、判定部７の色を光学的に測定する手段（例えば、分光光度計）で測定して、その測定結果に基づいて、予め、測定結果に応じた酸化ストレス状態の高さを記憶するか、又は測定結果に応じた酸化ストレス状態の算出方法を記憶した判定処理手段を用いて酸化ストレス状態を判定してもよい。 In the embodiment of the immunochromatographic sensor described above, the user compares the color of the color tone table 12 on both sides of the determination unit 7 with the color of the determination unit 7 to determine the 8-hydroxydeoxyguanosine contained in the urine. Judging whether the amount is high, the higher the amount of 8-hydroxydeoxyguanosine contained in the urine, the higher the oxidative stress state, and the lower the concentration, the lower the oxidative stress state Although the example which comprised was given, determination of the oxidative stress state based on the density | concentration of 8-hydroxydeoxyguanosine in a biological sample is not limited to a present Example, For example, the color of the determination part 7 is optically determined. Measured with a measuring means (for example, a spectrophotometer), and based on the measurement result, the height of the oxidative stress state corresponding to the measurement result is stored in advance, or the oxidative stress state according to the measurement result The method of calculating may determine the oxidative stress status using the determination processing means for storing.
１ ベース基板
２ テストストリップ
３ 試液吸収部
４ 試液展開層
５ 可視化物質結合抗体保持層
６ 展開層
７ 判定部
１０ カバー
１１ 窓部
１２ 色調表
２０ サンプル導入部
２１ 8-ヒドロキシデオキシグアノシン濃度測定部
２１ａ 分光光度計
２１ｂ 電気化学センサ
２１ｃ 液透過性緩衝液層
２１ｄ 反応層
２１ｅ 支持部材
２１ｆ サンプル導入孔
２１ｇ 反応物質供給部
２２ 判定処理部
DESCRIPTION OF SYMBOLS 1 Base substrate 2 Test strip 3 Reagent absorption part 4 Reagent development layer 5 Visualization substance binding antibody holding layer 6 Development layer 7 Judgment part 10 Cover 11 Window part 12 Color tone table
20 Sample introduction unit 21 8-hydroxydeoxyguanosine concentration measurement unit 21a Spectrophotometer 21b Electrochemical sensor 21c Liquid permeable buffer layer 21d Reaction layer 21e Support member 21f Sample introduction hole 21g Reactant supply unit 22 Determination processing unit
Claims (26)
測定した8-ヒドロキシデオキシグアノシンの濃度に基づいて、体内の酸化ストレス状態を判定する
ことを特徴とする酸化ストレス状態判定方法。 Measure the concentration of 8-hydroxydeoxyguanosine (8-OHdG) in the biological sample,
An oxidative stress state determination method characterized by determining an oxidative stress state in the body based on the measured concentration of 8-hydroxydeoxyguanosine.
ことを特徴とする請求項１に記載の酸化ストレス状態判定方法。 The method for determining an oxidative stress state according to claim 1, wherein the concentration of 8-hydroxydeoxyguanosine in a biological sample sample that has not been pretreated is measured.
ことを特徴とする請求項１に記載の酸化ストレス状態判定方法。 The oxidative stress state determination method according to claim 1, wherein the biological sample is mixed with the measurement sample, and the concentration of 8-hydroxydeoxyguanosine in the measurement sample is measured in one test.
ことを特徴とする請求項１〜３の何れか一項に記載の酸化ストレス状態判定方法。 The oxidative stress state determination method according to claim 1, wherein the biological sample is urine.
ことを特徴とする請求項１〜４の何れか一項に記載の酸化ストレス状態判定方法。 The oxidative stress state determination method according to any one of claims 1 to 4, wherein the biological sample is whole blood.
ことを特徴とする請求項１〜５の何れか一項に記載の酸化ストレス状態判定方法。 The oxidation according to any one of claims 1 to 5, wherein a concentration of an analyte different from the 8-hydroxydeoxyguanosine in the biological sample is further measured as an index of an oxidative stress state. Stress state determination method.
クロモグラニンＡ、アミラーゼ、酸素ラジカル、バイオピリン、チオレドキシン、ＭＤＡ−ＬＤＬ、血中酸化型α１アンチトリプシン、酸化ＬＤＬ受容体ＬＯＸ−１からなる群から選択される
ことを特徴とする請求項６に記載の酸化ストレス状態判定方法。 Said another analyte is
The oxidation according to claim 6, which is selected from the group consisting of chromogranin A, amylase, oxygen radical, biopyrine, thioredoxin, MDA-LDL, blood oxidized α1 antitrypsin, and oxidized LDL receptor LOX-1. Stress state determination method.
該8-ヒドロキシデオキシグアノシンに特異的に反応する反応物質とを反応させ、
反応により生成又は消費される物質の濃度を電気的又は光学的に測定することにより、
生体試料サンプル中の8-ヒドロキシデオキシグアノシンの濃度を測定する
ことを特徴とする請求項１〜７の何れか一項に記載の酸化ストレス状態判定方法。 8-hydroxydeoxyguanosine present in the biological sample;
Reacting with a reactant that specifically reacts with the 8-hydroxydeoxyguanosine,
By measuring the concentration of substances produced or consumed by the reaction electrically or optically,
The method for determining an oxidative stress state according to any one of claims 1 to 7, wherein the concentration of 8-hydroxydeoxyguanosine in the biological sample is measured.
酸化損傷を受けることなく生体試料サンプル中に存在する、もしくはあらかじめ生体試料サンプル中に投入したデオキシグアノシンの濃度と
を比較することによって酸化ストレス状態の判定を行う
ことを特徴とする請求項１〜８の何れか一項に記載の酸化ストレス状態判定方法。 The concentration of 8-hydroxydeoxyguanosine present in the biological sample;
The oxidative stress state is determined by comparing the concentration of deoxyguanosine that is present in a biological sample sample without being damaged by oxidization, or that has been previously introduced into the biological sample sample. The oxidative stress state determination method according to any one of the above.
サンプル導入部から導入された生体試料サンプル中の8-ヒドロキシデオキシグアノシンの濃度を測定する8-ヒドロキシデオキシグアノシン濃度測定部と、
前記濃度測定部において測定された情報を入力し、8-ヒドロキシデオキシグアノシン濃度から体内の酸化ストレス状態を判定する判定処理部と
を備えていることを特徴とする酸化ストレス状態判定システム。 A sample introduction part for introducing a biological sample, and
An 8-hydroxydeoxyguanosine concentration measuring unit for measuring the concentration of 8-hydroxydeoxyguanosine in the biological sample sample introduced from the sample introduction unit;
An oxidative stress state determination system comprising: a determination processing unit that inputs information measured by the concentration measurement unit and determines an oxidative stress state in the body from an 8-hydroxydeoxyguanosine concentration.
ことを特徴とする請求項１０に記載の酸化ストレス状態判定システム。 The oxidative stress state determination system according to claim 10, wherein the biological sample is urine.
ことを特徴とする請求項１０又は１１に記載の酸化ストレス状態判定システム。 The said 8-hydroxydeoxyguanosine density | concentration measurement part is comprised so that a biological sample sample and the reactive substance which reacts specifically with 8-hydroxydeoxyguanosine can be made to react. Or the oxidative stress state determination system according to 11.
ことを特徴とする請求項１２に記載の酸化ストレス状態判定システム。 The oxidative stress state determination system according to claim 12, wherein the 8-hydroxydeoxyguanosine concentration measuring unit includes the reactive substance in advance.
ことを特徴とする請求項１２に記載の酸化ストレス状態判定システム。 The oxidative stress state determination system according to claim 12, wherein the 8-hydroxydeoxyguanosine concentration measurement unit includes a reaction material supply unit that supplies the reaction material from outside the measurement unit.
抗8-ヒドロキシデオキシグアノシン抗体による抗原抗体反応を生じる試薬、又は
アプタマーによる結合反応を生じる試薬である
ことを特徴とする請求項１２〜１４の何れか一項に記載の酸化ストレス状態判定システム。 A reactive substance that specifically reacts with 8-hydroxydeoxyguanosine
The oxidative stress state determination system according to any one of claims 12 to 14, which is a reagent that causes an antigen-antibody reaction with an anti-8-hydroxydeoxyguanosine antibody or a reagent that causes a binding reaction with an aptamer.
ことを特徴とする請求項１２〜１５の何れか一項に記載の酸化ストレス状態判定システム。 The 8-hydroxydeoxyguanosine concentration measurement unit is configured to be able to electrically or optically measure the concentration of a substance produced or consumed by reaction with the reactant. Item 16. The oxidative stress state determination system according to any one of Items 12 to 15.
ことを特徴とする請求項１６に記載の酸化ストレス状態判定システム。 The oxidative stress state determination system according to claim 16, wherein the optically measuring means in the 8-hydroxydeoxyguanosine concentration measuring unit is a visual or spectrophotometer.
ことを特徴とする請求項１７に記載の酸化ストレス状態判定システム。 The spectrophotometer is used to measure at least one of reflectance, fluorescence, absorbance, transmittance, or evanescent wave of a sample in an 8-hydroxydeoxyguanosine concentration measurement unit. Oxidative stress state determination system.
ことを特徴とする請求項１６に記載の酸化ストレス状態判定システム。 The oxidative stress state determination system according to claim 16, wherein the means for electrically measuring in the 8-hydroxydeoxyguanosine concentration measuring unit is an electrochemical sensor.
ことを特徴とする請求項１９に記載の酸化ストレス状態判定システム。 The electrochemical sensor is used to measure at least one of a current value, a voltage value, an electric resistance value, and an impedance of a sample in an 8-hydroxydeoxyguanosine concentration measurement unit. Oxidative stress state determination system.
緩衝液を含有する液透過性緩衝液層と、
前記反応物質を含有する反応層と、
生体試料サンプルが、前記液透過性緩衝液層を通過した後に前記反応層に至るように前記液透過性緩衝液層及び前記反応層を支持する支持部材を有する
ことを特徴とする請求項１２〜２０の何れか一項に記載の酸化ストレス状態判定システム。 The 8-hydroxydeoxyguanosine concentration measuring unit is
A liquid permeable buffer layer containing a buffer;
A reaction layer containing the reactant,
The biological sample sample has a support member that supports the liquid permeable buffer solution layer and the reaction layer so as to reach the reaction layer after passing through the liquid permeable buffer solution layer. The oxidative stress state determination system according to any one of 20.
前記緩衝液層と液接触する少なくとも一つの付加的な層をさらに含む
ことを特徴とする請求項２１に記載の酸化ストレス状態判定システム。 The 8-hydroxydeoxyguanosine concentration measuring unit is
The oxidative stress state determination system according to claim 21, further comprising at least one additional layer in liquid contact with the buffer layer.
血球分離層、
放射線遮断層、
干渉除去層、
透析層又は
濾過層
の少なくとも一つである
ことを特徴とする請求項２２に記載の酸化ストレス状態判定システム。 The additional layer is
Blood cell separation layer,
Radiation blocking layer,
Interference cancellation layer,
The oxidative stress state determination system according to claim 22, wherein the oxidative stress state determination system is at least one of a dialysis layer and a filtration layer.
前記支持部材が、前記緩衝液層及び前記反応層を挟み込み、前記サンプル導入孔から生体試料サンプルが前記緩衝液層及び前記反応層に供給されるように構成されている
ことを特徴とする請求項２１〜２３の何れか一項に記載の酸化ストレス状態判定システム。 The support member includes a biological sample sample introduction hole; and
The said support member is comprised so that the said buffer solution layer and the said reaction layer may be pinched | interposed, and a biological sample sample may be supplied to the said buffer solution layer and the said reaction layer from the said sample introduction hole. The oxidative stress state determination system according to any one of 21 to 23.
前記緩衝液層が前記色素層の上に重ねて配置される
ことを特徴とする請求項２１〜２４の何れか一項に記載の酸化ストレス状態判定システム。 The 8-hydroxydeoxyguanosine concentration measuring unit includes a dye layer for holding a dye,
The oxidative stress state determination system according to any one of claims 21 to 24, wherein the buffer solution layer is disposed so as to overlap the dye layer.
前記緩衝液層が前記色素層と並置される
ことを特徴とする請求項２１〜２４の何れか一項に記載の酸化ストレス状態判定システム。 The 8-hydroxydeoxyguanosine concentration measuring unit includes a dye layer for holding a dye,
The oxidative stress state determination system according to any one of claims 21 to 24, wherein the buffer solution layer is juxtaposed with the dye layer.
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| EP2405335A2 (en) | 2010-07-09 | 2012-01-11 | Sony Corporation | Display control apparatus and display control method, display control program, and recording medium |
| WO2019004195A1 (en) * | 2017-06-30 | 2019-01-03 | キヤノン株式会社 | Kit for chromatography, developing liquid for chromatography, and chromatography |
| JP7461037B2 (en) | 2020-07-22 | 2024-04-03 | 株式会社Resvo | Biomarkers for determining sleep disorders |
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