1269036 九、發明說明： '· 【發明所屬之技術領域】 • 本創作係一種電化學生物感測器試片，其係一種利用氧化或還原物質 與檢體中之待測物發生專一性電化學反應而產生電子轉移，配合檢測儀哭 使用，檢測出該檢體濃度之試片，且該反應區周圍係呈全面開放狀，而八 檢體待測物更易進入反應區，不受方向限制者。 【先别技術】 籲 由於居家護理之觀念曰益提升，快速、便宜、不需專業人員操作之居 家自我檢測產品日漸增多，例如血糖儀器、電子耳溫搶、電子式血壓計等， 其中血糖儀所使用之拋棄式血糖試片即是利用電化學生物感測器原理檢測 檢體中血糖濃度。 目前市面上有許多血糖試片產品，其施加檢體方式主要區分為滴落式 及定量式(毛細作时式），試片設計上祕式試片通常以塑膠網覆蓋於反 應區上避免污染，但其檢體需求量一般大於3微升，這對於老年人或資深 籲糖JC病患以採血針採集血液檢體，必須凝聚成一滴血才可以準確、足夠量 滴落至反應區，相當不方便’常造成操作上之誤差而浪費試片。 而定量式(毛細作用方式)試片之檢體需要量小於3微升町，只要少 許檢體接觸吸人口，檢體會自動吸引至反應區，但目前市售虹吸式制二 設計上不是試片尺寸過小造成細時不方便且易污染吸人口，同時吸入口 通常只有-個且方位固定，造成操作不便，況且，此種試片之需灰量小於3 微升’皆是以縮小反應區電極面積來達成，但縮小面積卻會相對造成訊號 微弱’必細麟密顧制，以觀號偵測。 1269036 請參照第1圖所示，其為美國專利號碼第5, 762, 770號，主要揭示一 種滴落式片狀電極試片’包括一電絕緣基板，其同一面上有一兩極式電極 系統；一電絕緣層覆蓋於電極系統上並界定出反應區及連接偵測器之陰陽 兩極接頭；一反應膜佈滿於反應區内，再覆蓋一塑膠網於反應區上方，最 後貼上一反應區露空之膠帶將塑膠網固定。 然而此種試片之檢體施加方式必須以滴落方式滴於反應區之上方因 此檢體滴落於反應區之方向自然受到限制’同時檢體需求量必須在9微升 以上。 睛參照第2圖所示，其為美國專利號碼第5,12〇，㈣號，主要揭示一 種毛細作用方式片狀電極試片’包括—電絕緣基板，其同—面上有一電極 系統、-電絕緣層、-反應難及位於槪麵上方之親水性上蓋板形成 檢體承載反應區空間，其中該檢體承載反應區空間須具有—個檢體吸入口 與一個氣體排出口。其找反顧之製作乃是依序於該賴基板之反應區 上塗佈缓基甲基纖維素(Carb〇Xyiflethylcellul〇se，CMC)水溶液，乾燥後形 成-親水性高分子CMC層；繼續於其上塗佈_糖氧化酵素(Gluc〇se oxidase)溶液，乾驗形成G_，再於其上魏親水性高分子-聚乙稀一 氮五酬(P〇lyvinylpyrrolid〇ne)溶液，進行第三次乾燥後形成保護膜 (PVP層）’在於其上噴覆含有電子媒介物之有機姆浮溶液，乾燥後即形 反應膜。 v 該毛細作用方式片狀電極試片之缺點在於··第一、完成反應膜需四個 /驟域CMC層用以改善反應區電極表面之疏水性；形成⑽層、親水 1269036 性保護膜PVP層及電子媒介物層，每形成一層須先乾燥，製程步驟繁瑣。 第二、施加檢體方式為接觸電極試片之尖端後，檢體會自動導入反應區中， 但只有一個位置可施加檢體。第三、試片過小取用不方便且易污染檢體吸 入π 〇 請參照第3圖所示，其為台灣公告專利號第573735號專利，其為一種 血糖測試試片，該試片包含： 一第一基板;一第一電極，係配置於第一基板上，一端為電極連接點， 另一端為電極酵素作用區； 一絕緣層’係覆蓋於第一基板上，且於電極酵素作用區具有一開口； 一酵素膜，係塗佈於絕緣層之開口處之電極酵素作用區；以及一第二 基板，係覆蓋於該血糖測試試片的最上層，於底面具有一第二電極，該第 二電極之一端為電極連接點，另一端為電極酵素作用區。 其中血糖測試試片之絕緣層還形成一狹縫以便實行虹吸作用，且於靠 近狹縫末端處形成一開孔以為虹吸作用之透氣孔。 此種結構雖可解決上述之問題，但因其開口有一定方向性，因此面積 相對縮小，且在滴入檢體亦有方向及範圍之限制。 再者，其必須在靠近狹縫末端處形成一開孔以為虹吸作用之透氣孔， 增加使用及製造上之困難。 清參照第4圖所示，其為美國專利號碼第6,258,229 β1號，其主要揭 不-纖體需求量小於丨微升之毛細作用方式電極試片，包括一電絕緣基 板，基板同一面上一三極式電極系統，一絕緣中間層以形成1]型通道，一 1269036 -· 具有一孔洞之上蓋層，然而其缺點在於檢體加入口只有一個且位於試片頂 ' 端。 •請參照第5圖所示，其為台灣專利公告第570192號專利案，其為一種 厚臈電極式生化感測試片，具有導電膜塗佈於基體上，且該導電臈具有導 電銀漿及導電被漿，絕緣層覆蓋於該基體及該導電膜上且裸露出部分導電 膜，複數個反應區覆蓋於絕緣層裸露之導電膜上，複數個反應臈層覆蓋於 反應區上，墊片用以放置反應膜層與反應區並覆蓋於絕緣層上，且使反應 鲁 Μ上方留有距離用以吸入樣品，保護顧以保護反應臈層而覆蓋於塾片 上，且该保護膜上開設有用以通氣之透孔，並於該透孔周圍設有用以方便 辨識樣品吸入口的圖案。 然而其缺點在其反應區位於頂端，檢體不易滴落其中，且不易被吸入 反應區中。 請參照第6圖所示，其為美國專利號碼第5, 997, 817號，主要揭示- 種虹吸式片狀電極試片，包括一電絕緣基板，位於其同一面上之一電極系 充電、、邑緣層覆蓋於電極系統上界定出-反應區及連接偵測器之陰陽極 接頭反應臈覆蓋於反應區電極上，一親水性薄膜覆蓋於反應膜上方， 取後再貼上-具有反應區大小透明視窗之上蓋膠顧定。然而此試片之檢 體吸入口只有_個，方向位於右側，其缺點在於··檢體施加入口只有一個 且方位也固定一個方向。 【發明内容】 、”聖ρ針對蝻述先前技術之缺點，而研究出一種可微量取樣電化學 怎 、片該5式片端緣設有一反應區，且該反應區係凸出於試片且 1269036 呈全面開放狀，因此僅需微量之檢體， 應區中。 即可以毛細擴散方式輕易被吸入反 種可微量取樣電化學生物感測器試 該等基板係由一第一基板及第二基板 之反應區，該反應區為透明狀，且該 為實現上述之目的，本案提供— 片，其设有一對具電絕緣性之基板， 組成，该弟一、二基板一端設有凸出 反應區周緣係呈全面性開放狀 而在該等第-、二基板間設有—工作電極 ❿1269036 IX. Invention description: '· 【Technical field to which the invention belongs】 • This creation is an electrochemical biosensor test piece, which is a specific electrochemical method using an oxidized or reduced substance and a sample in the sample. The electron transfer is generated by the reaction, and the test piece is used for crying, and the test piece of the sample concentration is detected, and the surrounding area of the reaction area is completely open, and the test object of the eight sample is more likely to enter the reaction area, and is not restricted by the direction. . [First Technology] Calling for the improvement of the concept of home care, fast, cheap, and no need for professional operation, home self-test products are increasing, such as blood glucose equipment, electronic ear temperature grab, electronic blood pressure monitor, etc. The disposable blood glucose test piece used is the use of the electrochemical biosensor principle to detect the blood glucose concentration in the sample. At present, there are many blood glucose test strip products on the market, and the method of applying the sample is mainly divided into drip type and quantitative type (capillary type). The test piece design is usually covered with a plastic net on the reaction area to avoid pollution. However, the demand for the sample is generally greater than 3 microliters. This is the case for the elderly or senior patients with JC to collect blood samples with a blood collection needle. It must be condensed into a drop of blood before it can be accurately and adequately dripped into the reaction zone. Inconvenient 'often causing operational errors and wasting test strips. The quantitative (capillary mode) test piece requires less than 3 microliters of sample. As long as a few samples touch the population, the sample will automatically attract the reaction zone. However, the current commercial siphon system design is not a test piece. If the size is too small, it is inconvenient and easy to pollute the population. At the same time, the suction port usually has only one and the orientation is fixed, which causes inconvenience in operation. Moreover, the ash content of the test piece is less than 3 μl. The area is reached, but the reduction of the area will result in a relatively weak signal. 1269036, which is shown in Fig. 1, which is U.S. Patent No. 5,762,770, which discloses a drip-type sheet electrode test piece comprising an electrically insulating substrate having a two-pole electrode system on the same side; An electrically insulating layer covers the electrode system and defines a reaction zone and a cathode and a cathode joint connecting the detectors; a reaction film is covered in the reaction zone, and a plastic mesh is covered over the reaction zone, and finally a reaction zone is attached The open tape fixes the plastic net. However, the sample application method of such a test piece must be dripped in the direction above the reaction zone so that the direction in which the sample is dropped in the reaction zone is naturally limited. At the same time, the sample requirement must be above 9 μL. The eye is shown in Fig. 2, which is U.S. Patent No. 5,12, (4), which mainly discloses a capillary-like electrode chip test piece comprising an electrically insulating substrate having an electrode system on the same surface, The electrically insulating layer, the reaction is difficult, and the hydrophilic upper cover plate located above the kneading surface forms a sample carrying reaction zone space, wherein the sample carrying reaction zone space has a sample suction port and a gas discharge port. The preparation is to apply a solution of a slow-acting methyl cellulose (Carb〇Xyiflethylcellul〇se, CMC) to the reaction zone of the substrate, and to form a hydrophilic polymer CMC layer after drying; The solution of Gluc〇se oxidase is coated thereon to form G_, and then the hydrophilic polymer-polyethylene-pyridylpyrene (P〇lyvinylpyrrolid〇ne) solution is applied to the third After the secondary drying, a protective film (PVP layer) is formed, in which an organic floating solution containing an electron mediator is sprayed thereon, and after drying, the reaction film is formed. v The capillary mode of the capillary electrode test piece has the disadvantage that: first, the completion of the reaction film requires four/short-domain CMC layers for improving the hydrophobicity of the electrode surface of the reaction zone; forming a (10) layer, hydrophilic 1269036 protective film PVP The layer and the electron media layer must be dried before each layer is formed, and the process steps are cumbersome. Second, after the method of applying the sample is to contact the tip of the electrode test piece, the sample is automatically introduced into the reaction zone, but only one position can apply the sample. Third, the test piece is too small to take inconvenient and easy to contaminate the sample to inhale π 〇 Please refer to Figure 3, which is the Taiwan Patent No. 573735 patent, which is a blood glucose test piece, which includes: a first substrate; a first electrode disposed on the first substrate, one end being an electrode connection point, and the other end being an electrode enzyme active region; an insulating layer covering the first substrate and acting on the electrode enzyme active region Having an opening; an enzyme film applied to the electrode enzyme active region at the opening of the insulating layer; and a second substrate covering the uppermost layer of the blood glucose test strip and having a second electrode on the bottom surface, One end of the second electrode is an electrode connection point, and the other end is an electrode enzyme action region. The insulating layer of the blood glucose test strip further forms a slit for siphoning, and an opening is formed near the end of the slit to serve as a venting hole for siphoning. Although such a structure can solve the above problems, the opening has a certain directionality, so the area is relatively reduced, and the direction and range of the drop are also limited. Furthermore, it is necessary to form an opening near the end of the slit to serve as a venting hole for siphoning, which increases the difficulty in use and manufacture. As shown in Fig. 4, it is U.S. Patent No. 6,258,229 β1, which mainly discloses a capillary test piece having a slimmer action mode of less than 丨 microliter, including an electrically insulating substrate, and a substrate on the same side. A three-pole electrode system, an insulating intermediate layer to form a 1] type channel, and a 1269036-· having a capping layer above the hole, however, the disadvantage is that the sample inlet port has only one and is located at the top end of the test piece. Please refer to FIG. 5, which is a patent of Taiwan Patent Publication No. 570192, which is a thick tantalum electrode biochemical test strip having a conductive film coated on a substrate, and the conductive crucible has a conductive silver paste and a conductive paste, an insulating layer covering the substrate and the conductive film and exposing a portion of the conductive film, a plurality of reaction regions covering the exposed conductive film of the insulating layer, and a plurality of reactive layer covering the reaction region, and the gasket is used The reaction film layer and the reaction zone are placed on the insulating layer, and a distance is left above the reaction rectification for inhaling the sample, and the protection layer covers the enamel layer to protect the reaction layer, and the protective film is opened on the film. A through hole is ventilated, and a pattern around the through hole for conveniently identifying the sample inlet is provided. However, its disadvantage is that the reaction zone is at the top end, the sample is not easily dripped therein, and is not easily inhaled into the reaction zone. Please refer to FIG. 6, which is U.S. Patent No. 5,997,817, which discloses a siphon type sheet electrode test piece comprising an electrically insulating substrate, one of which is charged on the same side of the electrode. The ruthenium layer covers the electrode system and defines a reaction zone and a cathode-anode junction reaction detector attached to the reaction zone, and a hydrophilic film covers the reaction film, and then is attached and then reacted. The size of the area is transparent above the window. However, the specimen has only _ of the sample suction ports and the direction is on the right side. The disadvantage is that the specimen has only one inlet and one orientation is fixed. SUMMARY OF THE INVENTION "Study on the shortcomings of the prior art, and to develop a micro-sampling electrochemical method, the film has a reaction zone at the end of the 5-piece piece, and the reaction zone is protruded from the test piece and 1269036 It is fully open, so only a small amount of sample is needed, which can be easily inhaled by capillary diffusion. The micro-sampling electrochemical biosensor can be used to test the substrate from a first substrate and a second substrate. In the reaction zone, the reaction zone is transparent, and in order to achieve the above object, the present invention provides a sheet, which is provided with a pair of electrically insulating substrates, and the first and second substrates are provided with a convex reaction zone at one end. The peripheral system is fully open and has a working electrode between the first and second substrates.
及一參考電極’該等電極間保持適當距離，且該等電極-端並延伸至反應 區中如此在進行;^測時’可因反應區周緣係呈全面性開放狀，使檢體 易於以毛細概枝狀反舰，衫開口綠_，綠取樣，令檢體 需求量小於3微升’可減少病患擠血之龄，且缝需健欧，以降低 母次操作檢體量不同而造成之測量誤差。 本創作之再_目的，在提供_種可微量取樣電化學生物感測器試片， 其反應區設有一親水性反應膜，該親水性反應膜覆蓋於工作電極及參考電 極，該反應膜用以與檢體接觸而產生專一性電化學反應，且該反應臈係由 親水性高分子及酵素構成，以使檢體容易進入反應區。 本創作另一目的，在提供一種可微量取樣電化學生物感測器試片，其 反應區邊緣係呈凸出結構，不僅觸覺易於辨識且能使檢體易於吸入，以減 少檢體進入反應區產生停頓之現象((jose hes i tat i on)。 【實施方式】 本新型揭示一種可微量取樣電化學生物感測器試片，利用具有氧化或 還原物質與檢體發生專一性生化反應而產生電子轉移之電化學生物感測器 試片，其配合檢測儀器使用，可專一且靈敏地量測檢體之濃度含量。 1269036 : 請參照第7、8圖所示，該試片上設有相對之第-、二基板u、12 一 ' 一、二基板11、12係呈平板狀且具電絕緣性，且該第一 u 一 I板 11、12 — 周緣除了兩端外之相對應面具有黏性膠(其爲本創作之一 貝她方式，惟熟 悉該項技術者，可以其他方式取代），以使第—、二基板u id騰合在一 起，而無覆膠部分具有親水性，其中第二基板12 一端未能完全覆蓋在第一 基板11上，而該第一、二基板u、12另一端則設有一向外凸出之反應區 13，該反應區13為透明，且該反應區13周緣係呈凸出狀，又該反應區η • 周緣並呈全面性開放狀，不僅易於辨識且能使檢體易於以毛細擴散方式吸 入，以減少檢體進入反應區產生停頓之現象(如第8圖所示）。 另，該等基板11、12間夾置有一工作電極21及一參考電極22，嗲工 作電極21及參考電極22並保持一定間距，且該工作電極21及一參考電極 22 —端深入反應區13，另一端則凸出於第二基板12之另一端，且較第一 基板11短’而載於第一基板11該端上。 再者，該反應區13上覆蓋有親水性反應膜30，該等親水性反應膜3〇 ® 並將工作電極21及參考電極22覆蓋其下，於本實施例為親水性酵素，係 由氧化或還原物質、親水性高份子、電子媒介物、界面活性劑及緩衝溶液 之配方所組成，用以與檢體接觸而產生生化反應。 使用時，請參照第9、10圖所示，如此，在進行檢測時，可因反應區 13係凸出於基板外，且反應區13周緣係呈全面性開放狀，使檢體易於以毛 細擴散方式吸入反應區13内，不受方位限制，方便取樣，更因檢體藉由覩 水性反應膜30擴散導入反應區13中進行生化反應，檢體需求量更是小於3 1269036 微升，可減少病患擠血之痛苦，且檢體需求量固定，以降低每次操作檢體 v 量不同而造成之測量誤差。 .综上所述’本創作在物品、形狀、構造、結構上屬首先創作，且可改 良習用技術之各種缺點，在使用上能增進功效，合於實用，充分#人新， 專利要件’實為一理想之創作。 〃 【圖式簡單說明】 第1圖為美國專利第5, 762, 77Q號專利案之示意圖。 • 第2圖為美國專利第5,12M2G號專利案之示意圖。 第3圖為台灣專利第573735號專利案之示意圖。 第4圖為美國專利第6, 258, 229β1號專利案之示意圖。 第5圖為台灣專利第57〇192號專利案之示意圖。 第6圖為美國專利第5, 997, 817號專利案之示意圖。 第7圖為本創作之分解示意圖。 第8圖為本創作之組合示意圖。And a reference electrode 'the electrodes are maintained at an appropriate distance, and the electrode-ends are extended into the reaction zone so as to be performed; when the test is performed, the periphery of the reaction zone is comprehensively open, so that the sample is easy to The capillary-like anti-ship, the green opening of the shirt, the green sampling, so that the demand for the sample is less than 3 microliters, can reduce the age of the patient's blood, and the seam needs to be healthy, so as to reduce the amount of the mother's operation. The measurement error caused. The purpose of the present invention is to provide a micro-sampling electrochemical biosensor test piece having a hydrophilic reaction film covering the working electrode and the reference electrode, and the reaction film is used for the reaction film. A specific electrochemical reaction is generated by contact with the sample, and the reaction oxime is composed of a hydrophilic polymer and an enzyme to facilitate entry of the sample into the reaction zone. Another object of the present invention is to provide a micro-sampling electrochemical biosensor test piece having a convex structure at the edge of the reaction zone, which is not only easy to recognize and can easily inhale the sample, thereby reducing the entry of the sample into the reaction zone. The phenomenon of occurrence of pause ((jose hes i tat i on). [Embodiment] The present invention discloses a micro-sampling electrochemical biosensor test piece, which is produced by using a specific biochemical reaction with an oxidized or reduced substance and a sample. The electronically transferred electrochemical biosensor test piece, which is used in conjunction with the detection instrument, can measure the concentration content of the sample in a specific and sensitive manner. 1269036 : Please refer to Figures 7 and 8, the test piece is provided with a relative The first and second substrates u, 12 and the first and second substrates 11 and 12 are flat and electrically insulating, and the first u-I plates 11 and 12 have a sticky surface on the periphery except for the two ends. Sexual glue (which is one of the creative methods of the creation, but those who are familiar with the technology, can be replaced by other means), so that the first and second substrate u id are together, and the non-coated part is hydrophilic, wherein Second substrate 12 The end portion is not completely covered on the first substrate 11, and the other ends of the first and second substrates u, 12 are provided with an outwardly convex reaction region 13, the reaction region 13 is transparent, and the reaction region 13 is peripherally It has a convex shape, and the reaction zone η • the periphery is open and comprehensive, which is not only easy to identify but also makes the sample easy to be inhaled by capillary diffusion, so as to reduce the pause of the sample entering the reaction zone (Fig. 8) In addition, a working electrode 21 and a reference electrode 22 are interposed between the substrates 11 and 12, and the working electrode 21 and the reference electrode 22 are kept at a certain distance, and the working electrode 21 and a reference electrode 22 are end-to-end. The reaction zone 13 is covered with hydrophilicity. The other end is protruded from the other end of the second substrate 12 and is shorter than the first substrate 11 and is carried on the end of the first substrate 11. Further, the reaction zone 13 is covered with hydrophilicity. The reaction film 30, the hydrophilic reaction film 3〇® and the working electrode 21 and the reference electrode 22 are covered thereunder. In this embodiment, the hydrophilic enzyme is an oxidized or reduced substance, a hydrophilic high molecular weight, and an electron vehicle. , surfactant and buffer solution formula The composition is used for contact with the sample to generate a biochemical reaction. When using, please refer to the figures 9 and 10, so that when the detection is performed, the reaction zone 13 may protrude from the substrate and the periphery of the reaction zone 13 The invention has a comprehensive open shape, so that the sample is easily inhaled into the reaction zone 13 by capillary diffusion, and is not restricted by orientation, and is convenient for sampling, and the sample is diffused into the reaction zone 13 for biochemical reaction by the hydrophobic reaction membrane 30, The sample requirement is less than 3 1269036 microliters, which can reduce the pain of patients' blood squeezing, and the sample needs are fixed, so as to reduce the measurement error caused by the difference of the amount of sample v per operation. This creation is the first creation in the article, shape, structure and structure, and can improve various shortcomings of the conventional technology. It can enhance the effect in use and be practical, fully #人新, patent element 'is an ideal creation. 〃 [Simple description of the drawing] Fig. 1 is a schematic diagram of the patent of U.S. Patent No. 5,762, 77Q. • Figure 2 is a schematic diagram of U.S. Patent No. 5,12M2G. Figure 3 is a schematic diagram of Taiwan Patent No. 573735. Figure 4 is a schematic illustration of U.S. Patent No. 6,258,229, filed No. 1, patent. Figure 5 is a schematic diagram of Patent No. 57〇192 of Taiwan Patent. Figure 6 is a schematic illustration of U.S. Patent No. 5,997,817. Figure 7 is an exploded view of the creation. Figure 8 is a schematic diagram of the combination of the creations.
第9圖為本創作組合之平面透視圖。 第10圖為本創作實施時之斷面示意圖 【主要元件符號說明】 第一基板 11 第二基板 12 反應區 13 工作電極 21 參考電極 22 11 1269036 親水性反應膜 30Figure 9 is a plan perspective view of the creative combination. Figure 10 is a schematic cross-sectional view of the present invention. [Main component symbol description] First substrate 11 Second substrate 12 Reaction zone 13 Working electrode 21 Reference electrode 22 11 1269036 Hydrophilic reaction film 30
⑧ 128 12