201128186 六、發明說明： 【發明所屬之技術 [醒]本發明有關一種生物檢測元件’尤指一種利用表面聲波 (surface acoustic wave,簡稱SAW)檢測浪態檢艘之 生物檢測元件；本發明另有關於上述之生物檢測方法 特別關於一種透過一感測聚合物重量減少而分析表面聲 波的生物檢測方法。 【先前技術】 0 [0002] 一般檢驗生物性樣品或檢體的撿測方式主要有二，〆種 是將前述之樣品或檢鱧進行萃取、濃縮、沉澱或利用質 譜、層析等分析、分離步驟，純化而進行檢驗；另一種 方式則利用特異(專一)性反應直接量測複雜樣品中的待 測物，例如：利用生物晶片或感測器量（感）測待測物的 存在與否。 闺乡數生物感測器係由-辨識物質、一換能器以及一訊號 處理器所組成’當待測物與辨識物質發生反應時，換能 〇 11可狀應產生的_献學變化㈣成電氣訊號，並 經由訊號處理器分析而獲致感測結果。由上可知，生物 感測器之辨識物質必須對應待測物產生特異性，以準確 備測待測物質，常見的辨識物質例如：生物組織、微生 物、胞器、細胞受體、酵素、抗原抗體等。 _ 「圖1」係-種表面聲波元件（SAW device)1，包含一壓 電基板11、分別形成於壓電基板u表面兩端之第一換能 器12與第-換成器13以及一外部電路（圖中未示）。該第 -、二換能器12，13包含複數個指叉式電極14，當外部 099103739 表單編號A0101 第3頁/共18頁 0992007016-0 201128186 電路施予第一換能器12一電氣訊號時，該第一換能器12 可透過壓電效應將電氣訊號轉換為表面聲波，並沿著壓 電基板U表面傳播，而後再經由第二換能器13之指叉式 電極14，將表面聲波轉為電氣訊號而自外部電路輸出。 由於表面聲波的傳遞極易受到外部環境的影響，因此非 常適合作為感測元件。進一步地，由於表面聲波元件體 積小、重量輕、成本低、可以半導體大量批次製造的優 點，且其靈敏度高，具備高訊號/雜訊比，因此近年來衍 生用作於生物感測器。 [0005] [0006] 其中’中華民國專利公開第制08048Θ7號之「化學或生 物物質感應器」係將指標器.生.物分子混:合於溶膠—凝膠 衍生物中’藉以針對氣體中的待測物進行檢測；美國專 利第 5658732 號之「Assay method for biological target complexes on the surface of a bio- sensor」以及第 5814525 號之「Piezoelectric bi〇_ .:: ,·,：： sensor with a ladder polymer substrate coat-ing」，分別揭露一種檢驗生物檢體的方法及壓電生物感 測器’係利用特異的辨識物質捕捉待檢測之目標物，該 些辨識物質包含具活性的耦合位置（active binding sites)，藉以與待測檢體（analyte)形成鍵結、偶合而 偵測待測物。 相較於上述習知技術，本發明提供一種生物檢測元件， 結合一表面聲波元件與一感測聚合物，利用待測物與感 測聚合物反應而造成感測聚合物質量實質上減少，藉以 偵測待測物的存在。 099103739 表單編號A0101 第4頁/共18頁 0992007016-0 201128186 【發明内容】 [0007] 承上，本發明之目的在於提供一種生物檢測元件及其檢 ' 測方法，用以快速、靈敏地檢測液態檢體中欲檢測之待 測物。 [0008] 為了達成前述目的，本發明提供一種生物檢測元件，其 結合一表面聲波元件與一感測聚合物，包含：一壓電基 板、一第一換能器、一第二換能器以及一感測聚合物。 該感測聚合物形成於該壓電基板上，能與欲檢測之待測 物發生反應；該第一換能器與第二換能器形成該壓電基 Ο 板上，位於該感測聚合物相對應的兩端，並分別定義為 表面聲波之輸出端與輸入端。其中，該生物檢測方法係 透過感測聚合物與該待測物發生反應，致使表面聲波因 感測聚合物的重量實質上減少而改變其特性，藉由偵測 、分析表面聲波的特性改變，以檢測、判別液態檢體中 待測物的屬性與含量。有關本發明的詳細技術内容及較 佳實施例，配合圖式說明如後》 ^ 【實施方式】 [0009] 為了方便理解本發明的技術所在，以下的說明係整合生 物檢測元件及其檢測方法一同詳細陳述。有關本發明之 詳細說明及技術内容，現配合圖式說明如下：首先，「 圖2」顯示本發明之生物檢測元件2—實施例的外觀立體 示意圖，該生物檢測元件2結合一表面聲波元件20與一感 測聚合物21，藉以檢測液態檢體中之待測物；該生物檢 測元件2包含：一壓電基板201、一第一換能器202、一第 二換能器203、一感測聚合物21以及一外部電路（圖中未 099103739 表單編號A0101 第5頁/共18頁 0992007016-0 201128186 示）。該感測聚合物21形成於壓電基板2〇1上一側，可與 特異（對應）之待測物產生反應。對於「特異」一詞，以 下將有說明。該第一換能器202與該第二換能器2〇3形成 於該壓電基板201同側，位於該感測聚合物21的對應兩端 ，並分別與外部電路電性連接，且各自包含複數個指又 式電極204 ;第一換能器202可自外部電路接收一電氣訊 说，並將a亥電氣说號轉換而產生表面聲波；該表面聲波 傳經反應後之感測聚合物21而改變特性，並由第二換能 器203接收而轉換為一電氣訊號，分析、比對該電氣訊號 便可判別該待測物的存在與否，或對待測物進行定量 、定性分析。 [0010] [0011] 在上述之實施例中，該表面聲波灸#2〇係為一水平剪力 表面聲波元件（shear horizontal SAW，簡稱SH-SAW) ’ β玄壓電基板2〇1的材質例可為石英（Quartz)、銳酸鐘 (LiNb〇3)及鈕酸鐘（LiTaQ3)。 ：* .： 請再配合參閱「圖3-1」至「圖3-3」所示，其為本發明 之生物檢測方法之步驟示意圖。丨首先，針對欲檢測之待 測物4(顯示於「圖3-2」）備製一相對應的生物檢測元件 2(如「圖3-1」所示），即該感測聚合物21係對應欲檢測 之待測物4 ’上述「對應」係指該感測聚合物21可檢測出 該待測物4 ;接著，取用適量之液態檢體3與該感測聚合 物21反應（如「圖3-2」所示）。在一實施例中，該生物檢 測元件2於該感測聚合物21上可設置一容置空間2〇5 ;該 容置空間205可容置該液態檢體3，使液態檢體3中之待測 物4與感測聚合物21產生反應，而後再移（清、洗）除液態 099103739 表單編號A0101 第6頁/共18頁 0992007016-0 201128186 檢體3。由於待測物4與感測聚合物21反應後，會造成該 感測聚合物21的重量實質上減少（如「圖3-3」所示）’甚 至改變液態檢體3之導電、黏滯度，如此會導致傳導經過 感測聚合物21之表面聲波發生中心頻率、相位變化，或 是表面聲波的能量損失，因此可透過分析表面聲波之變 化模式，判讀液態檢體3中是否含有欲檢測之待測物4， 並藉以對該待測物4進行定量或定性分析。 [0012] Ο ❹ [0013] 須說明的是，前述感測聚合物21產生重量上的實質減少 ’係指該感測聚合物21的體積、面積、厚度等物理尺寸 ，由於與待測物4反應而減少（與質量、密度減少同義）， 致使傳遞過之表面聲波改變其物理特性。因此，該感測 聚合物21須與待測物4產生特異性（sPecificity) ’所 謂的特異性係指感測聚合物21僅與待測物4發生反應，而 不和液態檢體3中其他的物質反應。另一方面，在另—實 施例中，係以水、生理食鹽水或I不與該感巧1丨聚合物21反 應之緩衝液清洗該感測聚合物21，以移除該液態檢體3， 同時可伴隨以烘乾或吹乾等不破壞該感測聚合物21的方 式去除濕氣；在另一實施例>，該感測聚合物21可為一 固態物質。 透過上述之生物檢測元件及方法，可用以快速檢測待測 物4之屬性及含量，而有助於醫療疾病的判定或貢獻於生 醫研究。與先前技術相較，前述之相關前案係於薄膜上 形成捕捉分子或物質，以抓取性質特異之待測物，例如 抗原與抗體的組合；而本發明異於前述先前技術之處， 在於本發明之感側聚合物21本身與液態檢體3中之待測物 099103739 表單編號A0101 第7頁/共18頁 0992007016-0 201128186 4發生反應，並藉由偵測表面聲波對應該感測聚合物21之 物理量值減少發生的改變，來檢測待測物4。 [0014] 在一實施例中，本發明可應用於酵素（enzyme)的檢測， 茲具體說明如下：為了檢測酵素，於本實施例中，該感 測聚合物21係可與酵素反應之一明膠（gelatin)或水膠 (hydrogel)，固定塗佈於該壓電基板201。明膠或水膠 在常溫或常溫之下為固態物質，當其與酵素（例如蛋白質 水解酵素（proteolytic enzyme)、明膠酵素）發生水解 反應後，部分的明膠或水膠會分解為氨基酸，引起液化 (liquefaction)現象而水解成液態，並隨著移（清、洗 )除液態檢體3的過程而被帶離。因此，此特異性質可應 用於含有酵素的液態檢體3檢測。 [0015] 以膀胱癌或泌尿相關疾病的檢測為例（液態檢體3為尿液） ，正常的尿液（液態檢體3)幾無或僅含微量酵素，以本發 明之生物檢測元件進行檢測時，表面聲波的特性改變不 大；然而，當膀胱出現癌細胞或罹患相關疾病時，酵素（ 待測物4)量會大增，而導致反應後的明膠（感測聚合物 21)物理量（質量）明顯減少，而可偵測上述疾病。「圖4 」顯示本發明之生物檢測元件2檢測酵素之實驗結果，其 中實驗組以含有酵素之水溶液模擬液態檢體3，對照組則 以純水模擬液態檢體3。由實驗結果可知，在兩種液態檢 體3與感測聚合物21反應的1 800秒内，其表面聲波之頻率 相異極大，而可明顯區分何者是帶有酵素之液態檢體3。 感測聚合物21減少的量值與表面聲波改變呈現出的特性 有關，因此可用以進行酵素的定量分析，以達到疾病的 099103739 表單編號A0101 第8頁/共18頁 0992007016-0 201128186 分析與判定。 [0016] 惟以上所述者，僅為本發明之較佳實施例，非欲侷限本 發明專利之專利保護範圍，故舉凡運用本發明說明書及 圖式内容所為之等效變化與修飾，均同理包含於本發明 之權利保護範圍，合予陳明。 【圖式簡單說明】 [0017] 本發明的實施方式係結合圖式予以描述： [0018] 「圖1」為一習知表面聲波元件之外觀立體示意圖； [0019] 「圖2」為本發明之生物檢測元件一實施例之外觀立體示 意圖； [0020] 「圖3_1」至「圖3_3」為本發明之生物檢測方法之步驟 不意圖，以及 [0021] 「圖4」為本發明之生物檢測元件檢測酵素之實驗結果圖 【主要元件符號說明】 [0022] 1， 20 · · •·..表面聲波元件 [0023] 2 · · · · • ••生物檢測元件 [0024] 3 · ·—— •··液態檢體 [0025] 4 ———— ...待測物 [0026] 11, 201 · ...· ·壓電基板 [0027] 12, 202 · .* .·第一換能器 [0028] 13, 203 · ....第二換能器 表單編號A0101 第9頁/共18頁 〇 099103739 0992007016-0 201128186 [0029] 1 4, 204 · · [0030] 21.....201128186 VI. Description of the invention: [Technology according to the invention [Wake] The present invention relates to a biological detecting element', in particular to a biological detecting element for detecting a wave state using a surface acoustic wave (SAW); The above-described bioassay method relates in particular to a bioassay method for analyzing surface acoustic waves by sensing a decrease in weight of a polymer. [Prior Art] 0 [0002] There are two main methods for detecting biological samples or samples. The above samples are extracted, concentrated, precipitated or analyzed by mass spectrometry, chromatography, etc. The step is to carry out the test by purification; the other method uses a specific (specific) reaction to directly measure the analyte in the complex sample, for example, using a biochip or a sensor quantity (sensing) to measure the presence or absence of the analyte. The Xiangxiang number biosensor consists of - identification material, a transducer and a signal processor. When the analyte and the identification substance react, the transducer 〇11 can produce a change in the state (4). It is an electrical signal and is analyzed by a signal processor to obtain a sensing result. It can be seen from the above that the identification substance of the biosensor must be specific to the object to be tested, so as to accurately prepare the substance to be tested. Common identification substances such as biological tissues, microorganisms, organelles, cell receptors, enzymes, antigen antibodies Wait. _ "FIG. 1" is a surface acoustic wave device (SAW device) 1 including a piezoelectric substrate 11, first transducers 12 and first-converters 13 and one formed at both ends of the surface of the piezoelectric substrate u, respectively External circuit (not shown). The first and second transducers 12, 13 include a plurality of interdigitated electrodes 14 when the external 099103739 form number A0101 page 3 / 18 pages 0992007016-0 201128186 circuit is applied to the first transducer 12 an electrical signal The first transducer 12 converts the electrical signal into a surface acoustic wave through a piezoelectric effect, and propagates along the surface of the piezoelectric substrate U, and then transmits the surface acoustic wave via the interdigitated electrode 14 of the second transducer 13. It is converted to an electrical signal and output from an external circuit. Since the transmission of surface acoustic waves is highly susceptible to the external environment, it is very suitable as a sensing element. Further, since the surface acoustic wave element has a small volume, a light weight, a low cost, and can be mass-produced in a large number of semiconductors, and has high sensitivity and high signal/noise ratio, it has been used as a biosensor in recent years. [0006] [0006] The "Chemical or Biological Matter Sensor" of the Republic of China Patent Publication No. 08048Θ7 is a product of the indicator, the material molecule, and the sol-gel derivative. The test object is tested; "Assay method for biological target complexes on the surface of a bio-sensor" and "Piezoelectric bi〇_.::,·::: sensor with a", US Pat. No. 5,587,732 Ladder polymer substrate coat-ing", respectively, discloses a method for testing a biological sample and a piezoelectric biosensor for capturing a target to be detected by using a specific identification substance, which contains an active coupling position (active The binding sites are used to form a bond and couple with the analyte to detect the object to be tested. Compared with the above-mentioned prior art, the present invention provides a biodetection element that combines a surface acoustic wave component with a sensing polymer to cause a substantial decrease in the mass of the sensing polymer by reacting the analyte with the sensing polymer. Detect the presence of the analyte. 099103739 Form No. A0101 Page 4 / 18 pages 0992007016-0 201128186 SUMMARY OF THE INVENTION [0007] In view of the above, it is an object of the present invention to provide a biodetection element and a method for detecting the same, for quickly and sensitively detecting a liquid state The object to be tested in the specimen. [0008] In order to achieve the foregoing object, the present invention provides a bio-detection element that combines a surface acoustic wave component and a sensing polymer, including: a piezoelectric substrate, a first transducer, a second transducer, and A sensory polymer. The sensing polymer is formed on the piezoelectric substrate and can react with the object to be detected; the first transducer and the second transducer form the piezoelectric substrate, and the sensing polymerization is located The corresponding ends of the object are defined as the output end and the input end of the surface acoustic wave. Wherein, the biological detection method reacts the sensing polymer with the object to be tested, so that the surface acoustic wave changes its characteristics by substantially reducing the weight of the sensing polymer, and by detecting and analyzing the characteristic change of the surface acoustic wave, To detect and discriminate the properties and content of the analyte in the liquid sample. The detailed technical content and preferred embodiments of the present invention are described in conjunction with the drawings. [Embodiment] [0009] In order to facilitate the understanding of the technology of the present invention, the following description is integrated with the biological detecting component and its detecting method. Detailed statement. The detailed description and technical contents of the present invention will now be described with reference to the following drawings: First, "FIG. 2" shows an external perspective view of a biodetection element 2 - an embodiment of the present invention, which incorporates a surface acoustic wave element 20 And sensing the polymer 21 to detect the object to be tested in the liquid sample; the biological detecting element 2 comprises: a piezoelectric substrate 201, a first transducer 202, a second transducer 203, and a sense The polymer 21 and an external circuit are measured (not shown in the figure 099103739, Form No. A0101, page 5/18 pages, 0992007016-0, 201128186). The sensing polymer 21 is formed on the side of the piezoelectric substrate 2〇1 and reacts with a specific (corresponding) analyte. For the word "exclusive", the following will be explained. The first transducer 202 and the second transducer 2〇3 are formed on the same side of the piezoelectric substrate 201, and are located at corresponding ends of the sensing polymer 21, and are electrically connected to external circuits, respectively. The plurality of finger-receiving electrodes 204 are included; the first transducer 202 can receive an electrical signal from an external circuit and convert the a-hai electrical number to generate a surface acoustic wave; the surface acoustic wave passes through the reacted sensing polymer 21 The characteristic is changed and received by the second transducer 203 to be converted into an electrical signal, and the electrical signal can be analyzed and compared to determine whether the object is present or not, or the object to be measured is quantitatively and qualitatively analyzed. [0011] In the above embodiment, the surface acoustic wave moxibustion #2 is a material of a horizontal shear surface acoustic wave component (shear horizontal SAW, referred to as SH-SAW) 'β-shaped piezoelectric substrate 2〇1 Examples may be quartz (Quartz), sharp acid clock (LiNb〇3), and button acid clock (LiTaQ3). :* .: Please refer to the "Figure 3-1" to "Figure 3-3" for further description of the steps of the biological detection method of the present invention.丨 First, prepare a corresponding bio-detection element 2 (shown in Figure 3-1) for the object to be tested 4 (shown in Figure 3-2), that is, the sensing polymer 21 Corresponding to the test object 4 to be detected, the above "corresponding" means that the sensing polymer 21 can detect the test object 4; and then, an appropriate amount of the liquid sample 3 is reacted with the sensing polymer 21 ( As shown in Figure 3-2. In an embodiment, the bio-detecting element 2 can be provided with an accommodating space 2〇5 on the sensing polymer 21; the accommodating space 205 can accommodate the liquid sample 3, so that the liquid sample 3 can be The analyte 4 reacts with the sensing polymer 21, and then moves (clear, wash) and removes the liquid 099103739 Form No. A0101 Page 6 / 18 pages 0992007016-0 201128186 Sample 3. Since the analyte 4 reacts with the sensing polymer 21, the weight of the sensing polymer 21 is substantially reduced (as shown in "Fig. 3-3"), and even the conductivity and viscosity of the liquid sample 3 are changed. Degree, which causes the center frequency, phase change, or energy loss of the surface acoustic wave to be transmitted through the surface acoustic wave of the sensing polymer 21, so that the liquid sample 3 can be read to determine whether or not the liquid sample 3 is to be detected by analyzing the change pattern of the surface acoustic wave. The object to be tested 4 is used for quantitative or qualitative analysis of the object to be tested 4. [0012] [0013] It should be noted that the above-described sensing polymer 21 produces a substantial decrease in weight', which refers to the physical size of the volume, area, thickness, etc. of the sensing polymer 21, due to the object to be tested 4 The reaction is reduced (synonymous with mass and density reduction), causing the transmitted surface acoustic waves to change their physical properties. Therefore, the sensing polymer 21 must be sPecificity with the analyte 4, the so-called specificity means that the sensing polymer 21 reacts only with the analyte 4, and not with the liquid sample 3 Material reaction. On the other hand, in another embodiment, the sensing polymer 21 is washed with water, physiological saline or a buffer which does not react with the sensor 1 to remove the liquid sample 3 At the same time, moisture may be removed in a manner that does not destroy the sensing polymer 21 by drying or drying; in another embodiment, the sensing polymer 21 may be a solid material. The biodetection element and method described above can be used to quickly detect the property and content of the test object 4, and contribute to the judgment of medical diseases or contribute to biomedical research. In contrast to the prior art, the foregoing related pre-forms form a capture molecule or substance on the film to capture a property-specific analyte, such as a combination of an antigen and an antibody; and the present invention is different from the foregoing prior art in that The sensory side polymer 21 of the present invention reacts with the object to be tested in the liquid sample 3, 099103739, Form No. A0101, page 7 / page 18, 0992007016-0, 201128186 4, and detects the polymerization by detecting surface acoustic waves. The physical quantity of the object 21 is reduced by a change to detect the object 4 to be tested. [0014] In one embodiment, the present invention is applicable to the detection of an enzyme, which is specifically described as follows: In order to detect an enzyme, in the present embodiment, the sensing polymer 21 is one of gelatin which can react with an enzyme. (gelatin) or hydrogel is fixedly applied to the piezoelectric substrate 201. Gelatin or water gel is a solid substance at normal temperature or normal temperature. When it is hydrolyzed with an enzyme (such as proteolytic enzyme or gelatin), part of the gelatin or water gel will be decomposed into amino acids, causing liquefaction ( The liquefaction phenomenon is hydrolyzed into a liquid state and is carried away as the liquid sample 3 is removed (cleared, washed). Therefore, this specific substance can be applied to the detection of a liquid sample 3 containing an enzyme. [0015] Taking the detection of bladder cancer or urinary related diseases as an example (liquid sample 3 is urine), normal urine (liquid sample 3) contains little or only trace amounts of enzyme, and is carried out by the biodetection element of the present invention. At the time of detection, the characteristics of surface acoustic waves do not change much; however, when cancer cells or related diseases occur in the bladder, the amount of enzyme (subject 4) increases greatly, resulting in the physical quantity of gelatin (sensing polymer 21) after the reaction. (Quality) is significantly reduced, and the above diseases can be detected. Fig. 4 shows the results of an experiment for detecting an enzyme by the biosensor 2 of the present invention, wherein the experimental group simulates a liquid sample 3 with an aqueous solution containing an enzyme, and the control group simulates a liquid sample 3 with pure water. It can be seen from the experimental results that the frequency of the surface acoustic waves is extremely different within 1 800 seconds of the reaction between the two liquid samples 3 and the sensing polymer 21, and it is possible to clearly distinguish which is the liquid sample with the enzyme 3 . The magnitude of the decrease in the sensing polymer 21 is related to the characteristics exhibited by the surface acoustic wave change, and thus can be used for quantitative analysis of the enzyme to achieve the disease 099103739 Form No. A0101 Page 8 of 18 0992007016-0 201128186 Analysis and determination . [0016] The above is only the preferred embodiment of the present invention, and is not intended to limit the scope of the patent protection of the present invention, so the equivalent changes and modifications of the present invention and the contents of the drawings are the same. It is included in the scope of protection of the present invention and is combined with Chen Ming. BRIEF DESCRIPTION OF THE DRAWINGS [0017] Embodiments of the present invention are described in conjunction with the drawings: [0018] FIG. 1 is a perspective view showing the appearance of a conventional surface acoustic wave element; [0019] "FIG. 2" is the present invention. [0020] "Fig. 3_1" to "Fig. 3_3" are the steps of the biological detection method of the present invention, and [0021] "Fig. 4" is the biological detection of the present invention. Experimental results of component detection enzymes [Description of main component symbols] [0022] 1, 20 · · • ·. Surface acoustic wave components [0023] 2 · · · · • • Biometric components [0024] 3 · · •···Liquid specimen [0025] 4 ————...Substance [0026] 11, 201 · ...··Piezoelectric substrate [0027] 12, 202 · .* .·First transduction [0028] 13, 203 · .... second transducer form number A0101 page 9 / total 18 pages 〇 099103739 0992007016-0 201128186 [0029] 1 4, 204 · [0030] 21.... .
[0031] 205 . . . ··.指叉式電極 ••感測聚合物 •容置空間 099103739 表單編號A0101[0031] 205 . . . ·.. Fork-type electrode •• Sense polymer • accommodating space 099103739 Form No. A0101
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