M316891 八、新型說明： 【新型所屬之技術領域】 •本賴賊擴增法及免疫法之檢猶置有關，更詳而言之，是—種配合核 •酸及免疫法的快速檢測裝置。 【先前技術】 核酸增幅技術及免疫法已成為生物學領域最重要的工具之一，在臨床醫學 應用領域中，舉凡感染病原、遺傳疾病...等的檢測，扮演著舉足輕重的角色。 _ -轉财產品檢測疫病為例，介紹習知糊聚合酵素連航應(p⑻結合生 物晶片的檢測流程。首先利用PCR增幅特定的核酸片段，然後將pcR增幅後的產 物加上雜合反應液後注入-晶片盒中，放到雜合反應箱中，在特定的環境溫度 下’進行雜合至少-小時(_需不斷搖晃以增強雜合反應），雜合完成後將晶 片内液體倒掉，加入清洗溶液，倒掉清洗溶液並加入特定濃度的抗生物素驗性 磷酸酵素(SA-AP)，於室溫下反應十分鐘，再將晶片内液體倒掉，加入清洗溶 液，倒掉清洗溶液並加入特定濃度的呈色劑⑽T/BCIp)，於室溫下反應十分鐘， 籲.再將晶片喊體娜，最後以水清洗，觀察晶片是否出現呈色反細判斷樣品 是否染有某種疫病。 上述檢測方法’不論是試舰擇、用量、流程、操作方式、環境控制..等 均疋以-種「知識的」、「理論化的」型態被專門人員所知悉，而所使用的試 劑及硬體也是以實驗設備的樣態呈現，並存放於實驗室中，這使得檢測方式變 得非常的複齡具有相當的困難度，以至於必需由專門人員進行，且檢測環境 被限制於實驗室中。這些原因’使得檢測工作無法交由養輯者自行實施，對 於疫病的即時防治恐造成延遲的問題。除此之外，若專門人員未完全依照標準 5 M316891 則可能使得檢測結果發生嚴重的誤 化流程進行檢測程序，或者檢驗程序錯誤 差，但卻不易被覺察。 【新型内容】 將上述繁複而具有困難度的 本案之《目的係在提出_種快速檢測裝置 檢測方法轉換為簡單而具體。 -種核酸綱法及m之快触 、x置，主要包括：一偵測板，内部具 有至少一可呈現特定檢測結果的基暂，八 、以土貝可為塑膠片、玻璃、尼龍臈、硝 化纖維膜、聚偏氣乙稀膜等其中一插·#& 種，该偵測板的一側面開設至少一鏤空的視 窗，樣品可透過該視窗而接觸該基f環蓋，結合於該偵測板的綱；若干 存放於管瓶_ ，每-管瓶可供上述_板容置其中，且辭瓶之瓶口可 與該環蓋相互固定。 與先前技術所述之檢測方法相較，本案將檢測所需之試劑、用量、流程、 操作方式等製做成序列的若干管瓶，而檢測樣品則以該偵測板為載具，且該價 測板另可呈現檢驗絲以及具有躺制成功或失_。本案使檢測方法 __變得簡單、快速、具體、且正確率提高，因此可以交由養殖業者於養殖場中自 行實施，對於疫病的檢測及防治具有即時而有效的意義。 【實施方式】 如第一、二、三圖，本案核酸擴增法及免疫法之快速檢測裝置，主要包括： 一偵測板10，其以長寬比例定義出一種窄長的板片形狀，該偵測板10内部 具有至少一可呈現特定檢測結果的基質50，該基質50可為塑膠片、玻璃、尼龍 膜、硝化纖維膜、聚偏氟乙烯膜…等。該偵測板10開設至少一鏤空的視窗11， 樣品可透過該視窗而接觸該基質50 ; M316891 - 一環蓋20，結合於該偵測板ι〇的頂端； 若干存放於管瓶30内的試劑31，每一管瓶30可供上述偵測板1〇容置其中， • 且該管瓶3〇之瓶口 32可與該環蓋20相互固定。 在本案貝知例中，上述偵測板1〇係由一面板12以及一底板a相對結合所構 成。該面板12包括了一位於頂端的柱狀結合部14、位於背面的基質定位槽巧、 以及鏤空的第一視窗ni以及第二視窗112。該基質定位槽15中設置一具特定檢 測功此的基質50。该底板13結合於該面板12的背面而完全的封閉上述的基質定 籲位槽15，並且將該基質50固定。該底板13與該面板12的結合方式係在兩者的相 對面對應位置設若干可彼此緊制嵌合的凸柱16及凹槽17。 如第、二、五、六圖，在本案實施例中，上述環蓋2〇設一可供該偵測板 10之柱狀結合部14穿合的減觀，據此職2G結合於該_板1〇的頂 端。該柩接槽21以及該柱狀結合部14之結合部面設有可相互卡合的凸環24及環 槽141 ’以便使該裱蓋2〇能與偵測板1〇能穩固地結合。該環蓋2〇與該管瓶之瓶 口32設有可相互螺合的螺紋23、33，當該横測板1〇置入該管瓶3〇中，該環蓋2〇 籲可螺合於該瓶π32。該環蓋20的頂面設一板片狀的持取部23，其板片形狀便於 手指持取，更便於在其上標註樣品的記號。 如第四圖，在本案實施例中，上述管瓶30是以一密封蓋34將内部的試劑密 封之’試劑若為性質穩以保存之化學製劑，則可為液態儲存於該管瓶中；試 劑若不易保存之活性酵素或類似品，雜冷魏燥製献末*保存於該管 瓶之瓶底，於使用時添加預定量之水液使之活化。上述的管瓶3〇依照檢測之順 序編碼，以便檢測者按順序取用。 以養殖水產品是否感染白點病毒為例，說明如何使用本案之快速檢測襄置。 M316891 - 步驟―’錄樣指示取鎌狀水絲品錢行核_歡應，待反應完 成； ' #驟二，取出本㈣着，關花棒沾取步驟—之樣品溶液，將棉花棒觸 • 壓該偵測板之第二視窗内的基質； 步驟三，將㈣板置入第-順位的管瓶中（管瓶内為Hybridizati〇n buffer)，並將環蓋栓緊該管瓶，放至攝氏6()度恆溫供箱$分鐘後取出該偵測板； 步驟四，將侧板移至第二順位的管瓶中（管瓶内為muffer)，浸泡1〇 肇-秒清洗基質後取出； 步驟五’將_板移至第三順位的管瓶中（t瓶㈣SA_biQtin binding buffer) ’放入攝氏6〇度恆溫箱5分鐘後取出； 步驟六’將侧板移至第四順位的管瓶中（管瓶内為Wash buffer)，浸泡10 秒清洗基質後取出； 步驟七，將_板移至第五順位的管瓶中（管瓶内細则p bu版），放 入攝氏分鐘後取出；在此—步驟，若整峨進行正確，該侧板 ♦-之弟-視窗的基質將出現呈色反應；若樣品染病，將會於該侧板之第二 的基質出現呈色反應； 步驟八，將_板移至第六順位的f瓶中（管瓶嫌触_㈣，浸_ 秒清洗基質綱，此—清洗動作可讓實驗者更清楚嶋上述的呈色反應 必要時可職f烘乾明色踩。 … 關:呈，反應所代表之意義，以第七、八、九圖說明。若實驗步驟進行確 图正石梅細之第一視細的基質5〇會出現翻的呈色反應(如第七 回表不仏測成功，若無呈色反應，則表示檢測失敗(如第九圖），應重新採樣測 M316891 應有無，以判定此次檢測是否具有參 係著樣品是否染有疫病之結果，在第 試。據此，檢測者可由第一視窗lU呈色反 考價值。而第二視窗112的呈色反應，則關 七圖中第-Hfm、咖如⑽，絲養殖水施感染白點 病毒；若如第人圖，第二視窗112無呈色反應，則表示樣品無感染。M316891 VIII. New description: [New technical field] • The thief amplification method and the immunization method are related. In more detail, it is a rapid detection device for nuclear acid and immunoassay. [Prior Art] Nucleic acid amplification technology and immunological method have become one of the most important tools in the field of biology. In the field of clinical medical applications, the detection of pathogens, genetic diseases, etc. plays a pivotal role. _ - Fortune products detection of diseases as an example, introduce the known paste polymerase Lianhang (p (8) combined with the biochip detection process. First use PCR to amplify specific nucleic acid fragments, and then add the PCR amplified product to the hybrid reaction solution After injection into the wafer cassette, put it into the hybrid reaction box, 'mix for at least - hour at a specific ambient temperature (_ need to constantly shake to enhance the hybrid reaction), and pour off the liquid in the wafer after the hybridization is completed. Add the cleaning solution, pour off the cleaning solution and add a specific concentration of avidin-inducible phosphatase (SA-AP), react at room temperature for ten minutes, then pour off the liquid in the wafer, add the cleaning solution, and rinse off Add a specific concentration of the coloring agent (10)T/BCIp) and react at room temperature for ten minutes. Call the wafer again and then rinse with water. Observe whether the wafer has a color contrast or not. A disease. The above-mentioned detection methods 'whether it is test ship selection, dosage, process, operation mode, environmental control, etc., are all known by the experts in the form of "knowledge" and "theoretical", and the reagents used are used. And the hardware is also presented in the form of experimental equipment, and stored in the laboratory, which makes the detection method become very difficult to re-age, so that it must be carried out by specialized personnel, and the detection environment is limited to the experiment. In the room. These reasons have made it impossible for the testing work to be carried out by the maintainers themselves, which may cause delays in the immediate prevention and control of the disease. In addition, if the specialists do not fully comply with the standard 5 M316891, the test results may be seriously mis-processed for the test procedure, or the test procedure error is poor, but it is not easy to be noticed. [New content] The purpose of the above-mentioned complicated and difficult case is to convert the detection method of the rapid detection device into a simple and specific. - a nucleic acid method and a quick touch, x set, mainly includes: a detection plate having at least one base for presenting a specific test result, and eight, the soil can be a plastic piece, a glass, a nylon, One of the nitrocellulose membranes, the polyethylene sulphide membranes, and the like, wherein one side of the detection panel has at least one hollowed-out window through which the sample can contact the base f-ring cover and is coupled thereto. The detector board is stored in a tube _, and each tube is provided for the above-mentioned _ plate, and the bottle opening of the bottle can be fixed to the ring cover. Compared with the detection method described in the prior art, the present invention will detect a plurality of vials of a sequence, such as a reagent, a dosage, a flow, an operation mode, and the like, and the detection sample uses the detection plate as a carrier, and the detection sample is used as a carrier. The price test board can also present the test wire and have a success or loss of lying. In this case, the detection method __ is simple, fast, specific, and the correct rate is improved. Therefore, it can be carried out by the aquarists in the farm, which has immediate and effective significance for the detection and prevention of the disease. [Embodiment] As shown in the first, second and third figures, the rapid detection device for the nucleic acid amplification method and the immunological method of the present invention mainly comprises: a detecting plate 10 which defines a narrow and long plate shape in terms of a length and a width ratio, The detection plate 10 has at least one substrate 50 for presenting a specific detection result, and the substrate 50 may be a plastic sheet, a glass, a nylon film, a nitrocellulose film, a polyvinylidene fluoride film, or the like. The detecting plate 10 defines at least one hollow window 11 through which the sample can contact the substrate 50; M316891 - a ring cover 20 coupled to the top end of the detecting plate; a plurality of reagents stored in the vial 30 31. Each of the vials 30 can be received by the detecting plate 1 , and the bottle opening 32 of the vial can be fixed to the ring cover 20 . In the case of the case, the detecting board 1 is formed by a combination of a panel 12 and a bottom plate a. The panel 12 includes a columnar joint 14 at the top end, a substrate positioning slot on the back side, and a first window ni and a second window 112 that are hollowed out. A substrate 50 having a specific detection function is disposed in the substrate positioning groove 15. The bottom plate 13 is bonded to the back surface of the panel 12 to completely enclose the above-described substrate positioning groove 15, and the substrate 50 is fixed. The bottom plate 13 and the panel 12 are coupled to each other at opposite positions of the two to provide a plurality of protrusions 16 and recesses 17 which are tightly fitted to each other. As shown in the first, second, fifth and sixth figures, in the embodiment of the present invention, the ring cover 2 is provided with a reduction of the columnar joint portion 14 of the detecting plate 10, according to which the 2G is combined with the The top of the board 1 。. The joint portion of the splicing groove 21 and the columnar joint portion 14 is provided with a convex ring 24 and a ring groove 141' which are engageable with each other so that the cover 2 can be firmly coupled with the detecting plate 1 。. The ring cover 2 and the bottle mouth 32 of the vial are provided with threads 23 and 33 which can be screwed to each other. When the cross-plate 1 is placed in the vial 3, the ring cover 2 can be screwed. In the bottle π32. The top surface of the ring cover 20 is provided with a plate-like holding portion 23, the shape of which is convenient for fingers to hold, and the marking of the sample is more conveniently placed thereon. As shown in the fourth embodiment, in the embodiment of the present invention, the vial 30 is sealed by a sealing cover 34 to seal the internal reagent. If the reagent is a chemically stable chemical, it can be stored in the vial in a liquid state; If the reagent is not easy to store the active enzyme or the like, the end of the cold-drying preparation is stored in the bottom of the bottle, and a predetermined amount of water is added to activate it. The above-mentioned vials 3 are coded in the order of detection so that the examiners can take them in order. Take the case of whether the cultured aquatic products are infected with white spot virus, and how to use the rapid detection device in this case. M316891 - Steps - ' Recording instructions to take the water-like product of the scorpion water _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ • Press the substrate in the second window of the detection plate; Step 3, place the (4) plate into the first-order vial (Hybridizati〇n buffer in the vial) and fasten the ring cap to the vial. Remove the detection plate after 6 minutes Celsius at a constant temperature of 6 ° C. Step 4, move the side plate to the second-order vial (muffer in the vial), soak for 1 〇肇-sec to clean the substrate. Then take out; Step 5' Move the _ plate to the third-order tube (t bottle (four) SA_biQtin binding buffer) 'Put in the 6-degree Celsius incubator for 5 minutes and take it out; Step 6' Move the side plate to the fourth position In the vial (Wash buffer inside the tube), soak it for 10 seconds to clean the substrate and take it out; Step 7, move the _ plate to the fifth-order tube (in the tube bottle, pbu version), put it in Celsius Take out after a minute; in this step, if the whole sputum is correct, the side plate ♦-the younger brother-the window will appear a color reaction; If the sample is infected, a color reaction will appear on the second substrate of the side plate; Step 8. Move the plate to the f bottle of the sixth position (the bottle is suspected _ (four), dip _ seconds to clean the matrix, this - The cleaning action can make the experimenter more clear. The color reaction mentioned above can be used to dry the bright color when necessary. ... Off: Present, the meaning of the reaction, as illustrated in the seventh, eighth and ninth diagrams. The matrix 5〇 of the first-order micro-precision of the Orthodox sylvestris will appear to have a color reaction (if the seventh table is not successful, if there is no color reaction, it means the detection fails (as in the ninth figure). The M316891 should be re-sampled to determine whether the test has the result of whether the sample is infected with the disease, in the first test. According to this, the tester can be color-retested by the first window. In the color reaction of the window 112, the first-Hfm and the coffee (10) in the Fig. 7 are applied, and the silk culture water is infected with the white spot virus; if the first window 112 has no color reaction, the sample is free from infection.
。與先前技術所述之檢測方法相較，本案將檢測所需之試劑、用量、流程、 操作方式等製做成序列的若干管瓶，而檢職品則_侧板城具，且該摘 測板另可呈現檢驗結果以及具有_檢測為成功或失敗的指標。本案使檢測方 法變得鮮、快速、具體、且正轉提高，因此可以交由養殖㈣於養殖場中 自行實施，這對於疫病的檢測及防治具有即時而有效的意義。 、軸核砂-個最錄施舰朗，但齡此祕者能在獨離本案精 神”範可下做各種不同形式的改變。以上所舉實施例僅用以說明本案而已，非 用以限制本案之测。舉凡不違核精神所從事的種種修改錢化，俱屬本案 申凊專利範圍。 一 【圖式簡單說明】 奏圖為本案债測板的立體分解圖。 第二圖為本案偵測板的組合外觀圖。 第二圖為本案偵測板的組合剖視圖。 第四圖為本案依檢測所需步驟而序列設置的管瓶平面圖。 第五圖為本案偵測板與管瓶結合的示意圖。 第六圖為本案偵測板與管瓶結合的剖面圖。 第七圖為本案_板表示檢献似及具有疫病指標的呈色反應示意圖。 第八圖為本案細彳板表示檢測成似及未染疫標之反應示意圖。 9 M316891 弟九圖為本案偵測板表不檢測失敗的不意圖。 【主要元件符號說明】 10-偵測板 20-環蓋 11-視窗 21-樞接槽 111-第一視窗 22-螺紋 112-第二視窗 23-持取部 12-面板 24-凸環 13-底板 30-管瓶 14-柱狀結合部 31-試劑 141-環槽 32-瓶口 15-基質定位槽 33-螺、紋 16-凸柱 34-密封蓋 17-凹槽 50-基質. Compared with the detection method described in the prior art, the present invention will detect a number of vials of a sequence, such as reagents, dosage, flow, operation mode, etc., and the inspection products are _ side plate tools, and the test The board can also present inspection results and indicators with _ detection as success or failure. This case makes the detection method fresh, fast, specific, and forward-moving. Therefore, it can be administered to the farm (4) in the farm itself. This has immediate and effective significance for the detection and control of the disease. The axis of the core sand - one of the most recorded Shi Lang, but the age of this secret can be made in the spirit of the case alone can be made in various forms of change. The above examples are only used to illustrate the case, not to limit The test of this case. All kinds of modifications and moneys that are not in violation of the nuclear spirit are all in the scope of patent application in this case. A [simplified description of the figure] The picture is a three-dimensional exploded view of the debt test board of the case. The combined view of the test board. The second picture is a sectional view of the combination of the detection board of the case. The fourth picture is a plan view of the tube set according to the steps required for the detection. The fifth picture shows the combination of the detection board and the tube bottle. The sixth figure is a cross-sectional view of the combination of the detection plate and the tube bottle of the case. The seventh picture is a schematic diagram of the color reaction of the case indicating the presence of the disease and the disease indicator. The eighth picture shows the detection of the fine plate of the case. Schematic diagram of the reaction of the uninfected epidemic. 9 M316891 The younger figure is not intended to detect failure of the detection board. [Main component symbol description] 10-detection board 20-ring cover 11-window 21- pivot Slot 111 - first window 22 - thread 112 - second window 23 - holding portion 12 - panel 24 - convex ring 13 - bottom plate 30 - vial 14 - columnar joint 31 - reagent 141 - ring groove 32 - bottle mouth 15 - substrate positioning groove 33 - screw, 16-bump 34-sealing cap 17-groove 50-matrix