M393516 五、新型說明： 【新型所屬之技術領域】 本創作係關於一種能使可攜式即時定量微生物檢驗設備的溫度 升降穩定又快速，縮短單次循環所需時間，提高反應循環速率，同時 減少溫控調整時之耗能的qPCR加熱冷卻模組。 【先前技術】 隨著螢光偵測技術的快速進步，在9〇年代中期開發出即時定 量聚合酶連鎖反應(Real-time PCR，qPCR)偵測系統，一面複製 DNA，一面同時偵測螢光訊號之操作機轉，有效克服了傳統pcR 技術容易因為後處理造成實驗室污染，而發生的偽陽性問題解決 分子生物在基目定量上的_，qPCR在各項生摘域所能應 用的範圍很廣泛，同時又能夠提供快速、經濟且精準的效能，例如 基因表現的研究、感染性病原菌的檢測、食品的檢測、新藥開發的 樂效評估以及單-基因遺魅病檢測（SNp)及環境微生物的檢測 等°即時定量聚合酶連鎖反應能有效的降低檢測時間並具備高專一 性的優點，朗在公共衛生或是職場魏衛生JL，能朗快速且有 效進仃環境監測及檢測的目標，進而姐的預防疫情的發生或減緩 疫障的擴散’目此若能發展可在作業環境麟操作之體積小、重量 輕的可攜切狀量聚麵連鎖反舰備麟疾病之感染與擴散、 生物攻擊與缝防衛等醫護防疫需求有所助益。㈣定量聚合酶連 M393516 鎖反應，藉由控制溫度來讓DNA變性（d e n a t u r e)、黏合（a n n e a 11 n g)及延伸（extension)，並藉由具專一性的引子 及探針（_e)標稀複狀顧此，放大並从料定的生物 細胞或微生物之DNA片段，藉此定量出環境中微生物之含量，因 此即時定量聚合酶連鎖反應設備中負責控制溫度的加熱降溫模組， 成為該設備反應效能最主要的決定因素。 總體而言，習知的㈣定量聚合酶連鎖反應設備之加熱时模 組多為體積大、溫控反應速率慢且溫度穩定度不佳，因此該設備多 為大而重的桌上型設備，防疫在健現場採樣後必須帶回實 驗室中才能進行檢測’如此—來延遲了出具報告的時間，並且增加 了採樣後贿處_舰’若保存條料#或鮮錯誤，都將使檢 測結果發生嚴重誤差。 本案創作人餅上述M qPCR域降溫模組所衍生的各項缺 點，乃虽思加以改良創新’並經多年潛心研究後，終於研發完成本 件qPCR加熱降溫模組。 【新型内容】 _作之目的在於提供一種qPCR加熱降溫模組，透過此模組 達到精準溫控與快速加熱降溫的目的，以降低侧反應所需的循環 時間，同時減少溫控調整時之能源使用，逹到輕量化與低耗能，可 ^ ;體積小、重I輕的可攜式qpCR，利於隨身攜帶，能立即在 M393516 現場檢測致病性微生物’避免人為採樣保存處理之風險，有助於衛 生防疫與勞工衛生安全防護措施之施行。為達上述目的之qPCR加 熱降溫模組，包括一控制單元，係由電源供應器、ΡΠ)控制器、繼 電器和熱電偶所組成；一加熱單元，係由兩支卡式加熱器所組成； 一冷卻單元’係由致冷晶片與散熱鰭片所組成；以及一測試單元。 【實施方式】M393516 V. New description: [New technical field] This creation is about making the temperature rise and fall of the portable real-time quantitative microbial testing equipment stable and fast, shortening the time required for a single cycle, increasing the reaction cycle rate, and reducing QPCR heating and cooling module for energy consumption during temperature control adjustment. [Prior Art] With the rapid advancement of fluorescence detection technology, a real-time quantitative polymerase chain reaction (qPCR) detection system was developed in the mid-1990s to simultaneously detect DNA while replicating DNA. The operation of the signal machine effectively overcomes the traditional PCR technology, which is easy to cause laboratory pollution due to post-processing, and the false positive problem occurs to solve the quantitative problem of molecular biology in the base. The range of qPCR that can be applied in various fields Wide range, while providing fast, economical and accurate performance, such as research on gene expression, detection of infectious pathogens, detection of food, evaluation of the evaluation of new drug development, and single-gene enchantment detection (SNp) and environment Microbial detection, etc. The real-time quantitative polymerase chain reaction can effectively reduce the detection time and has the advantage of high specificity. It can be used in public health or workplace health JL, and can quickly and effectively enter the target of environmental monitoring and detection. In addition, the sister's prevention of the epidemic or the spread of the epidemic will be reduced. If it can be developed, it can be operated in the working environment. Portability cut chain-like weight polyethylene surface anti-ship equipment Lin infection and spread of the disease, the seam attack and defense and other biological health care and epidemic prevention needs help. (4) Quantitative polymerase and M393516 lock reaction, by denaturation, adhesion (annea 11 ng) and extension by controlling temperature, and by using specific primers and probes (_e) In view of this, the DNA fragments of the biological cells or microorganisms are amplified and quantified to quantify the content of microorganisms in the environment, so that the heating and cooling module responsible for controlling the temperature in the polymerase chain reaction device is instantly quantified, and the reaction efficiency of the device is obtained. The most important determinant. In general, the conventional (4) quantitative polymerase chain reaction equipment heating module is mostly bulky, temperature-controlled reaction rate is slow and temperature stability is not good, so the equipment is mostly large and heavy desktop equipment, The epidemic prevention must be brought back to the laboratory for testing after being sampled at the health site. [This - delays the time for issuing the report, and increases the post-sampling bribe _ ship's if the stock # or fresh error will make the test result A serious error has occurred. In this case, the creators of the above-mentioned M qPCR domain cooling module derived from the defects are thought to be improved and improved. After years of painstaking research, the qPCR heating and cooling module was finally developed. [New content] _ The purpose of _ is to provide a qPCR heating and cooling module, through which the purpose of precise temperature control and rapid heating and cooling is achieved, so as to reduce the cycle time required for side reaction and reduce the energy during temperature control adjustment. Use, light weight and low energy consumption, can be small, heavy I light portable qpCR, easy to carry, can immediately detect pathogenic microorganisms in the M393516 on-site to avoid the risk of human sampling preservation, there is Helps the implementation of health and epidemic prevention and labor health safety protection measures. The qPCR heating and cooling module for the above purpose comprises a control unit consisting of a power supply, a controller, a relay and a thermocouple; and a heating unit consisting of two card heaters; The cooling unit 'is composed of a cooled wafer and heat sink fins; and a test unit. [Embodiment]
請參閱圖一，本創作的qPCR加熱冷卻模組，主要包括有：一控 制單元(3)，由一個電源供應器(31)、一個PID控制器(32)、兩個繼電 器(34、35)以及一個熱電偶(33)組成；一加熱單元(1)，由兩支8〇^的 卡式加熱器(11)組成；一冷卻單元(2)，由一個185W的致冷晶片（22) 和一個散熱鰭片（21)組成；以及一測試單元(4)。Referring to Figure 1, the qPCR heating and cooling module of the present invention mainly comprises: a control unit (3), a power supply (31), a PID controller (32), and two relays (34, 35). And a thermocouple (33); a heating unit (1) consisting of two 8 卡 card heaters (11); a cooling unit (2) consisting of a 185 W chilling wafer (22) and A heat sink fin (21); and a test unit (4).
將含有待測樣本的玻片置於設備中的測試單位(4)，進行即時定 量聚合酶連鎖反應如圖二所示’設備中的加熱冷卻模組首先啟動卡式 加熱器(11)將溫度由室溫加熱至94°C，而PID控制器(32)則控制卡式 加熱器(11)使溫度維持於94°C以進行DNA變性作用（denature);接 著卡式加熱器(11)停止動作，同時致冷晶片（22)啟動使溫度下降，當 溫度接近60°C時，則致冷晶片（22)停止運作，PID控制器(32)將溫 度維持於60°C，以進行DNA黏合作用（annealing);隨後啟動卡式 加熱器(11),而PID控制器(32)則控制卡式加熱器(11)使溫度維持於 72°C，以進行DNA延伸作用（extension);最後卡式加熱器(11)再度 5 M393516 啟動使溫度回到94°C，並重複循環直到完成所設定的循環次數為止。 在本實施例中，本加熱降溫模組能達成一個2.7kg的可攜式即時 定量聚合酶連鎖反應設備之溫度穩定度為±i.rc，一個反應循環時間 為116秒’能於作業環境中即時測出致病微生物之濃度，有利於勞工 防護之施行。 本創作所提供之加熱冷卻模組，與其他習用技術相互比較時，更 具有下列之優點，可應用於輕量型qPCR，利於隨身攜帶至待測現場 即時定量環境中微生物含量，且可精確並快速調控反應溫度，不僅避 免採樣後樣本保存之風險’更節省反應時間與所需耗能，如表一所示： 表一Place the slide containing the sample to be tested in the test unit (4) of the device, and perform the instantaneous quantitative polymerase chain reaction as shown in Figure 2. The heating and cooling module in the device first activates the card heater (11) to set the temperature. Heating from room temperature to 94 ° C, while the PID controller (32) controls the card heater (11) to maintain the temperature at 94 ° C for DNA denaturation; then the card heater (11) stops The action, while the cooling chip (22) is started to lower the temperature, when the temperature is close to 60 ° C, the cooling chip (22) stops operating, and the PID controller (32) maintains the temperature at 60 ° C for DNA bonding. Annealing; then the card heater (11) is activated, and the PID controller (32) controls the card heater (11) to maintain the temperature at 72 ° C for DNA extension; the final card The heater (11) is again activated by 5 M393516 to bring the temperature back to 94 ° C and repeat the cycle until the set number of cycles is completed. In this embodiment, the heating and cooling module can achieve a 2.7kg portable real-time quantitative polymerase chain reaction device with a temperature stability of ±i.rc and a reaction cycle time of 116 seconds' in the working environment. Instant detection of the concentration of pathogenic microorganisms is conducive to the implementation of labor protection. The heating and cooling module provided by the present invention has the following advantages when compared with other conventional technologies, and can be applied to lightweight qPCR, which is convenient for carrying the microbial content in the instant quantitative environment to be tested, and can be accurately Rapid regulation of reaction temperature not only avoids the risk of sample storage after sampling, but also saves reaction time and required energy consumption, as shown in Table 1: Table 1
Company/Instrument Dimensions (WxDxH cm) Weight (kg) Heating Rate (°CVs) Cooling Rate (°C/s) qPCR加熱降溫模組 13x24x18 2.7 1.67 2.29 Hppendorf^Mastercycler® family 26 x 41 x 27 12.4 3 2 Eppendor£^Mastercycler® pro 26x41.5x37 18.5 4 3 Eppendorf^Mastercycler® ep realplex 26 x 41 x 39.6 7 4 3 Peqlab/Primus Thermocycler 22.5 x 25 x 28 6.3 2 2 Applied Biosystems/ GeneAmp® PCR System 9700 30 x 41 x 26 8.7 1 1 Applied Biosystems/2720 Thermal Cycler 21x36x22 N/A 1 1 Applied Biosystems/Veriti® 384-Well Thermal Cycler 24.5 x 23.7 x 48.5 11.4 3.1 3.1 ESCO/Swift MaxPro Thermal Cycler BLC5 30.6 x 38.6 x 29.5 10 1.8 1.8 上述詳細說明係針對本創作可行實施例之具體說明，惟該實施例 並非用以限制本創作之專利範圍，凡未脫離本創作技藝精神所為之等 6 M393516 效實施或變更，均應包含於本案之專 综。所述，本案不但在輕量化、時間節省與溫度控制上確屬創 新’並能較習用物品增進上述多項功效，應已充分符合新雛及進步 性之法定創作專利要件，爰依法提出_請，懇請貴局核准本件創作 專利申凊案’以勵創作，至感德便。 【圖式簡單說明】 圖一係為本創作實施例之方塊示意圖 圖二係為本創作實施例之使用示意流程圖 【主要元件符號說明】Company/Instrument Dimensions (WxDxH cm) Weight (kg) Heating Rate (°CVs) Cooling Rate (°C/s) qPCR Heating and Cooling Module 13x24x18 2.7 1.67 2.29 Hppendorf^Mastercycler® family 26 x 41 x 27 12.4 3 2 Eppendor£ ^Mastercycler® pro 26x41.5x37 18.5 4 3 Eppendorf^Mastercycler® ep realplex 26 x 41 x 39.6 7 4 3 Peqlab/Primus Thermocycler 22.5 x 25 x 28 6.3 2 2 Applied Biosystems/ GeneAmp® PCR System 9700 30 x 41 x 26 8.7 1 1 Applied Biosystems/2720 Thermal Cycler 21x36x22 N/A 1 1 Applied Biosystems/Veriti® 384-Well Thermal Cycler 24.5 x 23.7 x 48.5 11.4 3.1 3.1 ESCO/Swift MaxPro Thermal Cycler BLC5 30.6 x 38.6 x 29.5 10 1.8 1.8 The detailed description of the present invention is not intended to limit the scope of the patents, and the implementation or modification of the 6 M393516 effect, which is not included in the spirit of the creative spirit, should be included in the case. . As mentioned, this case is not only innovative in terms of weight reduction, time saving and temperature control, but also can enhance the above-mentioned multiple functions compared with the conventional items. It should have fully complied with the new and progressive statutory creation patent requirements, and submitted it according to law. I would like to ask you to approve the creation of a patent application for this article. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a block diagram showing the use of the present embodiment. FIG. 2 is a schematic flow chart showing the use of the present embodiment.
1加熱單元 11卡式加熱器 2冷卻單元 21散熱鰭片 22致冷晶片 3控制單元 31電源供應器 32 PID控制器 33熱電偶 34固態繼電器1 35固態繼電器2 4測試單元1 heating unit 11 card heater 2 cooling unit 21 heat sink fin 22 cooling chip 3 control unit 31 power supply 32 PID controller 33 thermocouple 34 solid state relay 1 35 solid state relay 2 4 test unit