201226896 六、發明說明： 【發明所屬之技術領域】 本發明係一種微生物或細胞的檢測系統及其方法，特 別係指一種將微生物或細胞代謝特性應用於電化學檢測方 式的微生物或細胞的檢測系統及其方法。 【先前技術】 在一般的微生物或細胞檢測系統中，大多是將一微生 物或細胞培養於一特定成份之培養基或培養液中，使得此 微生物或細胞得以大量生長，再利用人工計數、光學性或 籲檢測特定微生物或細胞生長代謝產物含量的方法來推算微 生物或細胞的總數。 最大可能數法（Most Probable Number)是一種常見的傳 統檢測方法，其係將培養有菌體的菌液進行稀釋為檢液， 並吸取1毫升檢液接種於已裝有9毫升硫酸月桂酸胰化蛋 白月示培養液中（LST_Lauryl Sulfate Tryptose Br〇th)培養， 並，察疋否產生培養液是否產生氣體，來初步鑑別培養液 籲巾疋否有大腸杯菌群’若為陽性反應需再進行後續以特 f培養基進行培養觀察_是否具有特定生化反應甚至 疋進订染色及顯微鏡觀察以確認生成菌株之種類，十分耗201226896 VI. Description of the Invention: [Technical Field] The present invention relates to a microorganism or cell detection system and method thereof, and more particularly to a microorganism or cell detection system for applying microbial or cellular metabolic properties to an electrochemical detection method And its method. [Prior Art] In a general microbial or cell detection system, a microorganism or a cell is cultured in a medium or a culture medium of a specific component, so that the microorganism or the cell can be grown in a large amount, and then manually counted, optically or A method of detecting the content of a specific microorganism or cell growth metabolite is used to estimate the total number of microorganisms or cells. The Most Probable Number is a common traditional detection method. It dilutes the cultured bacteria solution into a test solution, and takes 1 ml of the test solution to inoculate 9 ml of lauric acid sulfate. Cultured in the protein culture medium (LST_Lauryl Sulfate Tryptose Br〇th), and check whether the culture medium produces gas, to initially identify the culture medium, and whether there is a large intestinal flora [if it is a positive reaction, then Carry out subsequent culture observation with special f medium_Whether there is a specific biochemical reaction or even a microscopic observation and microscopic observation to confirm the type of strain produced, which is very expensive
另外有些商業化套組則是利用特定菌株的生理生化特 性或是利用特定染$姑 A 、 “ 技5來進行菌株的鑑別，但是這些方 法都僅能進行定性的八此 ,.^ y 的刀析’並無法確知實際菌落數量，因 此仍需後續的典1 & 員妁培養來進行定量性的分析。 201226896 【發明内容】 本發明的主要目的在於提供一種微生物或細胞的檢測 系統，此系統是-種電化學檢測系統，其係包括有一反應 槽、-電極單元、1置於反應槽的待測主體和_處理器， 其中反應槽内設置有供應待測主體生長的環境，此環境係 由-培養液或-特^化學成份組成，電極單元具有一感測 端’感測端係$ s於反應槽内i可供肖冑主體生長或吸 附’處理器係連結於電極單元，並包括有一電壓單元、一 控制單元和-讀取單元，錢單元係連接於電極單元並供 給-電壓於電極單it，jt電Μ為-固定電壓或是—可變電 壓，控制單元係連接於電壓單元以控制輸出至電極單元的 電壓輸出值或區間’讀取單元係連結於電極單元以定時讀 取電極單元上的電化學參數變化狀況，這些電化學參數包 括有電流值、頻率值或阻抗值。 本發明的另一目的在於提供一種微生物或細胞的檢測 方法，其係利用在一微生物或細胞培養環境中設置一電 極，以檢測在通入一可變電壓並來回偵測所誘發之電流變 化，或在通入一特定電壓的情況下，而培養基成份包含會 誘發產生電解之成份，藉測量一定期間所誘發之電流、頻 率或阻抗等變化，來達到檢測特定微生物或細胞是否存在 及整體數量的目的，其中前述電壓施加時間係持續〇至12 小時，且以連續性施加或間歇性施加，而可變電壓的電壓 值介於-50至50伏特間。 在本發明中，當微生物或細胞培養環境中存在有所欲 檢測之目標微生物（或細胞）或是非目標微生物（或細胞）之其 201226896 他微生物時’電極所感測到的變化也會有所不同，例如（i) ^培養丨兄中含有一特定待測微生物或細胞時，電極所檢 測到的電流、頻率或阻抗值將持續變大，舉例來說，當目 標微生物若在培養環境中大量快速產酸，則會造成培養環 境的酸鹼值（pH值）快速下降，並因此造成目標微生物脫離 電極表面，或是產生對目標微生物具傷害性之成份（包含微 生物代謝產物或電解產物，或結合兩者作用）而造成目標微 生物的破壞使細胞物質流出細胞外，進而引起電流、頻率 φ或阻抗值產生顯著變化；若是在通入一固定電壓予電極的 清況下，則因電化學反應將產生相當大之電流變化；又若 通入可變電壓予電極後則會產生一相當大之電流峰，藉由 债測誘發產電流變化的時間或產生電流峰所須時間，即可 區別並判定所檢測樣品中特定微生物之含量，並藉以區別 出目標微生物的濃度；（2)當培養環境中不含特^微生物或 細胞，或是微生物或細胞無法在此培養環境下生長時，電 極所檢測到的電流、頻率或阻抗將會維持在一恆定的狀 籲態；（3)培養環境内含有非目標微生物，且此非目標微生物 也可於此培養環境中進行生長時，微生物代謝產生離子之 變化僅會造成些可忽略之電流、頻率或阻抗之變化’但是 因非目標微生物菌體也會逐漸吸附於電極’而造成電極界 面電容提高而阻礙電子傳遞，因此造成電流、頻率或阻抗 出現微幅下降的狀況。 綜上所述，依據本發明之系統及方法，本發明係結合 下列反應，（1)微生物產酸而使培養環境pH下降進而造成 菌體電荷改變而脫離電極而造成訊號的變動，⑺結合低壓 201226896 電場與特定培養基成分，使電極附近產生特定成分而造成 特定微生物產生缺陷造成細胞内物質滲出，將可快速區別 出特定微生物或細胞的濃度或是總微生物或細胞數，並可 更進一步簡單且快速地檢測分析出樣品中之特定菌種或細 胞、總菌數或細胞的數量。 【實施方式】 以下，即以實施例說明本發明的微生物或細胞的檢測 系統及其方法。 請參考第1圖所示，本發明的檢測系統包括有一反應 槽1、-電極單元2、一設置於反應槽的待測主體3和一處 理器4’其中反應槽内設置有供應待測主體生長的環境此 環境係由一培養液或一特定化學成份組成，電極單元具有 一感測端，感測端係設置於反應槽内並可供待測主體生長 或吸附’處理器係連結於電極單元，並包括有_電壓單元、 -控制單元和一讀取單元，電壓單元係連接於電極單元並 供給-電壓於電極單元，此電壓為―固定電壓或是一可變 電塵，控制單元係連接於電壓單元以控制輸出至電極單元 的電墨輸出值或區Fa1 ’讀取單㈣連結於電極單元以 讀取電極單元上的電化學參數變化狀況，這些電化學參數 包括有電流值、頻率值或阻抗值。 在-利用本發明檢測大腸桿菌的實施例中，係使用平 、、白金電極做為電極早兀’反應槽中添加有一微生物培養 液，此微生物培養液中添加有硫酸月桂酸鈉、⑽鹽:氣 化納及奴源等選擇性藥劑，並保持為、 一雷懕於雷炻留-以 『心驗值，在施加 電壓於電極^的狀況下’大腸桿菌群會呈現帶有負電 201226896 荷的狀態並因此往電極的正極方向移動而吸附於電極表 面，當吸附於電極的微生物數量增加時，會使得電壓單元 施予電極單元所誘發之電流因微生物吸附而受阻並呈現微 巾田下降的狀況，但是因為大腸桿菌在此一培養液的環境下 會快速且大量產生酸性物質，並使菌體轉成帶正電荷而脫 離電極表面，加以電壓使培養液產生電化學變化，其電解 成分於酸性酸鹼值（低pH值）下，在此加乘作用會造成大腸 杯菌裂解而流出細胞產物，在一實施例中係施加一可變電 φ壓並以一固定速率進行掃描時，於初培養數小時電極所誘 發之電流因微生物逐漸吸附而下降，但隨大腸桿菌之生 長，微生物逐漸脫離電極及菌體破損流出呼吸酵素等成份 而大幅上升，檢測電極誘發產生電流峰所需時間即可用此 方法選擇性檢測樣品中之大腸桿菌濃度；在另一實施例中 係施加以一固定電壓，則大腸桿菌會在特定時間產生大幅 電流上升，檢測電極誘發產生電流所需時間即可用此方法 選擇性檢測樣品中之大腸桿菌濃度。In addition, some commercial kits use the physiological and biochemical characteristics of specific strains or use specific dyeing, and the identification of strains, but these methods can only be qualitative, and the knife of .^ y It is impossible to ascertain the actual number of colonies, so the subsequent routine 1 & 妁 culture is still needed for quantitative analysis. 201226896 SUMMARY OF THE INVENTION The main object of the present invention is to provide a microorganism or cell detection system, the system The invention relates to an electrochemical detection system, which comprises a reaction tank, an electrode unit, a main body to be tested and a processor disposed in the reaction tank, wherein the reaction tank is provided with an environment for supplying the growth of the main body to be tested. Composed of - broth or - chemical composition, the electrode unit has a sensing end 'sensing end system $ s in the reaction tank i can be used for growth or adsorption of the schematic body attached to the electrode unit, and includes There is a voltage unit, a control unit and a reading unit. The money unit is connected to the electrode unit and supplies a voltage to the electrode unit it, and the jt is a fixed voltage or a variable voltage. The unit is connected to the voltage unit to control the voltage output value or the interval outputted to the electrode unit. The reading unit is coupled to the electrode unit to periodically read the electrochemical parameter change condition on the electrode unit, and the electrochemical parameters include the current value. , frequency value or impedance value. Another object of the present invention is to provide a method for detecting microorganisms or cells by using an electrode in a microbial or cell culture environment to detect a variable voltage and detect it. Measure the induced current change, or in the case of a specific voltage, and the composition of the medium contains components that induce electrolysis, by measuring changes in current, frequency or impedance induced during a certain period of time, to detect specific microorganisms or The purpose of the presence and overall number of cells, wherein the aforementioned voltage application time is continued for up to 12 hours, and is applied continuously or intermittently, and the voltage value of the variable voltage is between -50 and 50 volts. In the microbe or cell culture environment, there is a target microorganism (or cell) to be detected. Or the non-target microorganism (or cell) of its 201226896 his microbes, the changes sensed by the electrodes will also be different, for example (i) ^ when the cultured sputum contains a specific microorganism or cell to be tested, the electrode is detected The current, frequency or impedance value will continue to increase. For example, if the target microorganism rapidly produces a large amount of acid in the culture environment, the pH value of the culture environment will decrease rapidly, and thus the target microorganism will be caused. Deviation from the surface of the electrode, or the production of components harmful to the target microorganism (including microbial metabolites or electrolysis products, or a combination of both) causes destruction of the target microorganism, causing the cell material to flow out of the cell, thereby causing current, frequency φ or The impedance value changes significantly; if a fixed voltage pre-electrode is applied, a considerable current change will occur due to the electrochemical reaction; if a variable voltage is applied to the electrode, a considerable amount will be generated. The current peak can be distinguished and judged by the time when the change of the current is induced by the debt test or the time required to generate the current peak. Measuring the content of specific microorganisms in the sample, and thereby distinguishing the concentration of the target microorganism; (2) when the culture environment does not contain special microorganisms or cells, or the microorganisms or cells cannot grow in the culture environment, the electrodes are detected The current, frequency or impedance will be maintained at a constant state; (3) the non-target microorganisms in the culture environment, and the non-target microorganisms can also grow in this culture environment, and the microbial metabolism produces ion changes. It will only cause negligible changes in current, frequency or impedance 'but the non-target microbial cells will gradually adsorb to the electrode', causing the electrode interface capacitance to increase and hinder electron transfer, thus causing a slight increase in current, frequency or impedance. The situation of decline. In summary, according to the system and method of the present invention, the present invention combines the following reactions: (1) the microbial acid production causes the pH of the culture environment to decrease, thereby causing the change of the cell charge to cause the signal to change from the electrode, and (7) combining the low voltage. 201226896 The electric field and the specific medium composition make the specific components in the vicinity of the electrode cause the defects of specific microorganisms to cause the exudation of intracellular substances, which can quickly distinguish the concentration of specific microorganisms or cells or the total number of microorganisms or cells, and can be further simple and simple. Quickly detect and analyze the specific strain or cell, total bacterial count, or number of cells in the sample. [Embodiment] Hereinafter, a detection system and method for a microorganism or a cell of the present invention will be described by way of examples. Referring to FIG. 1, the detection system of the present invention comprises a reaction tank 1, an electrode unit 2, a main body 3 to be tested disposed in the reaction tank, and a processor 4' in which a supply subject to be tested is disposed in the reaction tank. Environment for growth This environment consists of a culture solution or a specific chemical component. The electrode unit has a sensing end. The sensing end is disposed in the reaction tank and can be used for growth or adsorption of the body to be tested. The unit includes a voltage unit, a control unit and a reading unit. The voltage unit is connected to the electrode unit and supplies a voltage to the electrode unit. The voltage is “fixed voltage or a variable electric dust, and the control unit is Connected to the voltage unit to control the output value or the area Fa1 of the output to the electrode unit. The reading unit (4) is coupled to the electrode unit to read the change of the electrochemical parameter on the electrode unit, and the electrochemical parameters include the current value and the frequency. Value or impedance value. In the embodiment in which the Escherichia coli is detected by the present invention, a microbial culture solution is added to the reaction tank using a flat, platinum electrode as an electrode, and the sodium sulphate sodium sulfate and (10) salt are added to the microbial culture solution: Selective agents such as gasification and slaves, and keep it as a Thunder, and a Thunder will be left with a "heart test value, under the condition of applying voltage to the electrode ^", the E. coli group will exhibit a negative charge 201226896 The state is thus moved toward the positive electrode of the electrode to be adsorbed on the surface of the electrode. When the number of microorganisms adsorbed to the electrode is increased, the current induced by the voltage unit being applied to the electrode unit is hindered by microbial adsorption and the micropod field is lowered. However, because E. coli can rapidly and in large quantities produce acidic substances in the environment of the culture medium, and the bacteria are converted into positively charged and separated from the surface of the electrode, the voltage is applied to electrochemically change the culture solution, and the electrolytic component is acidic. At pH (low pH), the multiplication will cause the C. cerevisiae to lyse and shed the cell product. In one embodiment, a cell is applied. When the voltage is φ and is scanned at a fixed rate, the current induced by the electrode is decreased due to the gradual adsorption of microorganisms in the initial culture for several hours. However, with the growth of Escherichia coli, the microorganism gradually deviates from the electrode and the bacteria are broken and the respiratory enzymes are discharged. With a large increase, the time required for the detection electrode to induce a current peak can be used to selectively detect the concentration of E. coli in the sample; in another embodiment, a fixed voltage is applied, and E. coli will generate a large current at a specific time. Ascending, the time required for the detection electrode to induce a current can be used to selectively detect the concentration of E. coli in the sample.
實施例一 在本實施例中，係說明在施與一可變電壓的情況下， 利用本發明的檢測系統及方法進行微生物的定性和定旦Embodiment 1 In this embodiment, the detection system and method of the present invention are used to perform qualitative and fixed determination of microorganisms under the application of a variable voltage.
本實施例中’係將-電極放置於-疑似具有大腸里桿菌 產氣菌株的肉桂酸胰化蛋白腺培養液asT_iauW tryPt〇Se broth)中’並給^該電極丨至_丨伏特的電壓值，以 每分鐘50毫伏特（mV/min)的掃描速率進行檢測，並記錄電 極上所發生的物理或化學性變化結果。 請參考第2圖所示 電極置入培養液時，因為菌體 201226896 在正常培養基環境下均帶負電’所以在一外加低壓電場 下’菌體便會順著正極方向移動並吸附於電極表面，以施 加之可變電壓而電壓以一固定速率掃描時，電極所誘發之 電流因微生物逐漸吸附而下降。 再請參考第3A至3C圖所示，當電極置入含大腸桿菌 (五.coh)之培養槽時之循環伏安圖譜’該循環伏安圖譜在培 養第3小時時，觀察到尖峰電流（標線7)，其係由於大腸桿 菌表面出現損傷，而流出如細胞色素等電化學活性成份之 故。通入電壓時將因流出如細胞色素等電化學活性成份具 有傳導電子之作用而使電流變大，但由於流出如細胞色素 本身之氧化還原電位相當低，因此在通入可變電壓後因 此電極表面所誘發之電流將隨在細胞色素等在其氧化電位 之電壓時會有一相當大之電流變化，隨培養時間增加（如標 線8)’微生物會開始破裂並釋出大量細胞成份至培養環^ 中’而導致電流大幅變化。藉由本發明之方法所得之電壓 電流變化圖譜可以藉由偵測低電壓誘發之大幅電流變化， 即可區別並判定所偵測之微生物是為大腸桿菌或非大腸桿 菌。並進-步由线電流產生時間之快慢，決定大腸桿菌 之濃度，若非大腸桿菌培養基僅因微生物代謝產生離子濃 度變化，因此僅獲得些微的誘發電流，藉由本發明之方法 所得之循環伏安圖譜法，可鑑定為樣品中是否含有大腸桿 菌。 實施例 在本實施例中’係說明在施與__定電壓（例如幻伏）的 情況下’利用本發明的檢測系統及方法進行微生物的定姓 201226896 和定量。 本實施例與實施例一大致相同，其最大的差別在於本 實施例中係施加一定電壓於電極上，請參考第4圖所示， 其中標線9表示培養液中含有大腸桿g之電流變化曲線， 標線ίο纟示培養液中不含有大腸桿菌但是還有產氣桿菌 之電流變化曲線，另外，若是在培養液中添 加不同菌種，例如沙門氏菌、酵母菌、枯草桿菌、鏈黴菌、 礼酸桿菌時’電極所檢測到的電流並不會有顯著改變，因 此本發明方法確實對於不同菌種具有鑑別的能力。 實施例三 若是將本發明方法與一般平板計數法（plate count)相 較，可以得到如下表1之結果。 表1、數法和本發明方法之計數結果比較表 平板計數法 本發明方法 --(單位:每毫升的菌落生成單位對數傕） 0 0 0 0 0 0 0 0 1.7 2.3 4.35 4.16 4.98 5.0 5.06 5.0 5.00 5.0In the present embodiment, 'the electrode is placed in the cinnamic acid-suppressing protein gland culture medium asT_iauW tryPt〇Sebroth which is suspected to have a strain of Escherichia coli and the voltage is 丨 丨 丨 volt. The detection was performed at a scan rate of 50 millivolts per minute (mV/min) and the results of physical or chemical changes occurring on the electrodes were recorded. Please refer to the electrode shown in Figure 2 when the culture solution is placed, because the cell body 201226896 is negatively charged in the normal medium environment, so the bacteria will move along the positive electrode direction and adsorb on the electrode surface under a low voltage electric field. When the voltage is applied at a fixed rate with a variable voltage applied, the current induced by the electrode is lowered by the gradual adsorption of microorganisms. Referring again to Figures 3A to 3C, the cyclic voltammogram of the electrode when placed in a culture tank containing Escherichia coli (f. coh). The cyclic voltammogram was observed at the 3rd hour of culture, and a peak current was observed ( Marking line 7), which is due to the damage of the surface of E. coli, flows out of electrochemically active components such as cytochromes. When the voltage is applied, the current is increased by the action of the electron-active component such as cytochrome. However, since the redox potential of the cytochrome itself is relatively low, the electrode is supplied after the variable voltage is applied. The current induced by the surface will change with a considerable current when the cytochrome or the like is at the voltage of its oxidation potential. As the culture time increases (such as the line 8), the microorganism will begin to rupture and release a large amount of cellular components to the culture ring. ^中' causes a large change in current. The voltage-current variation map obtained by the method of the present invention can distinguish and determine whether the detected microorganism is Escherichia coli or Escherichia coli by detecting a large current change induced by a low voltage. The progress of the step-by-step line current determines the concentration of E. coli. If the E. coli culture medium only changes the ion concentration due to microbial metabolism, only a slight induced current is obtained, and the cyclic voltammetry method obtained by the method of the present invention is used. Can be identified as whether the sample contains E. coli. EXAMPLES In the present embodiment, the description of the microorganisms was carried out using the detection system and method of the present invention in the case where a predetermined voltage (e.g., illusion) was applied. This embodiment is substantially the same as the first embodiment, and the biggest difference is that in the present embodiment, a certain voltage is applied to the electrode, please refer to FIG. 4, wherein the line 9 indicates the current change of the large intestine rod g in the culture solution. Curve, the line ίο纟 shows the current curve of E. coli but not A. coli in the culture solution. In addition, if different strains are added to the culture solution, such as Salmonella, yeast, Bacillus subtilis, Streptomyces, and In the case of acid bacilli, the current detected by the electrode does not change significantly, so the method of the present invention does have the ability to identify different species. Example 3 If the method of the present invention is compared with a general plate count, the results of Table 1 below can be obtained. Comparison of Counting Results of Table 1, Number Method and Method of the Invention Table Tablet Counting Method The method of the present invention--(Unit: logarithm of unit of colony per ml) 0 0 0 0 0 0 0 0 1.7 2.3 4.35 4.16 4.98 5.0 5.06 5.0 5.00 5.0
由表中可以看出，本發明方法所檢測得到㈣數計算 結果與習用之平板計數法十分近似，卻可 ，_ u大幅減少培養 和進行各項鑑別實驗所需的時間。 實施例四 201226896 為了解菌落濃度與產生電流變化之相關性，本實施例 中確認不同濃度菌落會產生電流變化的時間，情參見第5 圖所示，標線u和12分別代表不同培養溫度之結果，由圖 中可以看出，本發明的檢測方法應用於大腸桿菌時，可以 在37t的培養溫度下，獲得良好之菌落漠度和產生電流變 化關係’其相關係數（R2)為0.9672。 綜上所述可知，大部分之傳統檢測法均需要經過一定 時間的培養後在對於特定菌株進行個別的鐘定試驗，其皆 至少皆須花費48小時才能知道檢驗結果為陰性，若要確認 陽性反應’則可能需要再花7_10天的時門 # θ .^ 化⑶天的時間，若是以固體培 養基内，亦需在培養24-48小時後，再以營光或呈色法來進 行大腸桿菌陽性判定，相當耗費人力及與時間；而習用之 之導電檢測法係利用微生物代謝產生離子濃度變化引發導 電度之變化來檢測微生物，當菌數需達到1〇6(單位生成數/ 毫升）才會有大幅訊號變化，但其僅能檢測微生物之濃度， 並無法判定微生物種類，檢測結果亦會受到培養 影響。 本發明則透過施加電Μ及透過調整培養成份使微生物 生長並於特定階段破裂及訊號轉換，可快速且準確地檢測 大腸桿菌及大腸桿g之濃度，非但選擇性高於^導電檢 測等系統’而且縮短檢測時間，&外，本發明之方法不似 過去電流式感測系統必需添加電子傳遞媒質或誘發生成電 子傳遞媒質的物質來完成檢測，又不似過去方法，無法區 分^腸桿g與其它微生物，此外，按本發明之方法可快速 測定、具高敏感度、成本效益方面亦具有優點且不會如傳 201226896 統檢測法般耗費人力及與時間。 【圖式簡單說明】 第1圖係本發明微生物或細胞的檢測系統之示意圖。 第2圖係利用本發明微生物或細胞檢測方法所檢測得 在施與一可變電壓下一般微生物吸附於電極上時之電流變 化情形。 第3 A圖係利用本發明微生物或細胞檢測方法檢測大腸 桿菌之一實施例中，於第〇和2小時所檢測得之電流電壓 變化情形。 籲 第3B圖標線7係利用本發明微生物或細胞檢測方法之 一實施例中，於第3小時所檢測得之電流電壓變化情形。 第3C圖標線8係利用本發明微生物或細胞檢測方法之 一實施例中，於第4小時所檢測得之電流電壓變化情形。 第4圖係說明大腸桿菌和產氣桿菌在本發明檢測方法 下電流變化和時間關係圖。 第5圖係說明大腸桿菌在不同培養溫度下利用本發明 方法所檢測得到菌落數和電流變化時間關係圖。 響 【主要元件符號說明】 1反應槽 2電極單元 3待測主體 4處理器 第〇 j時電極所檢測得之電流電壓變化曲線 第2 J時電極所檢測得之電流電壓變化曲線 7第3小時電極所檢測得之電流電壓變化曲線 201226896 8第4小時電極所檢測得之電流電壓變化曲線 9在施加一定電壓於一檢測大腸桿菌之培養環境且具 有大腸桿菌時，培養環境之電流變化曲線 10在施加一定電壓於一檢測大腸桿菌之培養環境但不 具有大腸桿菌時，培養環境之電流變化曲線 11大腸桿菌在本發明方法下施與一定電壓並在37<t培 養下，菌落濃度與產生電流變化時間的關係圖 12大腸桿菌在本發明方法下施與一定電壓並在45<t培 養下，菌落濃度與產生電流變化時間的關係圖As can be seen from the table, the calculation results of the (four) number detected by the method of the present invention are very similar to the conventional plate counting method, but the _u greatly reduces the time required for the cultivation and the various identification experiments. Example 4 201226896 In order to understand the correlation between colony concentration and current generation change, in this example, it is confirmed that different concentrations of colonies will cause current change time, as shown in Fig. 5, the marks u and 12 respectively represent different culture temperatures. As a result, it can be seen from the figure that when the detection method of the present invention is applied to Escherichia coli, a good relationship between colony inversion and current generation can be obtained at a culture temperature of 37 t, and the correlation coefficient (R2) is 0.9672. In summary, most of the traditional detection methods require individual incubation tests for specific strains after a certain period of incubation, all of which take at least 48 hours to know that the test results are negative, to confirm positive The reaction 'may take another 7-10 days of time. # θ . ^ (3) days, if it is in a solid medium, it is also necessary to culture for 24-48 hours, then camping or coloring method to carry out E. coli The positive judgment is quite labor-intensive and time-consuming; while the conventional conductivity detection method uses the change of the ion concentration caused by the microbial metabolism to induce the change of the conductivity to detect the microorganism, when the number of bacteria needs to reach 1〇6 (unit generation number/ml). There will be a large signal change, but it can only detect the concentration of microorganisms, and cannot determine the type of microorganisms. The test results will also be affected by the culture. The invention can rapidly and accurately detect the concentration of Escherichia coli and the large intestine rod g by applying electrolysis and adjusting the culture component to cause the microorganism to grow and rupture and signal conversion at a specific stage, which is not only selective than the system of conductive detection and the like. Moreover, the detection time is shortened, and the method of the present invention does not require the addition of an electron transport medium or a substance that induces the generation of an electron transport medium to complete the detection in the past current sensing system, and does not resemble the past method, and cannot distinguish the gut g. In addition to other microorganisms, in addition, the method according to the invention can be quickly determined, has high sensitivity, is cost-effective, and has advantages and is not as labor-intensive and time-consuming as the 201226896 method. BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a schematic view showing a detection system of a microorganism or a cell of the present invention. Fig. 2 is a view showing changes in current when a general microorganism is adsorbed on an electrode by applying a variable voltage by using the microorganism or cell detecting method of the present invention. Fig. 3A shows the change in current and voltage detected in the first and second hours in one of the examples of E. coli using the microorganism or cell detection method of the present invention. 3B icon line 7 is a change in current and voltage detected in the third hour using an embodiment of the microorganism or cell detecting method of the present invention. The 3C icon line 8 is a case where the current and voltage detected in the fourth hour are changed by the embodiment of the microorganism or cell detecting method of the present invention. Fig. 4 is a graph showing the relationship between current change and time in Escherichia coli and Aerogenophilus under the detection method of the present invention. Fig. 5 is a graph showing the relationship between the number of colonies and the time of change of current detected by E. coli at different culture temperatures by the method of the present invention. [Representation of main component symbols] 1 Reaction tank 2 electrode unit 3 Current and voltage change curve detected by the electrode of the main body 4 processor to be tested at the second stage J. The current and voltage change curve 7 detected by the electrode at the 2nd J hour Current and voltage change curve detected by the electrode 201226896 8 Current and voltage change curve detected by the electrode at the 4th hour of the electrode 9 When a certain voltage is applied in a culture environment for detecting Escherichia coli and Escherichia coli is present, the current change curve 10 of the culture environment is When a certain voltage is applied to a culture environment for detecting Escherichia coli but no Escherichia coli, the current curve of the culture environment is 11. Escherichia coli is subjected to a certain voltage under the method of the present invention, and the concentration of the colony and the current are changed under the condition of 37 < t culture. Time relationship Figure 12 Escherichia coli applied a certain voltage under the method of the present invention and the relationship between the colony concentration and the change time of the generated current under 45 < t culture
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