TW594002B - Fluid detecting chamber device of biosensor and its application - Google Patents

Abstract

The present invention provides a fluid detecting chamber device of biosensor and its application. The device includes a shell having a first inlet, a second inlet, a first outlet and a second outlet, and a piezoelectric quartz crystal arranged in the shell and having a first detecting face and a second detecting face. In addition, the device further includes two pipe pairs, including a first pipe pair and a second pipe pair, configured in the shell, wherein the first pipe pair is connected to the first inlet and first outlet to communicate with the first detecting face, and the second pipe pair is connected to the second inlet and second outlet to communicate with the second detecting face.

Description

594002 7706twf · doc / 〇〇6 A7 B7 V. Description of the invention (() The present invention relates to a fluid detection chamber device of a biosensor and its application, and particularly to a piezoelectric quartz crystal organism Sensor (Piezoelectric Quartz Crystal Biosensor) fluid detection chamber device and its application. Piezoelectric quartz crystal biosensor, also known as Quartz Crystal Microbalance (QCM), is a type of detection that has received much attention in recent years. Method. Because it has the advantages of simple principle, convenient operation, low cost, and instant response, it can be used in medical diagnosis, genetic testing, food analysis, and environmental monitoring. In addition, because it has many advantages, it can be applied It is also very extensive, so it may be possible to replace some traditional detection methods in the future. Piezoelectric Quartz Crystal Immunosensor is a new method of immunoassay. The principle of immunoassay is Affinity adsorbed ligand (usually antibody or antigen) immobilized on crystal electrode On the surface, and then inject the test object (sample). When the test object and the ligand on the electrode surface generate immunoaffinity adsorption, the weight of the crystal surface will increase, and then the frequency signal of the crystal will decrease by It is used as the basis for testing. There is a relationship between the change in quality and frequency (1): (Please read the note on the back? Matters before filling out this page) Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economy

AF

F

R

p dA

Am

F

R 2 ΝΑ

Am-(1) where △ F: frequency change amount (Hz) p Q: density of quartz (g / cm3) This paper size applies Chinese National Standard (CNS) A4 specification (210 X 297 mm) 594002 A7 B7 77 06twf doc / 006 V. Description of the invention (of) N: Frequency coefficient of quartz d: Thickness of quartz (cm) FR: Frequency of response (Hz) A: Area of adsorption (cm2) △ m: Mass absorbed on the wafer Change (g) the relationship between the frequency and the mass of the quartz crystal microbalance, as shown in the formula (1), when the frequency changes (AF), we can calculate the mass of the substance (△ m) attached to the wafer, and The material on the wafer is usually a very thin layer, and the cockroach is only a few hundred nanometers, so it is very small, so it can be applied to the quantitative detection of trace molecules. The traditional methods of immobilizing proteins (antibodies or antigens) on the surface of crystal electrodes are physical adsorption or chemical bonding methods. However, these methods are not only costly and complicated to produce. In addition, conventional quartz crystal biosensors use only a single detection surface of the quartz crystal for detection. Therefore, only a single test object can be detected at a time. Therefore, the object of the present invention is to provide a fluid detection chamber device of a piezoelectric biosensor, which is used to immobilize a protein (antibody or antigen) by an electropolymeric polymer method. Therefore, the protein can be embedded in different electrochemical conditions Quartz crystal biosensor to achieve the purpose of low cost and simple production. Another object of the present invention is to provide a biosensor detection system, which has the advantages of simple principle, convenient operation, low cost, and instant response, and also has a plurality of detection surfaces, thereby improving the sensitivity during detection. . 4 This paper size applies to China National Standard (CNS) A4 (21〇X 297 public love) ------------------- I ^ · IIII I-- (Please first Read the notes on the back and fill in this page) Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 594002 y A7 7706twf.doc / 006 B7 -------------- V. Description of the invention (> ) (Please read the precautions on the back before filling out this page) Another object of the present invention is to provide a biosensor detection system. Since it has a plurality of detection surfaces, it can detect multiple test objects in a single specimen. It not only reduces cost, but also has high sensitivity and simple operation. The invention provides a fluid detection chamber device for a biosensor. The device includes a casing having a first inlet, a second inlet, a first outlet, and a second outlet. A piezoelectric quartz wafer having a first detection surface and a second detection surface. In addition, it further includes two pipeline pairs arranged in the casing, and the two pipeline pairs include a first pipeline pair and a second pipeline pair. The first pipeline pair is connected to the first inlet and the first outlet and communicates with the first detection surface, so that a mobile phase flows from the first inlet to the first detection surface, and then flows out through the first outlet. The second pipeline pair is connected to the second inlet and the second outlet and communicates with the second detection surface, so that the mobile phase flows out from the first outlet, flows into the second detection surface through the second inlet, and then passes through the second Outflow. Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economics The present invention provides a detection system for a biosensor, which includes an injection device, a fluid detection chamber device, an oscillator circuit, and a frequency processing device for a test object. The injection device contains a sample to be tested. The fluid detection chamber device is in communication with the injection device. The fluid detection chamber device further includes a housing having at least two inlets and at least two outlets, and a piezoelectric quartz wafer arranged in the housing and having at least two detection surfaces opposite to each other. And at least two pipeline pairs, each pipeline pair having a first pipeline and a second pipeline, wherein the first pipeline is connected to a corresponding one of the inlets so that a mobile phase flows into the corresponding one One detection surface, the second pipeline is connected to one of the corresponding outlets, so that the five paper sizes are applicable to the Chinese National Standard (CNS) A4 (210 X 297 mm) 594002 A7 B7 7706twf · d〇c / 〇06 V. Description of the invention (bucket) (Please read the precautions on the back before filling out this page) The mobile phase then flows out of the corresponding one of the detection surfaces through one of the corresponding outlets. The oscillating circuit is coupled to the piezoelectric quartz wafer in the fluid detection chamber device, and is used to generate a frequency signal to the fluid detection chamber device, and then generates a frequency of the object to be measured from the fluid detection chamber device. The DUT frequency processing device is coupled to the oscillating circuit for calculating and displaying data of the DUT. Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economics The present invention proposes a detection method for a biosensor detection system. This method first provides a biosensor detection system. The system includes a fluid detection chamber device, an oscillating circuit and An injection device, wherein the fluid detection chamber device further includes a casing having at least two inlets and at least two outlets, a piezoelectric quartz wafer disposed in the casing and having at least two detection surfaces opposite to each other, and at least two pipeline pairs Each pair of pipelines has a first pipeline and a second pipeline, wherein the first pipeline is connected to a corresponding one of the inlets, so that a mobile phase flows into a corresponding one of the detection surfaces, and the second The pipeline is connected to a corresponding one of the outlets, so that the mobile phase flows out of the corresponding one of the detection surfaces through the corresponding one of the outlets. Next, the detection surface in the Jiejie fluid detection chamber device uses an electropolymeric polymer method to fix an affinity molecule (such as an antibody or an antigen) on the detection surface, and then uses an injection device to inject a sample to be tested into the fluid detection Chamber device, so that one of the test object in the test sample and the affinity molecule have an affinity reaction, and finally use the oscillation circuit of the detection system to detect the test object. The present invention utilizes constant voltage and variable voltage to electropolymerize conductive polymers to achieve the purpose of fixing affinity molecules (antibodies or antigens). Since this fixing method requires only one main step to complete, it is simpler and faster than general methods. 6 This paper size applies the Chinese National Standard (CNS) A4 specification (210 X 297 mm) 594002 A7 B7 7706twf. Doc / 00 6 V. Description of the invention (ί) The detection system of the biosensor of the present invention is a general improvement The flow cell of a common piezoelectric biosensor allows its mobile phase to pass through the multiple detection surfaces of the wafer, so that the measured object can get more than twice the immune response after the multiple detection surfaces. Response frequency while increasing its sensitivity. The detection system of the biosensor of the present invention fixes a plurality of different antibodies or antigens on a plurality of detection surfaces of a single wafer to simultaneously detect a plurality of different test objects in a single specimen. In order to make the above and other objects, features, and advantages of the present invention more comprehensible, a preferred embodiment is described below in detail with the accompanying drawings as follows: Brief description of the drawings: FIG. 1 It is a schematic cross-sectional structure diagram of the fluid detection chamber device of the biosensor of the present invention; FIG. 2 is a schematic diagram of the cross-section of I-Γ in FIG. 1; FIG. 3 is a schematic diagram of the detection system of the biosensor of the present invention; The figure is a chemical structure diagram of a commonly used electropolymerized polymer; Fig. 5 is an explanatory diagram of a detection profile of an electropolymerized polymer immobilized protein in Example 1 of the present invention; and Fig. 6 is an actual measurement of Example 1 of the present invention Standard curve diagram of human serum albumin antigen; Figure 7 is a diagram illustrating the detection profile of the same protein fixed on both sides of the crystal in Example 2 of the present invention; Figure 8 is the actual amount of one and two sides of Example 2 of the present invention Test for human blood 7 This paper size applies to Chinese National Standard (CNS) A4 (210 X 297 mm) ------------ installation -------- order · ---- ---- Line (Please read the notes on the back before filling this page) Intellectual Property Bureau of the Ministry of Economic Affairs Printed by the Employee Consumption Cooperative 594002 A7 B7 7706twf. Doc / 006 V. Description of the invention ((d) Standard curve diagram of albumin antigen; and Figure 9 is a detection section of the third embodiment of the present invention immobilized different proteins on both sides of the crystal Explanation of the diagram: 20: housing 211, 221: inlet 212, 222: outlet 24; L, 242, 243, 244: pipeline 200: mobile phase flow direction 22: loop pipe fitting 2: piezoelectric quartz Wafer 21: Metal electrode 31: Injection device 32 = Mobile phase 33: Fluid detection chamber 34: Oscillation circuit 35: Power supply 36: Computer 37: Pump 38: Waste liquid example shown in Figure 1, which is shown as Schematic diagram of the cross-section structure of the fluid detection chamber device of the invented biosensor; Figure 2, which is shown as the schematic diagram of Section Ⅰ_Γ in Figure 1. This paper size applies to China National Standard (CNS) A4 (210 X 297 mm) ) ------------ ^ Installation ·· ------- Order --------- Line »(Please read the notes on the back before filling this page) Economy Printed by the Consumer Cooperatives of the Ministry of Intellectual Property Bureau 594002 A7 B7 7706twf · doc / 006 V. Description of Invention (9) Please refer to In FIG. 1, the fluid detection chamber of the biodetector of the present invention includes a first inlet 211, a first outlet 212, a second inlet 221, a second outlet 222, and a housing 2. The first pipeline 241 connected to the first inlet 211, the second pipeline 242 connected to the first outlet 212, the third pipeline 243 connected to the second inlet 221, and the second pipeline connected to the second outlet 222, respectively. Four pipelines 244. A piezoelectric quartz wafer 2 is placed in the center of the casing 20, wherein the piezoelectric quartz wafer 2 has a first detection surface and a second detection surface. A metal electrode 21 (as shown in FIG. 2) is formed on the first detection surface and the second detection surface of the piezoelectric quartz wafer 2, respectively. The material of the metal electrode 21 is, for example, gold or silver. In addition, a loop pipe 22 is arranged between the first outlet 212 and the second inlet 221 to communicate the first detection surface and the second detection surface of the piezoelectric quartz wafer 2. In the fluid detection chamber of the biosensor of this embodiment, the flow direction of the object to be measured is shown by arrow 200. After a sample to be measured is injected from the first inlet 211, it enters the first pipeline 241. It then flows to the first detection surface of the piezoelectric quartz wafer 2 to perform the immune detection reaction, and then flows to the first outlet 212 through the second pipeline 242. Then, it flows into the second inlet 221 through the loop pipe 22, flows through the third pipeline 243, and flows to the second detection surface of the piezoelectric quartz wafer 2 for an immunological detection reaction, and then flows out of the fourth pipeline 244, The second outlet 222 flows out. In this embodiment, the characteristics of a quartz crystal microbalance are used for immunological detection, and the antibody or antigen to be tested is fixed on the detection surface of the piezoelectric quartz wafer 2 by means of an electropolymerized polymer immobilization method, and then the detection surface The upper metal electrode 21 detects the conjugated antigen or antibody at different concentrations. This Example 9 1 This paper size applies the Chinese National Standard (CNS) A4 specification (^ Nuogongfa) (Please read the note on the back? Matters before filling this page) Order --------- Ministry of Economics Printed by the Consumer Property Cooperative of the Intellectual Property Bureau 594002 7706twf.doc / 006 A7 B7 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs 5. Description of the invention (") The piezoelectric quartz chip 2 used is, for example, AT-cut 9MHz, its specifications The diameter of the quartz is 8mm, the diameter of the electrode is 4.5mm, and the thickness is 0,17mm. The device is based on Taiwan Taisheng Technology Co., Ltd. piezoelectric quartz crystal ultra-micro analyzer (model SB01B). Figure 3 'shows a schematic diagram of the detection system of the biosensor of the present invention. Please refer to FIG. 3. The detection system of the biosensor of this embodiment includes an injection device 31, a mobile phase 32, a fluid detection chamber 33, an oscillation circuit 34, a power supply 35, a computer 36, a pump 37, and Effluent tank 38. The fluid detection chamber 33 is designed for the fluid detection test in the first figure. An inlet of the fluid detection chamber 33 is connected to the injection device 31, and the injection device 31 is further connected to a mobile phase 32. The outlet of one of the fluid detection tests 33 is connected to a pump 37 to draw the flow direction 32 into the fluid detection chamber 33, and at the same time, bring the specimen in the injection device 31 into the fluid detection chamber 33. When the specimen enters the fluid detection chamber 33, antibodies or antigens on the detection surface of the piezoelectric quartz crystal therein will react. After that, devices such as the oscillating circuit 34 and the power supply 35 coupled to the piezoelectric quartz wafer are used to detect the test object on the piezoelectric quartz wafer. Afterwards, a computer 36 can be used to process the results of the analytical tests. The sample after the test is conveniently pumped to the waste liquid tank 38 by the pump 37. The steps of using the detection system of the biosensor of the present invention to perform detection analysis are as follows: First, (1) clean the electrode: immerse the metal electrode on the piezoelectric quartz wafer with 1 · 2M sodium hydroxide (NaOH) for 20 minutes , And use 1.2M hydrogen chloride (HC1) 10 This paper size is applicable to China National Standard (CNS) A4 specification (210 X 297 mm) ------------ installation -------- Order --------- line ίί (please read the precautions on the back before filling this page) 594002 A7 B7 77 06twf. Doc / 00 6 V. Description of the invention (1) Soak for 5 minutes, then use alcohol and Rinse with water to clean the metal electrode. (Please read the precautions on the back before filling in this page) After that, (2) Immobilize the protein: prepare electroplating solution for electropolymerized polymers, where the electropolymerized molecules are, for example, Polypyrrole, Polyaniline, Poly The chemical formula of Polythiophene, Poly (p-phenylene), Polyacetylene and its derivatives is shown in Figure 4. The electroplating solution of the electropolymerized polymer contains antibodies or antigens. Thereafter, the electropolymerized polymer is electroplated on a metal electrode on the quartz surface of the piezoelectric wafer in a variable voltage or constant voltage manner, and the electropolymerized polymer is embedded with an antibody or an antigen during polymerization. Then, the surface of the electrode was washed with phosphate buffer solution and water, and dried at room temperature. Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs. Electropolymeric polymer is a new material with electrical conductivity and reversible redox properties. It can also have special binding properties with metals and plastics. Electropolymerized polymers embed and immobilize proteins by taking advantage of their electrochemical synthesis and polymerization properties. When a crystal electrode is energized, its polymer monomers begin to polymerize and combine to form a positively charged film on the electrode surface. The protein to be immobilized is placed in the body solution, so that when the protein is negatively charged, the protein will be embedded in the polymer to complete the immobilization of the protein. In this embodiment, the method of using an electropolymerized polymer to fix proteins is also used. The main monomer used is Pyrrole, which is a good conductive tube molecular material, which can be used under different electrochemical conditions. For example, under constant voltage, constant current, or variable voltage conditions, biological functions such as Entrap proteins are embedded while controlling polymerization to achieve one-step protein immobilization. The use of electropolymeric polymers in the preparation of piezoelectric crystal biosensors can achieve the goals of low cost and simple fabrication. 11 National Standard (CNS) A4 specification (21G X 297 public love) of this New Zealand standard 594002 A7 B7 7706twf. Doc / 006 5. Description of the invention (…) Next, carry out (3) Testing method: the pressure after drying The electric quartz wafer is placed in the fluid detection chamber 33 in FIG. 3. Buffer was used as the mobile phase 32 and its flow rate was fixed. After the frequency of the disturbance circuit 34 is stabilized, a blocking agent is first injected by the injection device 31 as a mask for the detection surface of the piezoelectric quartz wafer. Then, the conjugate antigen or antibody of the test object is injected for immunological detection. At this time, the concentration of the test object can be determined by the frequency change of the oscillation circuit 34. After the detection of one specimen, a weak base or a weak acid buffer can be used as a desorbent to wash out the conjugated antigen or antibody adsorbed on the detection surface of the piezoelectric quartz wafer, and then proceed to the next specimen Of detection. In the detection system of the biosensor of the present invention, metal electrodes 21 are formed on both detection surfaces of the piezoelectric quartz wafer. Therefore, the fluid detection chamber device is designed as a double-sided flow device (as shown in FIG. 1). ). In addition, a loop pipe piece 22 of about 3 ml or more is added to the flowing pipeline to delay the detection time of the other detection surface of the piezoelectric quartz wafer. This will increase the frequency of its detection response and increase its detection sensitivity. In addition, two different antigens or antibodies can be immobilized on two detection surfaces to detect two different conjugated antibodies or antigens in a single specimen. The present invention will be described in detail with the following examples: Example 1 Figure 5 is not an explanatory diagram of a detection profile of a protein immobilized by an electropolymerized local molecule in Example 1 of the present invention. After the piezoelectric quartz wafer was cleaned, a Pyrrole plating solution containing an antibody (AbA) or an antigen (AgA) was prepared. Then, Esau ring volts from 0 volts to 12 paper standards are applicable to China National Standard (CNS) A4 (210 X 297 mm) ---------- I -------- Order- -------- (Please read the note on the back? Matters before filling out this page) Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 594002 A7 B7 7706twf.doc / 006 V. Description of the Invention (") (Please (Read the precautions on the back before filling in this page.) 1 volt for 20 seconds, or 0.25 volts to 1 volt for 60 seconds at a constant voltage. Electropolymerize the polymer by electroplating, so that the polymerized polymer is packed at the same time as polymerization. Immobilized antibodies. Then, the metal electrode on the detection surface of the piezoelectric stone central wafer was rinsed with a buffer solution, rinsed with water, and then dried at room temperature. Thereafter, the dried piezoelectric quartz wafer is placed in the fluid detection chamber 33 in Fig. 3. Phosphate Buffered Saline (PBS) was used as the mobile phase 32, and the flow rate was fixed at 125 μl / πη. After the frequency of the oscillating circuit 34 is stabilized, first cover the unreacted aldehyde groups on the metal electrode surface of the detection surface with 0.1M glycine, and then cover the surface of the metal electrode with 0.5% casein solution. Vacancy. After that, please refer to Figure 5 and inject 500μ1 of different concentrations of the test substance (AgA) or (AbA) for immunodetection. Here, when the antibody (AbA) is added to the plating solution, the injected test substance Then the antigen (AgA). Conversely, when the antigen (AgA) is added to the plating solution, the test substance injected is the antibody (AbA). After that, the frequency change of the oscillating circuit 34 is recorded. At this time, the quantity of the measured object can be obtained from the frequency change conversion. Printed in Figure 6 of the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs, which is a standard curve diagram of the actual measurement of human serum albumin antigen according to the first implementation of the present invention. Please refer to Fig. 6, which uses 0.4 volts for 60 seconds to plate an electropolymerized polymer to fix human serum albumin antibody (Anti-Human serum albumin (Anti-HSA)) on the electrode surface of a piezoelectric quartz wafer. Then, the results of the standard curve of measuring the adsorption of human serum albumin (HSA) were measured. Among them, the horizontal axis in Figure 6 is human blood. 13 The paper size is applicable to the Chinese National Standard (CNS) A4 (210 X 297 mm) " 594002 A7 B7 7706twf. Doc / 006 5. Description of the invention (α ) The concentration of albumin antigen, and the vertical axis is the magnitude of the measured oscillation frequency. Example 2 is shown in Figure 7, which illustrates the detection profile of the same protein fixed on both sides of the crystal in the second embodiment of the present invention. Illustration. Please fix the antibody (AbA) or antigen (AgA) on the electrode surfaces of the two detection surfaces of the piezoelectric quartz wafer according to Figure 7. Rinse the electrode surface with buffer solution and water and leave it at room temperature. dry. After drying, the piezoelectric quartz wafer is placed in the fluid detection chamber 33 of Fig. 3. After that, a phosphate physiological buffer solution was used as the mobile phase 32, and the flow rate was fixed to ι25 μ1 / πηη. After the frequency of the oscillating circuit 32 is stabilized, first cover the unreacted aldehyde groups on the electrode surface with 0.1M Glycine, then cover the vacancies on the electrode surface with 0.5% Casein solution, and then Inject 500μ1 of the test substance (AgA) or (AbA) for immunodetection. If the antibody (AbA) is immobilized on the electrode surface, the test solution injected is the antigen (AgA). Conversely, when the antigen (AgA) is added to the plating solution, the test substance injected is the antibody (AbA). Here, since the electrodes or antigens on both sides of the piezoelectric quartz wafer are fixed with the detection antibody or antigen, the frequency change of the response can be increased, and the sensitivity can be improved. Figure 8 shows the standard curve of the actual measurement of human serum albumin antigen on one and double sides of the second embodiment of the present invention. Please refer to Fig. 8. In Fig. 8, when a single-sided and double-sided piezoelectric quartz wafer is used, an anti-Human serum albumin (Anti-HSA) is fixed to the piezoelectric quartz wafer. The surface of the electrode is applicable to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) in 14 paper sizes -------------------- Order --- ------ line (Please read the note on the back? Matters before filling out this page) Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs 594002 A7 B7 77 0 6twf. Doc / 00 6 V. Description of the invention (丨 >) Measure the result of the standard curve of the adsorption of human serum albumin (HSA). In Fig. 8, the horizontal axis is the concentration of human serum albumin antigen, and the vertical axis is the magnitude of the oscillation frequency. As can be seen from Fig. 8, the frequency response of detection using a double-sided piezoelectric quartz wafer is approximately double the frequency response of detection using a single-sided piezoelectric quartz wafer. Example 3 As shown in FIG. 9, it is not an explanatory diagram of a detection section for immobilizing different proteins on both sides of a piezoelectric quartz wafer according to the third embodiment of the present invention. Please refer to FIG. 9. After the antibodies A (AbA) and B (AbB) are respectively fixed on the electrodes on the two detection surfaces of the piezoelectric quartz wafer, the surface of the piezoelectric quartz wafer is washed with buffer solution and water. Dry at room temperature. After drying, the piezoelectric quartz wafer is placed in the flow chamber 33 of the double-sided detection system. Thereafter, a phosphate physiological buffer solution was used as the mobile phase 32, and the flow rate was fixed at 125 μ1 / πήη. After the frequency of the oscillating circuit 34 is stabilized, unreacted aldehyde groups on the electrode surface of the detection surface are covered with o.im glycine, and then the vacancies on the electrode surface are covered with a 0.5% Casein solution. Then inject a fixed amount of the test object (AgA) or (AgB), and then by the frequency response of the two test objects, you can know the concentration of the test object (AgA) or (AgB), respectively. This embodiment uses a fluid detection chamber suitable for a double-sided piezoelectric quartz wafer as a detailed description. The present invention is more applicable to a piezoelectric quartz wafer with more than two detection faces. To sum up, the present invention has the following advantages: 1. The present invention uses a constant voltage and variable voltage method to electropolymerize conductive polymers to achieve the purpose of fixing proteins. Since this fixing method only requires 15 paper sizes, it is applicable to China National Standard (CNS) A4 specification (210 X 297 mm) -------------..------- Order- -------- Line »(Please read the note on the back? Matters before filling out this page) Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 594002 A7 77Q6twf.doc / 006 _ ^ _ V. Description of the Invention (Μ ) Fix protein in one main step, simpler and faster than normal methods. 2. The biosensor detection system of the present invention is to improve the fluid chamber of a common piezoelectric biosensor, so that its mobile phase can pass through a plurality of detection surfaces of a piezoelectric quartz wafer, so that the measured object passes through a plurality of detection surfaces. After detecting the immune response on the surface, the response frequency can be more than doubled, and the sensitivity can be improved. 3. The biosensor detection system of the present invention is a method of fixing a plurality of different antibodies or antigens on a plurality of detection surfaces of a single wafer to simultaneously detect a plurality of different test objects in a single specimen. Although the present invention has been disclosed as above with a preferred embodiment, it is not intended to limit the present invention. Any person skilled in the art can make some changes and retouch without departing from the spirit and scope of the present invention. The scope of protection of the invention shall be determined by the scope of the attached patent application. (Please read the precautions on the back before filling out this page) Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 16 This paper size applies to China National Standard (CNS) A4 (210 X 297 mm)

Claims (1)

594002 A8 B8 77 0 6twf. Doc / 006 ^ L) 〇. Patent application scope 1. A fluid detection chamber device of a biosensor, comprising: a housing having a first inlet, a second inlet, A first exit (please read the precautions on the back before filling out this page) and a second exit; a piezoelectric quartz wafer arranged in the housing, the piezoelectric quartz wafer has a first detection surface and A second detection surface; and two pipeline pairs arranged in the housing, the two pipeline pairs including a first pipeline pair and a second pipeline pair, wherein the first pipeline pair is connected to the The first inlet is connected to the first outlet and communicates with the first detection surface, so that a mobile phase flows from the first inlet to the first detection surface, and then flows out through the first outlet, and the second pipeline The pair is connected to the second inlet and the second outlet and communicates with the second detection surface, so that the mobile phase flows out from the first outlet, flows into the second detection surface through the second inlet, and then Flow out through this second outlet. 2. The fluid detection chamber device of the biosensor as described in item 1 of the scope of the patent application, further comprising a circuit connected between the first outlet and the second inlet. Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs 3. The fluid detection chamber device of the biosensor as described in item 1 of the scope of patent application, wherein the first detection surface and the second detection surface further include separate configurations There is a metal electrode. 4. The fluid detection chamber device of the biosensor as described in item 3 of the patent application scope, wherein the material of the metal electrode is selected from gold and silver. 5. The fluid detection chamber device of the biosensor as described in item 1 of the patent application scope, wherein the mobile phase is selected from a gas phase substance and a liquid phase substance. 17 This paper size applies to China National Standard (CNS) A4 (21 × χ297 mm) Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 594002 B8 7706twf.doc / 006 6. Scope of patent application 6. — Kind of fluid testing room The device includes: a casing having at least two inlets and at least two outlets; a wafer disposed in the casing, the wafer having at least two detection surfaces opposite to each other; and at least two pipe pairs disposed in the casing Among them, each of the pipeline pairs has a first pipeline and a second pipeline, and the first pipeline is connected to a corresponding one of the inlets so that a mobile phase flows into the corresponding One of the detection surfaces, the second pipeline is connected to the corresponding one of the outlets, so that the mobile phase flows out of the corresponding one of the outlets. 7. The fluid detection chamber device according to item 6 of the patent application scope, further comprising at least one circuit, so that the mobile phase is communicated from one of the pipeline pairs to one of the outlets connected to the other by One or more of the inlets to which the pipes are connected. 8. The fluid detection chamber device according to item 6 of the patent application scope, wherein the wafer includes a piezoelectric quartz wafer. 9. The fluid detection chamber device according to item 6 of the scope of the patent application, further comprising a metal electrode disposed on each of the detection surfaces. 10. The fluid detection chamber device according to item 9 of the scope of patent application, wherein the material of the metal electrode is selected from gold and silver. 11. The fluid detection chamber device according to item 6 of the scope of the patent application, wherein the mobile phase is selected from a gas phase substance and a liquid phase substance. 12. A detection system for a biosensor, comprising: an injection device containing a sample to be measured; a fluid detection chamber device communicating with the injection device, wherein the flow of 18 paper standards is applicable to the country of China Standard (CNS) A4 specification (210 X 297 mm) ------------------- Order --------- line (Please read the note on the back first Please fill in this page again for details) 594002 B8 7706twf.doc / 006 ^ VI. Patent application body testing room device includes = one housing with at least two inlets and at least two outlets; (Please read the precautions on the back before filling this page ) A piezoelectric quartz wafer is arranged in the casing, and the piezoelectric quartz wafer has at least two detection surfaces opposite to each other; at least two pipeline pairs are arranged in the casing, and each of the pipeline pairs has A first pipeline and a second pipeline, the first pipeline is connected to the corresponding one of the inlets, so that a mobile phase flows into the corresponding one of the detection surfaces, the second pipeline Is connected to the corresponding one or more outlets, so that the mobile phase is passed from the corresponding one or more outlets Output; an oscillating circuit coupled to the piezoelectric quartz wafer in the fluid detection chamber device for generating a frequency signal to the fluid detection chamber device, and then generating a frequency of the object to be measured from the fluid detection chamber device And a frequency processing device of the object to be tested, which is coupled to the oscillating circuit for calculating and displaying data of the object to be tested. 13. The detection system for a biosensor as described in item 12 of the scope of patent application, wherein the device for frequency processing of the object under test includes a computer. Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 14. The detection system of the biosensor as described in item 12 of the scope of patent application, which further includes a pump connected to one of these outlets to extract the mobile phase to The mobile phase is caused to flow in the fluid detection chamber device. 15. The detection system of the biosensor as described in item 12 of the scope of patent application, further comprising at least one circuit, so that the mobile phase is communicated from one of the pipelines to one of the outlets connected to One of the other inlets to which the other pipelines are connected. 16. Inspection of the biosensor as described in item 12 of the scope of patent application 19 This paper size applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) 594002 7706twf.doc / 006 ^ Six, the scope of patent application The measurement system further includes a metal electrode disposed on each of the detection surfaces. (Please read the precautions on the back before filling this page) 17. The detection system of the biosensor as described in item 16 of the patent application scope, wherein the material of the metal electrode is selected from gold and silver. 18. The detection system for a biosensor as described in item 12 of the patent application scope, wherein the mobile phase is selected from a gas phase substance and a liquid phase substance. 19. A method for detecting a biosensor detection system, comprising the following steps: providing a biosensor detection system including a fluid detection chamber device, an oscillating circuit and an injection device, wherein the fluid detection chamber device comprises: a A housing with at least two inlets and at least two outlets; a piezoelectric quartz wafer disposed in the housing, the piezoelectric quartz wafer having at least two detection surfaces opposite to each other; printed by at least the consumer cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs Two pipeline pairs are arranged in the casing, and each of the pipeline pairs has a first pipeline and a second pipeline, and the first pipeline is connected to a corresponding one of the inlets. A mobile phase is caused to flow into the corresponding one or more detection surfaces, and the second pipeline is connected to the corresponding one or more outlets, so that the mobile phase flows out from the corresponding one or more outlets again; The detection surfaces in the fluid detection chamber device; using an electropolymeric polymer method to fix an affinity molecule on the detection surfaces; using the injection device to inject a sample to be tested to A fluid detection chamber device to make one of the test object and the affinity molecule produce a affinity of 20 paper standards for China National Standard (CNS) A4 (210 X 297 mm) 594002 A8 B8 7706twf.doc / 〇〇6 C8 -__ D8 VI. Patent application scope and response; and the oscillation circuit of the detection system to detect the test object. 20. The detection method for a biosensor detection system as described in item 19 of the patent application scope, wherein the affinity molecule includes an immunodetection antibody. 21. The detection method for a biosensor detection system as described in item 20 of the scope of the patent application, wherein the test object is a co-antigen corresponding to one of the antibodies for immunodetection. 22. The detection method for a biosensor detection system as described in item 19 of the patent application scope, wherein the affinity molecule includes an immunodetection antigen. 23. The detection method for a biosensor detection system as described in item 22 of the scope of the patent application, wherein the test object is a conjugated antibody corresponding to one of the immunodetection antigens. 24. The detection method for a biosensor detection system as described in item 19 of the scope of patent application, further comprising fixing a plurality of affinity molecules on the detection surfaces. 25. The detection method of the biosensor detection system according to item 24 of the scope of the patent application, wherein the affinity molecules are fixed to analyze a plurality of test objects in the sample. 26. The detection method of a biosensor detection system as described in item 19 of the scope of application for a patent, wherein the electropolymerized polymer method uses a variable voltage method to convert the ~~~ ^ electropolymerized polymer molecules to the test Surface. 27. The detection method of the biosensor detection system as described in item 26 of the patent application scope, wherein the variable voltage method is performed with a sweep ring voltage of 0 volts to 1 volt for 20 seconds. This paper size applies to China National Standard (CNS) A4 (210 X 297 mm) (Please read the precautions on the back before filling this page) Order --------- Line-Intellectual Property Bureau, Ministry of Economic Affairs Printed by Employee Consumer Cooperative 594002 A8 B8 C8 D8 7706twf.doc / 006 Printed by the Consumer Property Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs. Scope of patent application. 28. As described in the patent application scope of the biosensor detection system test method 26 , Where the electropolymeric polymer is selected from polypyrrole (please read the notes on the back before filling in this page) (Polypyrrole), polyaniline, polythiophene, poly (p-styrene) (Poly (p-phenylene)), Polyacetylene and its derivatives. 29. The detection method of a biosensor detection system as described in item 19 of the scope of patent application, wherein the electropolymerized polymer method uses a certain voltage method to electroplate an electropolymerized polymer onto the detection surface. 30. The detection method of the biosensor detection system according to item 29 of the scope of the patent application, wherein the constant voltage method is performed at 0.25 volts to 1 volt for 60 seconds. 31. The detection method for a biosensor detection system as described in item 29 of the patent application scope, wherein the electropolymeric polymer is selected from the group consisting of Polypyrrole, Polyaniline, and Polythiophene , Poly (p-phenylene K-Poly (p-phenylene)), Polyacetylene and its derivatives. 32. The detection method of the biosensor detection system according to item 19 of the scope of the patent application, wherein the detection surfaces further include a metal electrode each configured. 33. The method for detecting a biosensor detection system according to item 19 in the scope of the patent application, wherein the mobile phase is selected from a gas phase substance and a liquid phase substance. 34. The detection method for a biosensor detection system as described in item 33 of the patent application scope, wherein the liquid phase substance includes a phosphate buffered saline (PBS). 22 This paper size applies to China National Standard (CNS) A4 (210 x 297 mm)