Chip-based reagent box for biological testing and its detection method

Abstract

The present invention provides a chip-based reagent box for biological testing and a detection method thereof. The detection device includes a flow path guide layer and is stacked with a substrate to form a main body, and a reaction area is provided on a part of a pair of electrodes on the substrate, and a receptor that binds to a detection target is applied to the reaction area. The flow path guide layer has an opening installed at the same position as the reaction area, and the receptor is exposed to the main body at the opening, and a micro flow path is formed between the flow path guide layer and the substrate. The detection method includes steps of dropping, washing, and detection, and the sample liquid is directly dropped into the reaction area from the opening to react with the receptor. After the reaction time, a washing liquid is added to the sample liquid and dropped into the reaction area, which is permeated into the micro flow path by capillary action and discharged, and then the test liquid is dropped into the reaction area to detect the sample. In this method, the sample detection can be completed without applying external force, and the detection efficiency can be improved.

Description

[Technical Field]

[0001] The present invention provides a testing device, and in particular, a chip-based reagent box for biological testing and a testing method thereof. [Background technology]

[0002] In conventional biochip testing devices, a receptor for binding to a test target is applied to a reaction area, which is installed in the testing device and not exposed to the outside. During testing, an operator collects an appropriate amount of sample droplet from a biological fluid (e.g., blood) and drops it into the inlet of the reaction area. An external power device applies force (e.g., pressurized gas) to transport the sample into the reaction area. The sample comes into contact with the receptor and undergoes a reaction, after which testing is performed and an analysis result is obtained. However, when applying external pressure to the sample using an external power device, the sample is easily damaged by the pressure, which can lead to inaccurate analysis results.

[0003] In another conventional biochip testing device, a sample liquid is dropped into the inlet of a reaction area, and the sample liquid is guided to the reaction area by capillary action, and after contact with the receptor, testing is performed to obtain an analysis result. However, the process of guiding the sample liquid to the reaction area takes time, and the reaction begins after the sample liquid is guided to the reaction area, so the testing time is delayed and it is difficult to effectively improve the testing efficiency. Summary of the Invention [Problem to be solved by the invention]

[0004] The inventor has developed a chip-based reagent box for biological testing and a testing method thereof that can efficiently complete sample testing without requiring external driving force. [Means for solving the problem]

[0005] Accordingly, a first aspect of the present invention provides a chip-based reagent box for biological testing, comprising: a main body, the main body being provided with a pair of electrodes, and a reaction region being formed in part of the pair of electrodes; a substrate on which a receptor that binds to a detection target is applied in the reaction region; and a flow-guiding layer provided on top of the substrate, the flow-guiding layer having an opening located at the same position as the reaction region, the receptor being exposed to the outside of the main body through the opening, allowing a sample liquid to be directly dropped into the reaction region and react with the receptor; a micro-channel communicating with the reaction region being present between the main body and the substrate, and a washing liquid being added to the sample liquid after the reaction and permeating into the micro-channel and then being discharged.

[0006] In one embodiment, the flow-guiding layer includes a hydrophilic layer having capillary action and a non-absorbent hydrophobic layer, the hydrophobic layer is disposed between the substrate and the hydrophilic layer, the openings penetrate the hydrophilic layer and the hydrophobic layer and directly connect to the reaction region, and the hydrophobic layer has cavities, thereby forming the microchannels between the substrate and the hydrophilic layer, thereby providing a chip-based reagent box.

[0007] In one embodiment, the aperture is hexagonal, has a reaction section and a contraction-shaped buffer section of equal width, the buffer section has a wide end and a narrow end, the wide end extends to one end of the reaction section, and the narrow end communicates with a cavity in the hydrophobic layer, thereby providing a chip-based reagent box that communicates with the microchannel.

[0008] In one embodiment, the hydrophilic layer further comprises an adsorption portion, and the hydrophilic layer is disposed on the micro-layer away from the reaction area. A chip-based reagent box is provided, which has a drain outlet at one end of the channel, and the adsorption part is provided on the hydrophilic layer at the drain outlet.

[0009] In one embodiment, a chip-based reagent box is provided, further comprising a case, wherein the main body and the adsorption part are enclosed and positioned in the case, the case has a window part that is directly connected to the reaction area above the receptor through the opening, and the case has an air hole on the drain port side, and the drain port communicates with the air hole in the case.

[0010] Accordingly, based on a chip-based reagent box, another aspect of the present invention provides a detection method, which includes a dripping step, a washing step, and a detection step. In the dripping step, a sample liquid is directly dripped onto the reaction area through the opening, and reacted with the receptor during a reaction time. In the washing step, after the reaction time, a washing liquid is dripped into the reaction area so that the volume exceeds the capacity of the reaction area. The sample liquid and the washing liquid permeate into the microchannel and are discharged by capillary action. In the detection step, after the washing step, a detection liquid is dripped into the reaction area to detect the sample.

[0011] In one embodiment, the method comprises the step of dropping the sample liquid into the reaction area in a volume of 10 to 15 microliters, and the reaction time is 1 to 5 minutes.

[0012] In one embodiment, the detection method includes, in the washing step, first dropping a first drop of washing liquid into the reaction area to wash the sample liquid, and then allowing the sample liquid and washing liquid to penetrate into the microchannel to perform the first washing, and then dropping a second drop of washing liquid into the reaction area to wash the sample liquid again, and allowing the sample liquid and washing liquid to penetrate into the microchannel again to perform the second washing.

[0013] In one embodiment, a testing method is provided in which, in the testing step, a first drop of test liquid is first dropped into the reaction area, and the remaining cleaning liquid and test liquid are allowed to permeate into the microchannel by capillary action, and then a second drop of test liquid is dropped into the reaction area and allowed to remain in the reaction area, thereby testing the sample.

[0014] Accordingly, based on a chip-based reagent box, another aspect of the present invention provides a detection method, which includes a dripping step, a washing step, and a detection step. In the dripping step, a sample liquid is directly dripped onto the reaction area through the opening, and reacted with the receptor during a reaction time. In the washing step, after the reaction time, a washing liquid is dripped into the reaction area so that the volume exceeds the capacity of the reaction area. The sample liquid and the washing liquid permeate into the microchannel by capillary action and are discharged. In the detection step, after the washing step, a detection liquid is dripped into the reaction area to detect the sample. The sample liquid and the washing liquid or the detection liquid flowing through the microchannel are quickly absorbed by the adsorption part and discharged through the microchannel.

[0015] Accordingly, based on the chip-based reagent box, another aspect of the present invention provides a detection method, which includes a dripping step, a washing step, and a detection step. In the dripping step, a sample liquid is dripped directly onto the reaction area through the opening, and reacts with the receptor during a reaction time. In the washing step, after the reaction time, a washing liquid is dripped onto the reaction area so that the volume exceeds the capacity of the reaction area, and the sample liquid and the washing liquid permeate into the microchannel and are discharged by capillary action. In the detection step, after the washing step, a test liquid is dripped onto the reaction area to detect the sample. The pore size of the pores is adjusted to control the flow of the sample liquid, washing liquid, or test liquid discharged through the microchannel. Control the speed. [Effects of the Invention]

[0016] As a result, the chip-based reagent box and its detection method of the present invention directly drip sample liquid into the reaction area, react with the receptor, and after the reaction, the sample liquid and cleaning liquid or detection liquid are permeated into the microchannel and discharged, so that detection can be completed without the need for external force driving. Not only that, by reacting the sample liquid with the receptor when dripping it into the reaction area, the detection time can be shortened, thereby achieving the effect of improving detection efficiency. [Brief explanation of the drawings]

[0017] [Figure 1] 1 is a perspective view of a chip-based reagent box for biological testing according to an embodiment of the present invention. FIG. [Figure 2] FIG. 2 is a perspective view of the main body of the embodiment of the present invention. [Figure 3] FIG. 2 is an exploded schematic view of the main body of the embodiment of the present invention. [Figure 4] FIG. 2 is an exploded top view of the main body of the embodiment of the present invention. [Figure 5] FIG. 2 is an exploded schematic view of the case according to the embodiment of the present invention. [Figure 6] FIG. 1 is a partial schematic cross-sectional view of a chip-based reagent box for biological testing according to an embodiment of the present invention. [Figure 7] 1 is a flowchart of an inspection method according to an embodiment of the present invention. [Figure 8A] 10 is a reference diagram showing a state in which a sample liquid is directly dropped into a reaction area of ​​a chip-based reagent box for biological testing according to an embodiment of the present invention. FIG. [Figure 8B] This is a reference diagram following FIG. 8A, showing the state in which a cleaning solution is dropped onto the reaction region after the reaction time, and is then allowed to permeate into the microchannel and be discharged. [Figure 8C] 8B, and is a reference diagram showing the state in which a second drop of the test liquid is dropped into the reaction region after the washing step, and the sample is tested. [Figure 8D] 10 is a reference diagram showing a state in which the case of a chip-based reagent box for biological testing according to an embodiment of the present invention is adjusted to a reduced pore size. DETAILED DESCRIPTION OF THE INVENTION

[0018] In the above description, those skilled in the art will appreciate that modifications, including additions and / or substitutions, may be made. It will be apparent that any such modifications will not depart from the scope and spirit of the present invention. In order not to obscure the present invention, certain details may be omitted, but the present invention has been described in a manner that will enable one skilled in the art to practice the present invention without undue experimentation or error.

[0019] Please refer to Figures 1 to 8D. The present invention provides a chip-based reagent box 100 for biological testing and a detection method thereof 200. As shown in Figures 1 to 6, the chip-based reagent box 100 for biological testing includes a body 10 including a substrate 20 and a flow-guiding layer 30, and in an embodiment, further includes an adsorption part 40 and a case 50.

[0020] A pair of electrodes 21 is provided on the substrate 20, and a part of the pair of electrodes 21 has a reaction area 22, and the reaction area 22 is coated with a receptor 23 that binds to the detection target. The detection target of the receptor 23 may be, but is not limited to, an antibody, an antigen, a nucleic acid, or a small molecule.

[0021] The flow conducting layer 30 is disposed on top of the substrate 20 to form the main body 10. The flow conducting layer 30 has openings 31, which are disposed on-site with the reaction region 22. The receptors 23 are exposed to the outside of the main body 10 through the openings 31, allowing the sample liquid S to drip directly onto the reaction region 22 and react with the receptors 23 (as shown in FIG. 8A). A microchannel 11 exists between the flow conducting layer 30 and the substrate 20, and this microchannel 11 communicates with the reaction region 22. The sample liquid S is, for example, blood, but the present invention is not limited to this.

[0022] 2 to 4 , the flow guide layer 30 in this embodiment includes a hydrophilic layer 32 and a hydrophobic layer 33. The hydrophobic layer 33 is located between the substrate 20 and the hydrophilic layer 32. The hydrophilic layer 32 has capillary action and can absorb water, while the hydrophobic layer 33 does not absorb water. The openings 31 penetrate the hydrophilic layer 32 and the hydrophobic layer 33 and directly connect to the reaction region 22. The hydrophobic layer 33 has cavities, so that microchannels 11 are formed between the substrate 20 and the hydrophilic layer 32.

[0023] 3 , the hydrophilic layer 32 in this embodiment has a first perforation 321 and a drain port 322 at one end of the microchannel 11 remote from the reaction region 22, and the hydrophobic layer 33 has a second perforation 331, whereby the first perforation 321 and the second perforation 331 overlap to form the opening 31. The hydrophobic layer 33 also has a third perforation 332 forming a cavity, and the second perforation 331 and the drain port 322 communicate with both ends of the third perforation 332.

[0024] As shown in Figure 4, the aperture 31 is hexagonal and includes a reaction section 311 and a buffer section 312. The reaction section 311 has a uniform width, and the buffer section 312 has a constricted shape. The buffer section 312 has a wide end 312a and a narrow end 312b. The wide end 312a extends to one end of the reaction section 311, and the narrow end 312b communicates with the microchannel 11 via the third perforation 332 in the hydrophobic layer 33. In this embodiment, the receptor 23 is located in the reaction section 311, and the buffer section 312 provides buffering and flow rate adjustment functions. The hexagonal shape of the aperture 31 is merely an example, and the present invention is not limited thereto. By making the openings 31 hexagonal and connecting them directly to the reaction area 22, the hydrophilic layer 32 does not cause capillary action within the openings 31, and the sample liquid S is separated by the hexagonal structure of the openings 31, so that the sample liquid S and the cleaning liquid W or the test liquid T do not overflow even when dropped into the reaction area 22.

[0025] 5 and 6, the main body 10 and the adsorbent part 40 are enclosed in a case 50 and positioned. The case 50 has a window 51 and is connected to the reaction region 22 on the receptor 23 through the opening 31, and the case 50 has an air hole 52 on the drain port 322 side. The drain port 322 is connected to the air hole 52 in the case 50. In one embodiment, the adsorbent part 40 is disposed on the hydrophilic layer 32 at the drain port 322.

[0026] As shown in FIGS. 5 and 6 , the case 50 has a fixing portion 53 corresponding to the suction portion 40. The fixing portion 53 holds the suction portion 40 in the drainage port 322, thereby positioning the suction portion 40 so that it does not displace. In this embodiment, the fixing portion 53 is two ribs protruding from the case 50. However, the fixing portion 53 is not limited to the above-described embodiment in which the fixing portion 53 is a rib. For example, the fixing portion 53 may be one or three or more ribs. The shape of the rib is linear, but is not limited thereto, and may be, for example, round or sheet-shaped. Simply put, any object that can achieve the above-described function of holding the suction portion 40 in the drainage port 322 is covered by the scope of protection of the fixing portion 53 of the present invention. Furthermore, the case 50 has a slot 54 corresponding to the main body 10, into which the main body 10 can be inserted, thereby allowing the reaction region 22 to be positioned within the window portion 51. The case 50 also has a rib portion 55 in the slot 54, and the main body 10 is held and positioned in the slot 54 by the rib portion 55. In this embodiment, the case 50 is formed by combining two half case portions 56 together.

[0027] As shown in FIG. 7, the inspection method 200 includes a dropping step 201 , a cleaning step 202 and an inspection step 203 .

[0028] In the dropping step 201, the sample liquid S is dropped directly into the reaction area 22 through the opening 31 (see FIG. 8A), and reacts with the receptor 23 during the reaction time. In one embodiment, the sample liquid S reacts in the reaction area 22 in a volume of 10 to 15 μL, and the reaction time is 1 to 5 minutes. The receptor 23 of the present invention is, but is not limited to, an immunoglobulin, a nucleic acid probe, a chemical molecule, or a functional protein. The sample liquid of the present invention contains a detection target capable of binding to the receptor. In one embodiment, the receptor 23 is an IgG antibody or an IgM antibody, and the detection target contained in the sample liquid S is a virus or microorganism capable of specifically binding to the IgG antibody or IgM antibody.

[0029] In one embodiment, the receptor 23 is an immunoglobulin, in particular an antibody to a coronavirus receptor binding domain protein (anti-COVID-19 RBD antibody), with a concentration of 1-10 μg / mL. The detection target contained in the sample liquid S is, in this embodiment, a coronavirus receptor binding domain protein (COVID-19 RBD) contained in saliva or nasal mucus, with a detection range of 1-1000 pg / mL.

[0030] In the washing step 202, after the reaction time, a washing solution W is dropped onto the sample solution S so that the volume exceeds the capacity of the reaction region 22. The sample solution S and the washing solution W are then permeated into the microchannel 11 by capillary action and discharged. In this embodiment, the capillary action is provided by the hydrophilic layer 32. In one embodiment, in the washing step 202 (see FIG. 8B ), a first drop of 35 μL of washing solution W is dropped onto the reaction region 22 to wash the sample solution S, thereby removing substances that do not specifically bind to the receptor 23 in the sample solution S, and the sample solution S and the washing solution W are permeated into the microchannel 11 to complete the first washing. Next, a second drop of 35 μL of washing solution W is dropped onto the reaction region 22 to wash the sample solution S again, thereby removing substances that do not specifically bind to the receptor 23 in the sample solution S, and the sample solution S and the washing solution W are permeated into the microchannel 11 again to complete the second washing. This prevents the non-specific binding substances remaining in the sample liquid S in the reaction area 22 from affecting the detection results.

[0031] In the detection step 203, after the cleaning step 202, the test liquid T is dropped into the reaction region 22 for sample detection. In one embodiment, in the detection step 203 of this embodiment, a first drop of 35 μL of the test liquid T is dropped into the reaction region 22, and the test liquid T can permeate into the microchannel 11 by capillary action (see FIG. 8B). Then, a second drop of the test liquid T is dropped into the reaction region 22, and the second drop of the test liquid T remains in the reaction region 22 (see FIG. 8C), and the sample is detected by an electrochemical reaction with the reacted receptor 23.

[0032] In this embodiment, the sample liquid S and the cleaning liquid W permeate the microchannel 11 by capillary action in the cleaning step 202, and the test liquid T permeates the microchannel 11 by capillary action in the testing step 203. These liquids fill the microchannel 11, and when they are discharged through the drain port 322, they are quickly absorbed by the adsorption unit 40. This can accelerate the discharge of the sample liquid S and the cleaning liquid W in the cleaning step 202 or the test liquid T in the testing step 203 from the microchannel 11. The provision of the adsorption unit 40 is merely a preferred embodiment, and the present invention is not limited thereto. That is, the sample liquid S and the cleaning liquid W or the test liquid T in the microchannel 11 can naturally be discharged through the drain port 322 even if the microchannel 11 is filled. The difference is that the presence of the adsorption unit 40 allows the sample liquid S, the cleaning liquid W, or the test liquid T to be quickly absorbed and accelerated to be discharged from the microchannel 11.

[0033] In one embodiment, the pore size of the pores 52 is adjusted to allow the sample liquid S and the washing liquid The speed at which the cleaning liquid W or the test liquid T is discharged from the microchannel 11 is controlled. As shown in FIGS. 8A to 8C, when the pores 52 of the case 50 are large, the exhaust capacity is large, and therefore the sample liquid S, cleaning liquid W, or test liquid T is discharged from the microchannel 11 more quickly. Furthermore, as shown in FIG. 8D, when the pores 52 of the case 50 are small, the exhaust capacity is small, and therefore the speed at which the sample liquid S, cleaning liquid W, or test liquid T is discharged from the microchannel 11 is slow. The control of the pore size of the pores 52 is determined based on the need for the speed at which the sample liquid S, cleaning liquid W, or test liquid T is discharged from the microchannel 11. In this embodiment, the pores 52 have a fixed pore size in the case 50, and the pore size of the pores 52 can be adjusted by replacing the case 50, but the present invention is not limited thereto. For example, a flow valve (not shown) can be installed in the case 50, and pores 52 can be formed in the flow valve, and the size of the pore diameter can be adjusted by the flow valve, and similarly, the speed at which the sample liquid S and the cleaning liquid W or the test liquid T are discharged from the microchannel 11 can be controlled.

[0034] According to the above description, the characteristics of the present invention are as follows:

[0035] (1) In the chip-based reagent box 100 for biological testing and the detection method 200 thereof according to the present invention, the sample liquid S is directly dropped into the reaction area 22, first reacting the sample liquid S with the receptor 23, and then the sample liquid S and the washing liquid W are allowed to permeate the microchannel 11 and be discharged. In contrast, in conventional detection devices, the sample liquid is first guided to the reaction area by capillary action and then reacted with the receptor. Therefore, the present invention can complete sample detection without requiring external driving, and can also directly react the sample liquid S with the receptor 23 when dropped into the reaction area 22. This avoids the delay in detection time caused by guiding the sample liquid to the reaction area, thereby shortening the detection time and thereby achieving the effect of efficiently improving detection efficiency.

[0036] (2) The openings 31 are hexagonal and directly communicate with the reaction region 22, so that capillary action does not occur in the hydrophilic layer 32 at the openings 31. The hexagonal openings 31 act as a barrier to the sample liquid S, preventing overflow of the sample liquid S and the cleaning liquid W or test liquid T when they are dropped into the reaction region 22. This helps to smooth the test process and further improve test efficiency.

[0037] (3) By providing the adsorption unit 40, when the sample liquid S and the cleaning liquid W or the test liquid T are discharged from the microchannel 11, they are immediately absorbed by the adsorption unit 40. This has the effect of accelerating the discharge compared to natural discharge, and ultimately helps to further improve the test efficiency.

[0038] Although the present invention has been described above with reference to preferred embodiments, the above embodiments are merely used to explain the present invention and should not be construed as limiting the scope of the present invention to those skilled in the art. It should be noted that any equivalent modifications or replacements of the above embodiments are also understood to be included within the scope of the present invention. Therefore, the scope of protection of the present invention is defined by the claims.

Claims

1. a body, the body comprising: a substrate provided with a pair of electrodes, the pair of electrodes including a reaction region, the reaction region being coated with a receptor that binds to the detection target; a flow guide layer provided on the substrate, the flow guide layer having openings disposed at the same positions as the reaction regions; The receptor is exposed to the outside of the body through the opening, so that a sample liquid can be directly dropped into the reaction area and react with the receptor; a microchannel communicating with the reaction region is present between the flow guide layer and the substrate; a washing solution is added to the sample solution after the reaction, and the washing solution is permeated into the microchannel and then discharged, so that both the sample solution and the washing solution are absorbed into the microchannel by capillary action; The flow-guiding layer includes a hydrophilic layer having capillary action and a hydrophobic layer that does not absorb water, the hydrophobic layer is disposed between the substrate and the hydrophilic layer; the openings penetrate the hydrophilic layer and the hydrophobic layer directly to the reaction regions; A chip-based reagent box for biological testing, wherein the hydrophobic layer has a cavity, thereby forming the microchannel between the base material and the hydrophilic layer.

2. The apertures are hexagonal and have equal width reaction sections and contraction buffer sections; the buffer stage having a wide end and a narrow end; the broad end extends to one end of the reaction stage; The chip-based reagent box according to claim 1 , wherein the narrow end communicates with the cavity of the hydrophobic layer, thereby communicating with the microchannel.

3. Further includes an adsorption portion, the hydrophilic layer has a drain outlet at one end of the microchannel remote from the reaction region; The chip-based reagent box according to claim 1 , wherein the adsorption portion is provided on the hydrophilic layer at the drainage port.

4. Also includes a case, the main body and the suction part are positioned by being enclosed in the case; the case has a window portion directly connected to the reaction area above the receptor through the opening, and the case has an air hole on one side of the drain hole; The chip-based reagent box according to claim 3 , wherein the drain hole communicates with the air hole in the case.

5. The chip-based reagent box according to claim 1 , wherein the receptor is selected from the group consisting of immunoglobulins, nucleic acid probes, chemical molecules, and functional proteins.

6. a dropping step of dropping the sample liquid directly into the reaction area through the opening and allowing it to react with the receptor during a reaction time; a washing step in which, after the reaction time, the washing solution is dropped into the reaction region so as to exceed the capacity of the reaction region, and the sample solution and the washing solution permeate into the microchannel by capillary action and are then discharged; 3. The method for testing a chip-based reagent box according to claim 1, further comprising: a testing step of testing the sample by dropping a test liquid into the reaction region after the washing step.

7. 7. The method according to claim 6, wherein in the dropping step, the reaction is carried out in the reaction region with the sample liquid in a volume of 10 to 15 microliters, and the reaction time is 1 to 5 minutes.

8. In the washing step, a first drop of the cleaning solution is dropped into the reaction region to clean the sample solution, and the sample solution and the cleaning solution are allowed to penetrate into the microchannel to perform a first cleaning; The detection method according to claim 7, wherein a second drop of the cleaning solution is then dropped into the reaction region to clean the sample liquid again, and the sample liquid and the cleaning solution are again allowed to penetrate into the microchannel to perform a second cleaning.

9. In the inspection step, a first drop of the test liquid is dropped into the reaction region, and the remaining cleaning liquid and test liquid are allowed to permeate into the microchannel by capillary action; 8. The method according to claim 7, further comprising the steps of: dropping a second drop of the test liquid into the reaction area; and allowing the second drop of the test liquid to remain in the reaction area, thereby testing the sample.

10. a dropping step of dropping the sample liquid directly into the reaction area through the opening and allowing it to react with the receptor during a reaction time; a washing step in which, after the reaction time, the washing solution is dropped into the reaction region so as to exceed the capacity of the reaction region, and the sample solution and the washing solution permeate into the microchannel by capillary action and are then discharged; a measuring step of measuring the sample by dropping a measuring liquid into the reaction area after the washing step; 4. The method for inspecting a chip-based reagent box according to claim 3, wherein the sample liquid and the cleaning liquid or the inspection liquid flowing through the microchannel is quickly absorbed by the adsorption part and discharged through the microchannel.

11. In the dropping step, the sample liquid is dropped in a volume of 10 to 15 microliters.

11. The method according to claim 10, wherein a reaction is carried out in the reaction region, and the reaction time is 1 to 5 minutes.

12. In the washing step, a first drop of the cleaning solution is dropped into the reaction region to clean the sample solution, and the sample solution and the cleaning solution are allowed to penetrate into the microchannel to perform a first cleaning; The detection method according to claim 11, wherein a second drop of the cleaning solution is then dropped into the reaction region to clean the sample liquid again, and the sample liquid and the cleaning solution are again allowed to penetrate into the microchannel to perform a second cleaning.

13. In the inspection step, a first drop of the test liquid is dropped into the reaction region, and the remaining cleaning liquid and test liquid are allowed to permeate into the microchannel by capillary action; The detection method according to claim 11, further comprising the steps of: dropping a second drop of the test liquid into the reaction area; and allowing the second drop of the test liquid to remain in the reaction area, thereby detecting the sample.

14. a dropping step of dropping the sample liquid directly into the reaction area through the opening and allowing it to react with the receptor during a reaction time; a washing step in which, after the reaction time, the washing solution is dropped into the reaction region so as to exceed the capacity of the reaction region, and the sample solution and the washing solution permeate into the microchannel by capillary action and are then discharged; a measuring step of dropping a measuring solution into the reaction area after the washing step to measure the sample; The method for testing a chip-based reagent box according to claim 4, wherein the flow rate of the sample liquid and cleaning liquid or the test liquid discharged through the microchannel is controlled by adjusting the pore size of the pores.

15. 15. The method according to claim 14, wherein in the dropping step, the reaction is carried out in the reaction area with the sample liquid in a volume of 10 to 15 microliters, and the reaction time is 1 to 5 minutes.

16. In the washing step, a first drop of the cleaning solution is dropped into the reaction region to clean the sample solution, and the sample solution and the cleaning solution are allowed to penetrate into the microchannel to perform a first cleaning; The detection method according to claim 15, wherein a second drop of the cleaning solution is then dropped into the reaction region to clean the sample liquid again, and the sample liquid and the cleaning solution are allowed to penetrate into the microchannel to perform a second cleaning.

17. In the inspection step, a first drop of the test liquid is dropped into the reaction region, and the remaining cleaning liquid and test liquid are allowed to permeate into the microchannel by capillary action; 16. The method according to claim 15, further comprising dropping a second drop of the test liquid into the reaction area, allowing the second drop of the test liquid to remain in the reaction area, and then testing the sample.

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