Micro-fluidic chip capable of realizing two reactions

Abstract

The utility model provides a micro-fluidic chip capable of realizing twice reactions, the micro-fluidic chip comprises a chip substrate, an upper cover plate and an extraction structural member, the upper cover plate and the extraction structural member are both located above the chip substrate, and the upper cover plate and the extraction structural member are arranged adjacently; an electrode array is arranged on the chip substrate; the electrode array comprises a first liquid separation electrode area, a first reaction electrode area and a second reaction electrode area which are sequentially arranged in the first direction, the first reaction electrode area comprises at least one first reaction electrode set, and the second reaction electrode area comprises at least one second reaction electrode set. The first reaction electrode groups and the second reaction electrode groups are the same in number and are correspondingly arranged; each of the first reaction electrode group and the second reaction electrode group comprises a uniformly-mixed electrode structure, each uniformly-mixed electrode structure comprises at least three electrode plates, and at least two electrode plates are arranged along the circumferential direction of the uniformly-mixed electrode structure. The micro-fluidic chip disclosed by the utility model can be used for carrying out multi-step reaction, and is high in detection efficiency and good in uniform mixing effect.

Description

Technical Field

[0001] The utility model relates to the technical field of microfluidics, in particular to a microfluidic chip capable of realizing two reactions. Background Art

[0002] A microfluidic chip integrates operation units such as sample preparation, reaction, and detection in the analysis processes of chemistry, biology, medicine, etc. onto a carrier at the micron scale, and automatically completes the whole analysis process.

[0003] Most of the existing microfluidic chips can only perform one-step reactions. After the reaction is completed, the product is taken out and then the next reaction is carried out. Since in these multi-step reactions, the opening of the product is involved, it is easy to cause aerosol pollution in the laboratory. At the same time, the transfer of the product, especially the transfer to multiple reaction vessels for multi-target reactions, will make manual operations very complicated.

[0004] In addition, most of the existing microfluidic chips are used for isothermal amplification reactions. Once a temperature-variable amplification reaction is involved, bubbles are easily generated during the high-temperature heating process. These bubbles pose a great risk to subsequent liquid movement and fluorescence data collection. At the same time, the mixing effect inside the liquid during the reaction process is not ideal, which has a great impact on the reaction efficiency and quality. Summary of the Utility Model

[0005] The purpose of the utility model is to provide a microfluidic chip capable of realizing two reactions, which is used to solve the problems that the existing microfluidic chips cannot perform multi-step reactions, the liquid inside is not evenly mixed, and bubbles are easily generated during the high-temperature heating process, affecting the use.

[0006] To achieve the purpose of the utility model, the microfluidic chip capable of realizing two reactions of the utility model includes a chip substrate, an upper cover plate, and an extraction structural member. The upper cover plate and the extraction structural member are both located above the chip substrate, and the upper cover plate and the extraction structural member are arranged adjacent to each other; a liquid activity cavity is formed between the chip substrate, the upper cover plate, and the extraction structural member. An electrode array is arranged on the chip substrate, and the liquid activity cavity is correspondingly arranged with the electrode array; the electrode array includes a first liquid separation electrode area, a first reaction electrode area, and a second reaction electrode area arranged in sequence along a first direction. The first reaction electrode area includes at least one first reaction electrode group, and the second reaction electrode area includes at least one second reaction electrode group. The number of the first reaction electrode groups is the same as that of the second reaction electrode groups, and the first reaction electrode groups and the second reaction electrode groups are arranged in one-to-one correspondence; both the first reaction electrode group and the second reaction electrode group include at least one mixing electrode structure, and each mixing electrode structure includes at least three electrode sheets, and at least two of the electrode sheets are arranged along the circumferential direction of the mixing electrode structure. Preferably, the first electrode sheet is located at the center of the mixing electrode structure, and the other electrode sheets are arranged along the circumferential direction of the first electrode sheet.

[0007] As can be seen from the above solution, the microfluidic chip of the present utility model can perform two fully automated reactions. Only by adding a sample, the sample can be input and the result can be output. The nucleic acid extraction process and the two reaction processes can be automatically completed within the microfluidic chip, saving the cumbersome steps of manual extraction and manual transfer of products, and at the same time preventing the laboratory from being contaminated by the products. In addition, the hybrid electrode structure of the present utility model can play a role in stirring the liquid, thereby increasing the convection of each component inside the liquid. While greatly improving the mixing effect of the liquid, it also makes the temperature inside the liquid more uniform during the reaction process. At the same time, since the liquid moves around the center of the hybrid electrode structure, most of the bubbles generated during the reaction heating process are pushed away from the hybrid electrode structure during the repeated movement of the liquid and stay in the nearby area, and the remaining bubbles will also be pushed to the edge of the hybrid electrode structure during the movement of the liquid, without affecting the movement of the liquid and the acquisition of fluorescence data.

[0008] A further solution is that a second liquid separation electrode area is also provided between the first reaction electrode area and the second reaction electrode area. The second liquid separation electrode area is adjacent to the first reaction electrode area and the second reaction electrode area respectively. The second liquid separation electrode area includes at least one second liquid separation electrode group. The number of the second liquid separation electrode groups is the same as that of the second reaction electrode groups, and the second liquid separation electrode groups and the second reaction electrode groups are arranged in one-to-one correspondence; the second reaction electrode group includes at least two reaction points and a connecting electrode. The connecting electrode is adjacent to the second liquid separation electrode group and the reaction point respectively, and the reaction point is a mixing electrode structure.

[0009] As can be seen from the above solution, the setting of the second liquid separation electrode area and the reaction points enables the microfluidic chip to not only perform multi-target reactions and detections simultaneously, but also perform different detection items at different reaction points, making the detection efficiency higher and more flexible.

[0010] A further solution is that the microfluidic chip further includes a buffer pre-storage area. The buffer pre-storage area is located above the chip substrate, and the buffer pre-storage area and the extraction structural member are arranged adjacent to each other along the second direction, and the second direction is perpendicular to the first direction; the electrode array further includes a third liquid separation electrode area. The third liquid separation electrode area is arranged along the second direction with the first liquid separation electrode area. One side of the third liquid separation electrode area is adjacent to the first reaction electrode area, and the other side of the third liquid separation electrode area is communicated with the buffer pre-storage area.

[0011] As can be seen from the above solution, the structure related to the buffer plays a role in diluting the product of the first reaction, ensuring that the amount of the diluted product is sufficient for liquid separation for the second reaction.

[0012] A further solution is that the first reaction electrode group includes a first mixing electrode group and a second mixing electrode group arranged in sequence along the first direction, and both the first mixing electrode group and the second mixing electrode group are mixing electrode structures.

[0013] As can be seen from the above solution, the setting of the first mixing electrode group and the second mixing electrode group makes the liquid mixing effect better, which is suitable for the microfluidic chip with the reaction solution added on-site, and is more suitable for multi-step reactions.

[0014] A further solution is that the microfluidic chip further includes a second structural member, the second structural member is located above the chip substrate, and the second structural member and the extraction structural member are arranged adjacent to each other in the second direction. The second structural member includes a reaction solution sampling port and a reaction solution storage area, and the reaction solution sampling port is communicated with the reaction solution storage area; the electrode array further includes a fourth liquid separation electrode area, the fourth liquid separation electrode area and the first liquid separation electrode area are arranged in the second direction, one side of the fourth liquid separation electrode area is adjacent to the first reaction electrode area, and the other side of the fourth liquid separation electrode area is communicated with the reaction solution storage area.

[0015] As can be seen from the above solution, the setting of the reaction solution sampling-related structure enables the reaction system that cannot be dried on the chip to be added to the chip in the way of adding on-site as needed, making the new application range of the microfluidic wider and giving users more room for choice.

[0016] A further solution is that a hydrophobic coating is provided at the position of the upper cover plate facing the electrode array and not corresponding to the electrode array.

[0017] As can be seen from the above solution, the above modification of the upper cover plate makes the liquid firmly adsorbed at the reaction position, thereby preventing the bubbles generated by high temperature from squeezing away the liquid. At the same time, the liquid adsorbed by each reaction is quantified and will not affect the liquid transfer.

[0018] A further solution is that a pre-stored drying reagent is fixed on the first reaction electrode area and / or the second reaction electrode area.

[0019] As can be seen from the above solution, the reagent of the reaction solution can be fixed on the chip in advance by drying, saving chip space and being convenient to use.

[0020] A further solution is that the microfluidic chip further includes a housing, the chip substrate and the upper cover plate are both located inside the housing, and the housing includes a housing bottom plate and a housing upper cover; the chip substrate is arranged on the housing bottom plate, and the housing upper cover is provided with a first through hole and a second through hole. The first through hole is correspondingly arranged with the extraction structural member, and the second through hole is located directly above the second reaction electrode area.

[0021] As can be seen from the above solution, the housing plays a good role in fixing, sealing and protecting other structures of the microfluidic chip, and the setting of the second through hole on the housing upper cover facilitates observing the detection results.

[0022] A further solution is that the liquid separation electrode region includes a liquid storage electrode part, a liquid separation electrode part, and a solid-liquid electrode part that are sequentially and adjacently arranged. The widths of both the liquid storage electrode part and the solid-liquid electrode part are greater than the width of the liquid separation electrode part; the extraction structural member includes a sample addition port, a lysis chamber, a washing chamber, and an elution chamber that are sequentially connected. The lysis chamber, the washing chamber, and the elution chamber are separated by paraffin valves. Magnetic beads are pre-stored at the sample addition port, and lysis solution is pre-stored in the lysis chamber.

[0023] As can be seen from the above solution, the above structure can accurately control the size of droplet splitting, and the size of the droplet can be determined by the size of the solid-liquid electrode part. The extraction structural member can quickly extract nucleic acids and improve the detection efficiency. Description of the Drawings

[0024] Figure 1 It is a structural diagram of the microfluidic chip of the present utility model.

[0025] Figure 2 It is an exploded view of the structure of the microfluidic chip of the present utility model.

[0026] Figure 3 It is a structural diagram of the electrode array and the chip substrate of the present utility model.

[0027] Figure 4 It is a structural diagram of the first liquid separation electrode region of the present utility model.

[0028] Figure 5 It is a structural diagram of the first mixing electrode group of the present utility model.

[0029] Figure 6 It is a structural diagram of the second mixing electrode group of the present utility model.

[0030] Figure 7 It is a structural diagram of the second reaction electrode group of the present utility model.

[0031] Figure 8 It is a bottom view of the extraction structural member of the present utility model.

[0032] Figure 9 It is a bottom view of the second structural member of the present utility model.

[0033] The present utility model will be further described below with reference to the drawings and embodiments. Detailed Embodiments

[0034] See Figures 1 to 9, the microfluidic chip capable of realizing two reactions in this embodiment includes a chip substrate 1, an upper cover plate 2, an extraction structural member 3, a second structural member 4, and a housing 5. The upper cover plate 2, the extraction structural member 3, and the second structural member 4 are all located above the chip substrate 1. The upper cover plate 2 and the extraction structural member 3 are arranged adjacent to each other along a first direction, and the second structural member 4 and the extraction structural member 3 are arranged adjacent to each other along a second direction, and the second direction is perpendicular to the first direction. The housing 5 includes a housing upper cover 51 and a housing bottom plate 52. The chip substrate 1 and the upper cover plate 2 are both located inside the housing. The chip substrate 2 is arranged on the housing bottom plate 52. A protrusion 521 is provided on the side of the housing bottom plate 52 facing the chip substrate 1. A liquid activity cavity is formed between the protrusion 521, the chip substrate 1, the upper cover plate 2, the extraction structural member 3, and the second structural member 4.

[0035] See Figure 3 , an electrode array 6 is provided on the chip substrate 1, and the liquid activity cavity is correspondingly arranged with the electrode array 6. The electrode array 6 includes a first liquid separation electrode area 61, a first reaction electrode area 62, a second liquid separation electrode area 63, and a second reaction electrode area 64 arranged in sequence along the first direction. The first reaction electrode area 62 includes at least one first reaction electrode group 621. The second liquid separation electrode area 63 includes at least one second liquid separation electrode group 631. The second reaction electrode 64 includes at least one second reaction electrode group 641. The number of the first reaction electrode groups 621, the number of the second liquid separation electrode groups 631, and the number of the second reaction electrode groups 641 are the same, and the first reaction electrode groups 621, the second liquid separation electrode groups 631, and the second reaction electrode groups 641 are arranged in one-to-one correspondence. In this embodiment, the number of the first reaction electrode groups 621, the second liquid separation electrode groups 631, and the second reaction electrode groups 641 is 8 each.

[0036] The electrolysis array 6 further includes a third liquid separation electrode area 65 and a fourth liquid separation electrode area 66. The first liquid separation electrode area 61, the third liquid separation electrode area 65, and the fourth liquid separation electrode area 66 are arranged along the second direction. One side of the third liquid separation electrode area 65 is adjacent to the first reaction electrode area 62, and one side of the fourth liquid separation electrode area 66 is adjacent to the first reaction electrode area 62.

[0037] A heating wire may or may not be provided below the reaction electrode area and the liquid separation electrode area, and the microfluidic chip is heated by a heating device on the microfluidic detection device.

[0038] Among them, the first liquid separation electrode area 61, the second liquid separation electrode area 63, the third liquid separation electrode area 65, and the fourth liquid separation electrode area 66 are all liquid separation electrode areas. See Figure 4, wherein the first liquid separation electrode region 61 includes a liquid storage electrode portion 671, a liquid separation electrode portion 672, and a solid-liquid electrode portion 673 that are sequentially adjacent to each other along the first direction. The widths of both the liquid storage electrode portion 671 and the solid-liquid electrode portion 673 are greater than the width of the liquid separation electrode portion 672. The number and shape of the electrode sheets of the liquid storage electrode portion 671, the liquid separation electrode portion 672, and the solid-liquid electrode portion 673 can be determined according to design requirements. The liquid separation electrode portion 672 includes a liquid separation electrode 6721 and a liquid transfer electrode 6722. The liquid transfer electrode 6722 is located between the liquid separation electrode 6721 and the liquid storage electrode portion 671. In this embodiment, the number of the liquid separation electrodes 6721 is one, and the shape is approximately arrow-shaped. The widths of both ends of the liquid separation electrode 6721 adjacent to the liquid storage electrode portion 671 and the solid-liquid electrode portion 673 are smaller than the widths of the liquid storage electrode portion 671 and the solid-liquid electrode portion 673. The microfluidic chip precisely splits droplets through the liquid separation electrode region, and the size of the split daughter droplets is determined by the size of the solid-liquid electrode region 673. The structures of other liquid separation electrode regions can be the same as that of the first liquid separation electrode region 61, or can be as shown in the related structure in the Chinese patent with the publication number CN218393734U.

[0039] See Figures 5 to 7 , the first reaction electrode group 621 includes a first mixing electrode group 6211 and a second mixing electrode group 6212 that are arranged in sequence along the first direction. See Figure 7 , the second reaction electrode group 641 includes at least two reaction points 6411 and a connecting electrode 6412. A pre-stored drying reagent is fixed on the reaction point 6411. In this embodiment, one second reaction electrode group 641 contains six reaction points 6411, and the connecting electrode 6412 is adjacent to the second liquid separation electrode group 631 and the reaction point 6411 respectively. The reaction point 6411, the first mixing electrode group 6211, and the second mixing electrode group 6212 are all mixing electrode structures. The mixing electrode structure includes at least three electrode sheets, and at least two of the electrode sheets are arranged along the circumferential direction of the mixing electrode structure.

[0040] Specifically, see Figure 5 , one first mixing electrode group 6211 includes five electrode sheets. Among them, the first electrode sheet 601 is located at the center of the first mixing electrode group 6211, and the second electrode sheet 602, the third electrode sheet 603, the fourth electrode sheet 604, and the fifth electrode sheet 605 are arranged along the circumferential direction of the first electrode sheet 601. See Figure 6 , one second mixing electrode group 6212 includes six electrode sheets. Among them, the first electrode sheet 681 is located at the center of the second mixing electrode group 6212, and the second electrode sheet 682, the third electrode sheet 683, the fourth electrode sheet 684, and the fifth electrode sheet 685 are arranged along the circumferential direction of the first electrode sheet 681. The sixth electrode sheet 686 is arranged at one end of the fifth electrode sheet 685 away from the first mixing electrode group 6211. See Figure 7, a reaction point 6411 includes three electrode plates. Among them, the first electrode plate 691 is located at the center of the reaction point 6411, and the second electrode plate 692 and the third electrode plate 693 are arranged along the circumferential direction of the first electrode plate 691. The shape of the electrode plate can be designed according to requirements without specific limitations.

[0041] The upper cover plate 2 is modified. Specifically, a hydrophobic coating (not shown in the figure) is provided at the position where the upper cover plate 2 faces the electrode array 6 and does not correspond to the electrode array 6, and there are areas where the hydrophobic coating is not provided in other places where the upper cover plate 2 faces the electrode array 6. The specific method of coating the coating can be to first coat a hydrophobic layer on the upper cover plate 2, and then wipe off the hydrophobic layer on the surface of the area where the hydrophobic coating is not required. The specific methods of setting and wiping off the hydrophobic layer are well-known methods to those skilled in the art.

[0042] See Figure 8 , the extraction structure 3 includes a sample addition port 31, a lysis chamber 32, a washing chamber 33, and an elution chamber 34 that are connected in sequence. In this embodiment, the washing chamber 33 includes a first washing chamber 331 and a second washing chamber 332. The lysis chamber 32, the washing chamber 33, and the elution chamber 34 are separated by paraffin valves. Magnetic beads are pre-stored at the sample addition port 31, and lysis solution is pre-stored in the lysis chamber 32. The specific structure and usage method of the extraction structure 3 can refer to the content disclosed in the Chinese patent with the publication number CN219886036U.

[0043] See Figure 9 , the second structure 4 includes a buffer pre-storage area 41 and a reaction solution storage area 42 arranged along the second direction. A reaction solution addition port 43 is provided above the reaction solution storage area 42, and the reaction solution addition port 43 is communicated with the reaction solution storage area 42. The buffer pre-storage area 41 is communicated with the third liquid separation electrode area 65, and the reaction solution storage area 42 is communicated with the fourth liquid separation electrode area 66. In some embodiments, the reaction solution storage area 42, the fourth liquid separation electrode area 66, and the first mixing electrode group 6211 may not be included, and the reaction solution can be pre-dried in the second mixing electrode group 6212.

[0044] A first through hole 511, a second through hole 512, and a third through hole 513 are provided on the outer shell upper cover 51. The first through hole 511 is correspondingly arranged with the extraction structure 3, and the sample addition port 31 of the extraction structure 3 passes through the first through hole 511 to fix the position of the extraction structure 3. The second through hole 512 is directly above the second reaction electrode area 64. The upper cover plate 2 is made of a transparent material, and the second through hole 512 is used to observe the detection result. The third through hole 513 is correspondingly arranged with the second structure 4, and the reaction solution addition port 43 passes through the third through hole 513 to fix the position of the second structure 4. The outer shell bottom plate 52 is provided with a fourth through hole 522, and the setting of the fourth through hole 522 facilitates the contact between the chip substrate 1 and the heating device on the microfluidic detection device for heating.

[0045] The usage method of the microfluidic chip capable of realizing two reactions in this embodiment specifically includes the following steps;

[0046] S1 Nucleic acid extraction: Add the sample solution into the sample inlet 31 of the extraction structural member 3. The sample solution with magnetic beads sequentially passes through the lysis chamber 32, the first washing chamber 331, the second washing chamber 332, and the elution chamber 3 / 4 for washing and eluting nucleic acids to obtain a nucleic acid solution. The specific working method of the extraction structural member 3 can refer to the working method disclosed in the Chinese patent with the publication number CN219886036U;

[0047] S2 Nucleic acid liquid separation: The nucleic acid solution is separated by the action of the first liquid separation electrode region 61 to form sub-droplets;

[0048] The control method of the first liquid separation electrode region 61, that is, the liquid separation method, is as follows: The nucleic acid solution covers the liquid storage electrode part 671. At this time, the pipetting electrode 6722, the liquid separation electrode 6721, and the solid-liquid electrode part 673 are energized, and the nucleic acid solution moves to the solid-liquid electrode part 673, so that the nucleic acid solution simultaneously wets the pipetting electrode 6722, the liquid separation electrode 6721, the solid-liquid electrode part 673, and the liquid storage electrode part 671. Then the solid-liquid electrode part 673, the pipetting electrode 6722, and the liquid storage electrode part 671 are energized, and the liquid separation electrode 6721 is not energized to cause the liquid droplet to break, so that the sub-droplets enter different first mixing electrode groups 6211. The specific liquid separation method can refer to that disclosed in CN218393734U;

[0049] S3 First reaction: The sub-droplets enter the first reaction electrode region 62 and are mixed and reacted with the reaction solution there;

[0050] The first reaction in this embodiment is a pcr amplification reaction. The specific control method of the first reaction electrode region 62 is as follows: The sub-droplets enter the first mixing electrode group 6211. The reaction solution of the first reaction is added into the microfluidic chip through the reaction solution sample inlet 43 and is separated by the fourth liquid separation electrode region 66 and enters different first mixing electrode groups 6211 respectively. The liquid separation method of the fourth liquid separation electrode region 66 here is the same as that of the first liquid separation electrode region 61.

[0051] The sub-droplets are mixed with the reaction solution. The control method of the first mixing electrode group 6211, that is, the specific mixing method, is as follows: First, energize the first electrode plate 601, the second electrode plate 602, and the third electrode plate 603; then energize the first electrode plate 601, the third electrode plate 603, and the fourth electrode plate 604; then energize the first electrode plate 601, the fourth electrode plate 604, and the fifth electrode plate 605; then energize the first electrode plate 601, the fifth electrode plate 605, and the second electrode plate 602. Repeat the above steps multiple times to make the sub-droplets perform rotational motion with the reaction solution until they are mixed;

[0052] After the amplified reaction system solution after mixing is dropped, it moves to the second mixing electrode group 6212 under the action of energizing the electrode. PCR requires high-temperature amplification. The upper cover plate 2 corresponding to the first electrode plate 681 in the second mixing electrode group 6212 here has been modified accordingly, without a hydrophobic coating, so that the liquid is firmly adsorbed at the position of the first electrode plate 681, thereby preventing the bubbles generated by high temperature from squeezing away the liquid.

[0053] The reacted liquid is mixed with the buffer solution. The buffer solution is pre-stored in the buffer solution pre-storage area 41 and is separated under the action of energizing the electrode and the third liquid separation electrode area 65, and enters different second mixing electrode groups 6212 respectively. The liquid separation method of the third liquid separation electrode area 65 here is the same as that of the first liquid separation electrode area 61.

[0054] The reacted liquid is mixed with the buffer solution. The control method of the second mixing electrode group 6212, that is, the specific mixing method is: first, energize the first electrode plate 681, the fifth electrode plate 685, and the sixth electrode plate 686; then energize the first electrode plate 681, the second electrode plate 682, and the third electrode plate 683; then energize the first electrode plate 681, the third electrode plate 683, and the fourth electrode plate 684. Repeat the above steps multiple times to make the reacted liquid and the buffer solution perform rotational movement until they are mixed evenly. This mixing operation can also squeeze the large bubbles generated by high-temperature amplification to the edge of the electrode or outside the electrode area, preventing the large bubbles from affecting subsequent liquid transfer;

[0055] S4 Second reaction: After the first reaction is completed, the sub-droplets continue to move into the second reaction electrode area and are mixed and reacted with the drying reagent there, and the fluorescence value of the reaction point is read.

[0056] The second reaction in this embodiment is a crisper reaction, and a drying reagent is pre-stored on each reaction point 6411. The diluted product after mixing enters different reaction points 6411 of the corresponding second reaction electrode group 641 through each second liquid separation electrode group 631. The liquid separation method of the second liquid separation electrode area 63 here is the same as that of the first liquid separation electrode area 61.

[0057] The control method of the reaction point 6411, that is, the mixing method is specifically: first, energize the first electrode plate 691 and the second electrode plate 692, and then energize the first electrode plate 691 and the third electrode plate 693. Repeat the above steps in a cycle to dry the diluted product solution on the drying reagent on each reaction point 6411 and mix it evenly. Subsequently, the detection results of each reaction point 6411 are read.

[0058] In this embodiment, 48 targets can be detected at one time to achieve 48-plex detection. Since the CRISPR method usually only uses the fluorescence of the FAM channel, if it is replaced with other detection methods that can use four fluorescence channels of FAM, HEX, ROX, and Cy5, up to 192-plex detection can be achieved, with high detection efficiency. At the same time, the microfluidic chip of this embodiment can perform two fully automated reactions. Only by adding the sample can the sample be input and the result be output. The nucleic acid extraction process and the two reaction processes can be automatically completed within the microfluidic chip, saving the cumbersome steps of manual extraction and manual transfer of products, and preventing the laboratory from being contaminated by the products. In addition, the hybrid electrode structure of the present invention can play a role in stirring the liquid, thereby increasing the convection of each component inside the liquid. While greatly improving the mixing effect of the liquid, it also makes the temperature inside the liquid more uniform during the reaction process. At the same time, since the liquid moves around the center of the hybrid electrode structure, most of the bubbles generated during the reaction heating process are pushed away from the hybrid electrode structure and stay in the nearby area during the repeated movement of the liquid. The remaining bubbles will also be pushed to the edge of the hybrid electrode structure during the movement of the liquid, without affecting the liquid movement and the acquisition of fluorescence data. The reaction solution can be dried or added immediately as needed, meeting the diverse design requirements of customers, having a wider application range, and being more convenient to use.

[0059] Finally, it should be emphasized that the above are only the preferred embodiments of the present invention and are not used to limit the present invention. For those skilled in the art, the present invention can have various changes and modifications. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A microfluidic chip capable of realizing double reactions, comprising a chip substrate, an upper cover plate and an extraction structure, wherein both the upper cover plate and the extraction structure are located above the chip substrate, and the upper cover plate and the extraction structure are arranged adjacent to each other; A liquid activity cavity is formed between the chip substrate, the upper cover plate and the extraction structure, an electrode array is arranged on the chip substrate, and the liquid activity cavity is arranged corresponding to the electrode array; Features: The electrode array comprises a first liquid-separating electrode area, a first reaction electrode area, and a second reaction electrode area sequentially arranged along a first direction, the first reaction electrode area comprises at least one first reaction electrode group, the second reaction electrode area comprises at least one second reaction electrode group, the number of the first reaction electrode groups is the same as the number of the second reaction electrode groups, and the first reaction electrode groups and the second reaction electrode groups are arranged in one-to-one correspondence; The first reaction electrode group and the second reaction electrode group each include at least one mixing electrode structure, and each of the mixing electrode structures includes at least three electrode sheets, wherein at least two of the electrode sheets are arranged along the circumference of the mixing electrode structure.

2. A microfluidic chip capable of realizing two reactions as claimed in claim 1, characterized in that: A second liquid-separating electrode area is further arranged between the first reaction electrode area and the second reaction electrode area, the second liquid-separating electrode area is adjacent to the first reaction electrode area and the second reaction electrode area respectively, the second liquid-separating electrode area includes at least one second liquid-separating electrode group, the number of the second liquid-separating electrode groups is the same as the number of the second reaction electrode groups, and the second liquid-separating electrode groups are arranged in one-to-one correspondence with the second reaction electrode groups; The second reaction electrode group includes at least two reaction points and a connecting electrode. The connecting electrode is adjacent to the second liquid-separating electrode group and the reaction point, respectively. The reaction point is the mixing electrode structure.

3. A microfluidic chip capable of realizing two reactions as claimed in claim 1, characterized in that: The microfluidic chip further includes a buffer pre-storage area, the buffer pre-storage area is located above the chip substrate, and the buffer pre-storage area is adjacent to the extraction structure along a second direction, and the second direction is perpendicular to the first direction; The electrode array also includes a third liquid-separating electrode area, which is arranged along the second direction with the first liquid-separating electrode area, one side of the third liquid-separating electrode area is adjacent to the first reaction electrode area, and the other side of the third liquid-separating electrode area is connected to the buffer pre-storage area.

4. A microfluidic chip capable of realizing double reactions as claimed in any one of claims 1 to 3, characterized in that: The first reaction electrode group includes a first mixing electrode group and a second mixing electrode group arranged in sequence along a first direction, and both the first mixing electrode group and the second mixing electrode group are the mixing electrode structures.

5. A microfluidic chip capable of realizing double reactions as claimed in any one of claims 1 to 3, characterized in that: The microfluidic chip further comprises a second structural member, the second structural member is located above the chip substrate, and the second structural member and the extraction structural member are arranged adjacent to each other along a second direction, the second structural member comprises a reaction liquid injection port and a reaction liquid storage area, and the reaction liquid injection port is connected to the reaction liquid storage area; The electrode array also includes a fourth liquid-separating electrode area, which is arranged along the second direction with the first liquid-separating electrode area, one side of the fourth liquid-separating electrode area is adjacent to the first reaction electrode area, and the other side of the fourth liquid-separating electrode area is connected to the reaction liquid storage area.

6. A microfluidic chip capable of realizing double reactions as claimed in any one of claims 1 to 3, characterized in that: A hydrophobic coating is disposed at a position of the upper cover plate facing the electrode array and not corresponding to the electrode array.

7. A microfluidic chip capable of realizing double reactions as claimed in any one of claims 1 to 3, characterized in that: A pre-stored drying reagent is fixed on the first reaction electrode area and / or the second reaction electrode area.

8. A microfluidic chip capable of realizing double reactions as claimed in any one of claims 1 to 3, characterized in that: The microfluidic chip further comprises a housing, the chip substrate and the upper cover are both located in the housing, and the housing comprises a housing bottom plate and a housing upper cover; The chip substrate is arranged on the housing bottom plate, and the housing upper cover is provided with a first through hole and a second through hole, wherein the first through hole is arranged corresponding to the extraction structure, and the second through hole is located directly above the second reaction electrode area.

9. A microfluidic chip capable of realizing double reactions as claimed in any one of claims 1 to 3, characterized in that: The liquid-separating electrode area comprises a liquid storage electrode portion, a liquid-separating electrode portion and a solid-liquid electrode portion which are arranged adjacent to each other in sequence, and the width of the liquid storage electrode portion and the width of the solid-liquid electrode portion are both greater than the width of the liquid-separating electrode portion; The extraction structure comprises a sample addition port, a lysis chamber, a cleaning chamber and an elution chamber which are connected in sequence. The lysis chamber, the cleaning chamber and the elution chamber are separated by a paraffin valve. Magnetic beads are pre-stored at the sample addition port, and lysis solution is pre-stored in the lysis chamber.

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