Full-liquid-phase centrifugal microfluidic detection chip for executing multi-step reaction

Abstract

The utility model discloses a full-liquid-phase centrifugal microfluidic detection chip for executing multi-step reaction, which comprises a main chip, the main chip is divided into a front side, a back side, a bottom shell, a preset box and a kit, the detection chip adopts a structure combining the main chip and the bottom shell, the back side of the main chip is provided with spikes, the bottom shell is provided with a diluent tank and a reaction reagent tank, and the diluent tank and the reaction reagent tank are arranged in the preset box. The liquid-phase reaction device is used for placing a preset box and a plurality of kits, and can realize multi-step full-liquid-phase reaction, so that the production and transportation are more convenient, the detection is more convenient, and the detected data are more accurate.

Description

Technical Field

[0001] The present invention belongs to the field of POCT biochemical immunoassay, and particularly relates to a fully liquid-phase centrifugal microfluidic detection chip that performs multi-step reactions. Background Art

[0002] A centrifugal microfluidic chip is a miniature analysis system that integrates sample liquid separation, quantification, centrifugation, reaction, and detection. Compared with traditional laboratory analysis, the microfluidic integrated chip simplifies the inspection process. Analysts only need to add samples to the microfluidic chip and place it in the supporting POCT inspection instrument. During this period, no further manual intervention is required, and the instrument automatically issues corresponding test results. Therefore, operators do not need professional experimental skill training, which improves work efficiency. Because the supporting instrument does not have a conventional cleaning pipeline system and pipetting system, its volume structure is light and small, and it does not require maintenance, making it suitable for use in primary medical care settings.

[0003] Existing centrifugal microfluidic chips are mainly used for biochemical detection except for molecular diagnostic experiments. Due to the limitation of production process level, they are generally limited to the use of solid freeze-dried reagents and simplified single-step reaction processes. The sample liquid and diluent are proportioned through various grooves and multiple microchannels on the chip, and the proportioned liquid enters the reaction tank through the microchannel to react with the freeze-dried reagent to obtain the required test results. However, this technology lacks the detection and calibration reference of the quality control liquid, making it difficult for users to timely detect problems with the instrument and reagents. When multi-step reaction detection is required, only multiple reaction reagents can be preset in the same reaction chamber, turning what should be multi-step reactions into one-step reactions, which will cause large deviations in the final data of multi-step reactions, and the final test results deviate too much to lose the reference value.

[0004] This chip has a small structure, is convenient to carry and operate, and can obtain test results quickly. However, limited by the freeze-dried reagent, there are problems such as complex production process, inconvenient transportation, and large deviation of test results. Therefore, it is very necessary to design a detection chip that does not require freeze-dried reagents. Summary of the Invention

[0005] (I) Technical problems to be solved.

[0006] In view of the deficiencies of the prior art, the present utility model provides a fully liquid-phase centrifugal microfluidic detection chip that performs multi-step reactions, and solves the problem of the complexity of freeze-dried reagents in the production and transportation processes mentioned in the above background art.

[0007] (II) Technical solutions.

[0008] To achieve the above object, the present utility model is realized by the following technical solutions: An all-liquid-phase centrifugal microfluidic detection chip for performing multi-step reactions, comprising a main chip, the main chip being divided into a front side and a back side. The front side of the main chip includes a sample loading port, a sample separation and quantification chamber, a diluent quantification chamber, a mixing chamber, a reaction chamber, a plurality of exhaust holes, a mixed liquid in-place detection hole, a sample in-place detection hole, a diluent loading port, a mixed liquid quantification chamber, a reaction reagent inlet, a colorimetric hole, and multiple groups of microchannels and multiple groups of pipes for connecting and controlling fluid flow. A sealing film is provided on the front side of the main chip. It is characterized in that: The back side of the main chip includes a pre-filled cartridge puncture tip, a reagent kit puncture tip, an expansion nail, a pre-filled cartridge upper mounting groove, a reagent kit upper mounting groove. The above detection chip further includes a bottom case, a pre-filled cartridge, and a reagent kit. The bottom case includes a reagent kit lower mounting groove, a pre-filled cartridge lower mounting groove, and pin holes. The microchannels include a first microchannel, a second microchannel, a third microchannel, a fourth microchannel, a fifth microchannel, and a sixth microchannel. The pipes include a first pipe, a second pipe, a third pipe, a fourth pipe, a fifth pipe, a sixth shunt pipe, and a seventh pipe.

[0009] Preferably, the pre-filled cartridge is pre-filled with diluent, and a sealing film is provided on the upper part, and it is installed between the pre-filled cartridge lower mounting groove and the pre-filled cartridge upper mounting groove.

[0010] Preferably, the reagent kit is pre-filled with one or more reaction reagents, and is installed between the reagent kit upper mounting groove and the reagent kit lower mounting groove. One or more reagent kit puncture tips are provided. When there are multiple ones, there is a height difference between them. The number of reagent kit puncture tips is greater than or equal to the number of reaction reagents pre-filled in the reagent kit.

[0011] Preferably, the sample loading port includes a sample chamber and a sample microfluidic port. The sample chamber is connected to the sample separation and quantification chamber through a pipe. The sample microfluidic port is located on the side of the center of the main chip.

[0012] Preferably, the sample separation and quantification chamber includes a serum chamber and a red blood cell precipitation chamber. The serum chamber is connected to the sample chamber through a first pipe, and the serum chamber is connected to the inlet of the first microchannel of the sample microchannel. The serum chamber is connected to the red blood cell precipitation chamber through a second pipe. One end of the second microchannel is connected to the middle of the second pipe, and the other end is connected to the mixing chamber. A plurality of protrusions are provided in the middle of the red blood cell precipitation chamber to prevent the film deformation that may be caused by the large span of the chamber space. The serum chamber is connected to the sample in-place detection hole and the exhaust hole respectively through a third pipe.

[0013] Preferably, the diluent loading port is a through hole penetrating both the front and back sides of the main chip, and is connected to the pre-filled cartridge on the back side of the main chip, and is connected to the diluent quantification chamber through a seventh pipe on the front side of the main chip. The diluent quantification chamber is connected to the mixing chamber through a third microchannel, and a plurality of protrusions are provided in the middle to prevent the film deformation that may be caused by the large span of the chamber space.

[0014] Preferably, the front side of the main chip further includes a quality control liquid sampling hole and a mixed liquid anti-countercurrent cavity. The quality control liquid sampling hole is connected to the dilution liquid metering cavity through a fourth pipeline, and the mixed liquid anti-countercurrent cavity is connected to the dilution liquid metering cavity, the mixing cavity, and the exhaust hole through a fifth pipeline respectively.

[0015] Preferably, the mixing cavity is connected to the sixth shunt pipeline through a fourth microchannel, and there are multiple protrusions in the middle to prevent the large span of the cavity space from possibly causing the deformation of the thin film. The sixth shunt pipeline is connected to the mixed liquid metering cavity and the mixed liquid in-place detection hole, and the mixed liquid metering cavity is provided with multiple large metering cavities and multiple small metering cavities.

[0016] Preferably, the front side of the main chip further includes a reaction liquid anti-countercurrent cavity, which is arranged between the reaction cavity and the mixed liquid metering cavity and is connected to the mixed liquid metering cavity and the reaction cavity through a sixth microchannel. The reaction cavity is connected to the colorimetric hole.

[0017] Preferably, the expansion pins correspond to the pin holes one by one and are used to fixedly connect the main chip and the bottom shell.

[0018] Preferably, the bottom of the kit lower installation groove and the prefabricated box lower installation groove is provided with an opening for lifting the kit and the prefabricated box.

[0019] Preferably, the main chip is in the shape of a sector with a central angle of 90 degrees, and a sealing film is provided on the front side. Through holes are provided at positions corresponding to the exhaust hole, the sample sampling port, and the quality control liquid sampling hole.

[0020] (III) Beneficial effects.

[0021] The utility model provides a multi-step reaction all-liquid-phase centrifugal microfluidic detection chip, which has the following beneficial effects: 1. The detection chip adopts a combined structure of a main chip, a bottom shell, and a packaging base. Spikes are arranged on the bottom shell, and a dilution liquid tank and a reaction reagent tank are provided, which can achieve multi-step all-liquid-phase reactions, making the detection of the detection chip completely consistent with the reaction system and methodology of a large biochemical analyzer. The project detection reference values are also the same, the detection results are easier to accurately evaluate and verify, the production and transportation are more convenient, and the detection is also more convenient.

[0022] 2. The quality control liquid sampling hole of the detection chip directly enters the dilution liquid metering cavity, reducing the flow path, and the chip can be subjected to standard quality control tests by the most economical method. Description of the Drawings

[0023] Figure 1 Schematic diagram of the front side of the main chip Figure 1 .

[0024] Figure 2 Schematic diagram of the front side of the main chip Figure 2 .

[0025] Figure 3 Schematic diagram of the reverse side of the main chip.

[0026] Figure 4 Schematic diagram of the bottom case.

[0027] Figure 5 Schematic diagram of the combination of the bottom case, the pre-set box and the reagent kit.

[0028] Figure 6 Schematic diagram of the chip decomposition.

[0029] Figure 7 Schematic diagram of the encapsulation base.

[0030] In the figure: 1. Main chip; 101. Sample loading port; 1011. Sample cavity; 1012. Sample microfluidic port; 102. Sample separation and quantification cavity; 1021. Serum cavity (1021); 1022. Red blood cell precipitation cavity; 103. Diluent quantification cavity; 104. Mixing cavity; 105. Reaction cavity; 106. Exhaust hole; 107. Mixing liquid in-place detection hole; 108. Sample in-place detection hole; 109. Diluent loading port; 110. Mixing liquid quantification cavity; 111. Microfluidic channel; 1111. First microfluidic channel; 1112. Second microfluidic channel; 1113. Third microfluidic channel; 1114. Fourth microfluidic channel; 1115. Fifth microfluidic channel; 112. Pipeline; 1121. First pipeline; 1122. Second pipeline; 1123. Third pipeline; 1124. Fourth pipeline; 1125. Fifth pipeline; 1126. Sixth shunt pipeline; 1127. Seventh pipeline; 113. Quality control liquid loading hole; 114. Mixing liquid reverse flow cavity; 115. Reaction liquid reverse flow cavity; 116. Reaction reagent inlet; 117. Colorimetric hole; 121. Pre-set box puncture tip; 122. Reagent kit puncture tip; 123. Expansion nail; 124. Installation groove on the pre-set box; 125. Installation groove on the reagent kit; 2. Bottom case; 21. Installation groove under the reagent kit; 22. Installation groove under the pre-set box; 23. Pin hole; 3. Pre-set box; 4. Reagent kit. Specific embodiments

[0031] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention. Embodiment

[0032] Please refer to Figures 1 - 4, the present utility model provides a centrifugal microfluidic detection chip for performing multi-step reactions in a fully liquid phase. A centrifugal microfluidic detection chip for performing multi-step reactions in a fully liquid phase includes a main chip. The main chip has front and back sides. The front side of the main chip includes a sample loading port, a sample separation and quantification chamber, a diluent quantification chamber, a mixing chamber, a reaction chamber, multiple exhaust holes, a mixed liquid in-place detection hole, a sample in-place detection hole, a diluent loading port, a mixed liquid quantification chamber, a reaction reagent inlet, a colorimetric hole, as well as multiple groups of microchannels and multiple groups of pipes for connecting and controlling fluid flow. A sealing film is provided on the front side of the main chip. It is characterized in that: the back side of the main chip includes a pre-filled cartridge puncture tip, a reagent kit puncture tip, an expansion nail, a pre-filled cartridge upper mounting groove (124), a reagent kit upper mounting groove. The above detection chip further includes a bottom case, a pre-filled cartridge, and a reagent kit. The bottom case includes a reagent kit lower mounting groove, a pre-filled cartridge lower mounting groove, and a pin hole. The microchannels include a first microchannel, a second microchannel, a third microchannel, a fourth microchannel, a fifth microchannel, and a sixth microchannel. The pipes include a first pipe, a second pipe, a third pipe, a fourth pipe, a fifth pipe, and a sixth shunt pipe.

[0033] The pre-filled cartridge is pre-filled with diluent and is provided with a sealing film on the upper part, and is installed between the pre-filled cartridge lower mounting groove and the pre-filled cartridge upper mounting groove.

[0034] The reagent kit is pre-filled with one or more reaction reagents and is installed between the reagent kit upper mounting groove and the reagent kit lower mounting groove. One or more reagent kit puncture tips are provided. When there are multiple ones, there is a height difference between them. The number of reagent kit puncture tips is greater than or equal to the number of reaction reagents pre-filled in the reagent kit.

[0035] The sample loading port includes a sample chamber and a sample microchannel port. The sample chamber is connected to the sample separation and quantification chamber through a pipe. The sample microchannel port is located on the side of the center of the main chip.

[0036] The sample separation and quantification chamber includes a serum chamber and a red blood cell precipitation chamber. The serum chamber is connected to the sample chamber through the first pipe, and the serum chamber is connected to the inlet of the first microchannel of the sample microchannel. The serum chamber is connected to the red blood cell precipitation chamber through the second pipe. One end of the second microchannel is connected to the middle of the second pipe, and the other end is connected to the mixing chamber. Multiple protrusions are provided in the middle of the red blood cell precipitation chamber to prevent the film deformation that may be caused by the large span of the chamber space. The serum chamber is connected to the sample in-place detection hole and the exhaust hole respectively through the third pipe.

[0037] The diluent loading port is a through hole penetrating both the front and back sides of the main chip, is connected to the pre-filled cartridge on the back side of the main chip, and is connected to the diluent quantification chamber through the seventh pipe on the front side of the main chip. The diluent quantification chamber is connected to the mixing chamber through the third microchannel, and multiple protrusions are provided in the middle to prevent the film deformation that may be caused by the large span of the chamber space.

[0038] The front side of the main chip further includes a quality control liquid sampling hole and a mixed liquid anti-backflow cavity. The quality control liquid sampling hole is connected to the dilution liquid metering cavity through a fourth pipeline. The mixed liquid anti-backflow cavity is connected to the dilution liquid metering cavity, the mixing cavity, and the exhaust hole respectively through a fifth pipeline.

[0039] The mixing cavity is connected to the sixth shunt pipeline through a fourth microchannel, and there are multiple protrusions in the middle to prevent the large span of the cavity space from possibly causing film deformation. The sixth shunt pipeline is connected to the mixed liquid metering cavity and the mixed liquid in-place detection hole. The mixed liquid metering cavity is provided with multiple large metering cavities and multiple small metering cavities.

[0040] The front side of the main chip further includes a reaction liquid anti-backflow cavity. The reaction liquid anti-backflow cavity is arranged between the reaction cavity and the mixed liquid metering cavity and is connected to the mixed liquid metering cavity and the reaction cavity through a sixth microchannel. The reaction cavity is connected to the colorimetric hole.

[0041] The expansion pins correspond to the pin holes one by one and are used to fixedly connect the main chip to the bottom shell.

[0042] The bottom of the reagent kit lower installation groove and the prefabricated box lower installation groove are provided with openings for lifting the reagent kit and the prefabricated box. The number of openings at the bottom of the reagent kit lower installation groove is the same as the number of reagent kits.

[0043] The main chip is in the shape of a sector with a central angle of 90 degrees. A sealing film is provided on the front side, and through holes are provided at positions corresponding to the exhaust hole, the sample sampling port, and the quality control liquid sampling hole.

[0044] The main chip and the bottom shell are fixedly connected through the cooperation of expansion pins and pin holes. The sample enters the chip by injection or capillary action through the sample sampling port. The main chip and the bottom shell are aligned through the encapsulation base, and the main chip is pressed in place. The first reagent cavity and the dilution liquid cavity in the reagent kit are lifted by the corresponding convex platforms on the encapsulation base and are punctured by the spikes on the back side of the main chip through the film of the prefabricated box and the film on the first reaction reagent kit. The complete chip body after combining the main chip and the bottom shell is loaded into the supporting instrument. The instrument drives the chip to rotate for the first time, and the reverse rotation speed is 4300 rpm for centrifugation. The sample enters the sample separation and metering cavity through centrifugal force, and the red blood cells in the sample enter the red blood cell precipitation cavity. Serum is retained in the serum cavity, and the excess serum enters the sample in-place detection hole through the third pipeline. The air in the serum is discharged through the exhaust hole. At the same time, the dilution liquid enters the dilution liquid metering cavity, and the reaction reagent enters the reaction cavity.

[0045] The instrument pauses for more than 30 seconds. Serum is detected in the sample in-place detection hole. The serum enters and fills the second microchannel through capillary action, and the dilution liquid enters and fills the third microchannel through capillary action. The instrument drives the chip to reverse-rotate again at 2500 rpm. The serum enters the mixing cavity through the centrifugal force generated by the rotation, and the dilution liquid in the dilution liquid metering cavity also enters the mixing cavity at the same time.

[0046] The instrument drives the chip to rotate forward and backward repeatedly 10 times, so that the serum and diluent in the mixing chamber are fully and evenly mixed.

[0047] The instrument pauses for more than 30 seconds, and the mixed liquid in the mixing chamber enters and fills the fourth microchannel through capillary action.

[0048] The instrument drives the chip to rotate at 200 rpm, and the mixed liquid enters the sixth shunt pipeline through centrifugal force and enters the reaction liquid metering chamber and the mixed liquid in-place detection hole, and the air in the mixed liquid is discharged through the exhaust hole.

[0049] After the mixed liquid in-place detection hole optically detects the mixed liquid, the instrument drives the chip to rotate at more than 2000 rpm, and the metered mixed liquid enters each reaction chamber respectively.

[0050] The instrument rotates forward and backward more than 10 times, the first reaction reagent and the mixed liquid are fully and evenly mixed, and the instrument pushes the encapsulation base to push the reaction reagent kit for the second time to pierce the film on the second reagent.

[0051] Let it stand for more than 3 minutes, the first reagent reacts fully with the mixed liquid, and the instrument drives the chip to rotate at more than 2000 rpm to make the second reagent enter the reaction chamber.

[0052] The instrument drives the chip to rotate forward and backward more than 10 times to fully and evenly mix the second reagent and the reactants in the first stage.

[0053] Let it stand for more than 3 minutes, and the instrument rotates slowly to continuously record the readings of the colorimetric holes communicating with each reaction chamber.

[0054] Up to 4 specimens can be detected in the same batch.

[0055] As Figures 1 - 4 As shown in Example 2, on the basis of Example 1, a quality control liquid sampling hole is also provided on the front of the main chip. On the premise of not adding samples, the quality control liquid sampling hole is directly connected to the diluent metering chamber through the fourth pipeline, and after mixing with the diluent, it enters the mixing chamber through the third microchannel, enters the mixed liquid metering chamber through the microchannel and the sixth shunt pipeline, and then enters the reaction chamber through the fifth microchannel. In this embodiment, the quality control liquid directly enters the diluent metering chamber to be mixed with the diluent, reducing the flow path, and the quality control liquid can be used to detect the deviation between the value read by the device and the standard value marked on the quality control liquid to calibrate the value read by the device, so that the device can be calibrated or used as a reference for the values read from the subsequent samples, thereby obtaining more accurate detection data at the lowest cost.

[0056] Although embodiments of the present utility model have been shown and described, those of ordinary skill in the art can understand that various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and spirit of the present utility model. The scope of the present utility model is defined by the appended claims and their equivalents.

Claims

1. A fully liquid-phase centrifugal microfluidic detection chip for performing multi-step reactions, comprising a main chip (1), the main chip (1) having front and back sides. The front side of the main chip (1) includes a sample loading port (101), a sample separation and quantification chamber (102), a diluent quantification chamber (103), a mixing chamber (104), a reaction chamber (105), multiple exhaust holes (106), a mixed liquid in-place detection hole (107), a sample in-place detection hole (108), a diluent loading port (109), a mixed liquid quantification chamber (110), a reaction reagent inlet (116), a colorimetric hole (117), as well as multiple groups of microchannels (111) and multiple groups of pipes (112) for connecting and controlling fluid flow. A sealing film is provided on the front side of the main chip (1), and it is characterized in that: The reverse side of the main chip (1) includes a pre-installed cartridge puncture tip (121), a kit puncture tip (122), an expansion nail (123), a pre-installed cartridge upper mounting groove (124), and a kit upper mounting groove (125). The above detection chip further includes a bottom shell (2), a pre-installed cartridge (3), and a kit (4). The bottom shell (2) includes a kit lower mounting groove (21), a pre-installed cartridge lower mounting groove (22), and a pin hole (23). The microfluidic channel (111) includes a first microfluidic channel (1111), a second microfluidic channel (1112), a third microfluidic channel (1113), a fourth microfluidic channel (1114), and a fifth microfluidic channel (1115). The pipeline (112) includes a first pipeline (1121), a second pipeline (1122), a third pipeline (1123), a fourth pipeline (1124), a fifth pipeline (1125), a sixth shunt pipeline (1126), and a seventh pipeline (1127).

2. The full-liquid-phase centrifugal microfluidic detection chip for performing multi-step reactions according to claim 1, wherein: The pre-installed cartridge (3) is pre-filled with a diluent and is provided with a sealing film on the upper part, and is installed between the pre-installed cartridge lower mounting groove (22) and the pre-installed cartridge upper mounting groove (125).

3. The all-liquid-phase centrifugal microfluidic detection chip for performing multi-step reactions according to claim 1, wherein: The kit (4) is pre-filled with one or more reaction reagents and is installed between the kit upper mounting groove (125) and the kit lower mounting groove (21). One or more kit puncture tips (122) are provided. When there are multiple ones, there is a height difference between them. The number of the kit puncture tips (122) is greater than or equal to the number of the reaction reagents pre-filled in the kit (4).

4. A full-liquid-phase centrifugal microfluidic detection chip for performing multi-step reactions according to claim 1 or 2, characterized in that: The sample loading port (101) includes a sample chamber (1011) and a sample microfluidic port (1012). The sample chamber (1011) is connected to the sample separation and quantification chamber (102) through a pipeline (112). The sample microfluidic port (1012) is located on the side of the center of the main chip (1).

5. A full-liquid-phase centrifugal microfluidic detection chip for performing multi-step reactions according to claim 1, characterized in that: The sample separation and quantification chamber (102) includes a serum chamber (1021) and a red blood cell precipitation chamber (1022). The serum chamber (1021) is connected to the sample chamber (1011) through the first pipeline (1121). The serum chamber (1021) is connected to the first microfluidic channel (1111) of the sample microfluidic inlet (1012). The serum chamber (1021) is connected to the red blood cell precipitation chamber (1022) through the second pipeline (1122). One end of the second microfluidic channel (1112) is connected to the middle of the second pipeline (1122), and the other end is connected to the mixing chamber (104). Multiple protrusions are provided in the middle of the red blood cell precipitation chamber (1022) to prevent the film deformation that may be caused by the large span of the cavity space. The serum chamber (1021) is respectively connected to the sample in-place detection hole (108) and the exhaust hole (106) through the third pipeline (1123).

6. The full-liquid-phase centrifugal microfluidic detection chip for performing multi-step reactions according to claim 1, characterized in that: The diluent sampling port (109) is a through-hole that penetrates both the front and back sides of the main chip (1). It is connected to the prefabricated cartridge (3) on the back side of the main chip (1), and is connected to the diluent metering chamber (103) through the seventh pipeline (1127) on the front side of the main chip (1). The diluent metering chamber (103) is connected to the mixing chamber (104) through the third microchannel (1113), and there are multiple protrusions in the middle to prevent the large span of the cavity space from possibly causing film deformation.

7. The all-liquid-phase centrifugal microfluidic detection chip for performing multi-step reactions according to claim 1, wherein: The front side of the main chip (1) further includes a quality control liquid sampling hole (113) and a mixed liquid anti-backflow chamber (114). The quality control liquid sampling hole (113) is connected to the diluent metering chamber (103) through the fourth pipeline (1124). The mixed liquid anti-backflow chamber (114) is connected to the diluent metering chamber (103), the mixing chamber (104), and the exhaust hole (106) respectively through the fifth pipeline (1125).

8. A fully liquid-phase centrifugal microfluidic detection chip for performing multi-step reactions according to claim 1, characterized in that: The mixing chamber (104) is connected to the sixth shunt pipeline (1126) through the fourth microchannel (1114), and there are multiple protrusions in the middle to prevent the large span of the cavity space from possibly causing film deformation. The sixth shunt pipeline (1126) is connected to the mixed liquid metering chamber (110) and the mixed liquid in-place detection hole (107). The mixed liquid metering chamber (110) is provided with multiple large metering chambers and multiple small metering chambers.

9. A full-liquid-phase centrifugal microfluidic detection chip for performing multi-step reactions according to claim 1, characterized in that: The front side of the main chip (1) further includes a reaction liquid anti-backflow chamber (115). The reaction liquid anti-backflow chamber (115) is arranged between the reaction chamber (105) and the mixed liquid metering chamber (110), and is connected to the mixed liquid metering chamber (110) and the reaction chamber (105) through the fifth microchannel (1115). The reaction chamber (105) is connected to the colorimetric hole (117).

10. The all-liquid-phase centrifugal microfluidic detection chip for performing multi-step reactions according to claim 1, wherein: The expansion pins (123) correspond to the pin holes (23) one by one and are used to fixedly connect the main chip (1) and the bottom case (2).

11. The full-liquid-phase centrifugal microfluidic detection chip for performing multi-step reactions according to claim 1, characterized in that: The bottom of the kit lower installation groove (21) and the prefabricated cartridge lower installation groove (22) is provided with an opening for lifting the kit (4) and the prefabricated cartridge (3).

12. A full-liquid-phase centrifugal microfluidic detection chip for performing multi-step reactions according to claim 7, characterized in that: The main chip (1) is in the shape of a sector with a central angle of 90 degrees, and a sealing film is provided on the front side. Through holes are provided at positions corresponding to the exhaust hole (106), the sample sampling port (101), and the quality control liquid sampling hole (113).

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