A chip

Abstract

The utility model relates to chip technical field, concretely relates to a kind of chip, chip is sequentially provided with first, second, third centrifugal area and detection area;Among them, first centrifugal area is provided with: sample adding pool, first pool;Second centrifugal area is provided with: first quantitative pool, first quantitative pool is connected by I type channel with sample adding pool;Third quantitative pool, third quantitative pool is connected by I type channel with first pool;Mixed cavity is provided in third centrifugal area, wherein, sample and buffer are synchronously entered mixed cavity to form mixed solution under the action of centrifugal force, and carry out reaction with target reagent in mixed cavity.The present application can improve reaction efficiency, improve the accuracy of detection result.

Description

[0001] Priority application

[0002] This application claims priority to Chinese invention patent application No. [2024109345684] "[A Chip]", filed on July 12, 2024, which is incorporated herein by reference in its entirety. Technical Field

[0003] This utility model relates to the field of microfluidic chip technology, specifically to a chip. Background Technology

[0004] POCT (Point-of-care Testing), also known as point-of-care testing or on-site rapid testing, refers to a testing method that uses portable analytical instruments and matching reagents to quickly obtain test results at the sampling site.

[0005] Immunochromatography is the most widely used point-of-care testing (POCT) technique, primarily for qualitative analysis. However, when using whole blood as the sample, it relies on whole blood quantification, which suffers from drawbacks such as inaccurate plasma quantification and inconsistent reaction efficiency between different samples, making accurate quantification difficult. Centrifugal microfluidic immunoassay can address these issues to some extent.

[0006] For example, CN111855994A discloses a POCT immunoassay chip that can perform multiple tests with a single whole blood sample addition. It includes a sample processing module for centrifuging and diluting the blood sample to obtain a plasma homogenized diluent, and a liquid-liquid reaction module for performing fluorescence immunoassay on the plasma homogenized diluent processed by the sample processing module. The liquid-liquid reaction module and the sample processing module are connected by adhesive bonding and are connected by a flow channel.

[0007] US2010 / 0151594A1 discloses an assembly of one or more microfluidic devices that together carry multiple microchannel structures, each microchannel structure including a reaction microcavity containing a solid phase having a fixed affinity ligand L, characterized in that (i) the multiple microchannel structures are divided into multiple groups, and (ii) the affinity ligand L is oriented to the same counterpart (adhesive, B) independently of the group, and (iii) these groups have a base matrix of a) the capacity of each reaction microcavity and / or the capacity / unit volume of the solid phase for adhesive B in the reaction microcavity, and / or b) equal solid phase between groups but within each group.

[0008] CN103472241A discloses a thin-film layering centrifugation device and an analytical method using the device, wherein a device for diagnosing and detecting small amounts of materials in liquids, such as laboratory-on-a-chip, protein chips, and DNA chips, is integrated into a rotatable thin-film layering body.

[0009] CN102138075A discloses a microfluidic device, which includes a microfluidic structure for providing space to contain fluid and form channels, and channels for controlling fluid flow through the microfluidic device; the microfluidic structure includes a sample chamber and a quality control chamber, wherein the sample chamber contains a sample separation unit and a testing unit for detecting samples from the supernatant using an antigen-antibody reaction, and the quality control chamber is used to identify the reliability of the test.

[0010] CN102472739A discloses a centrifugal microfluidic device for detecting analytes in liquid samples and a method for detecting analytes from liquid samples using the same device. The detection sensitivity is improved by increasing the reaction efficiency through repeated flow of the liquid sample induced by an alternating combination of capillary and centrifugal forces.

[0011] CN112763701A discloses a microfluidic detection chip and a microfluidic detection method. The chip includes at least one detection unit, which includes a sample application chamber, an immunobinding chamber, an immunodetection chamber, an immunowashing solution chamber, an immunobuffer chamber, a nucleic acid detection chamber, and a waste liquid chamber. The chip uses microfluidic technology as a carrier to combine immunological and molecular biological detection methods for tumor marker detection in a closed microfluidic chip. This enables the simultaneous detection and typing of different types of tumor markers with a small number of samples, simplifying the process and reducing the need for instruments and equipment, shortening the detection cycle, and reducing cross-contamination.

[0012] CN111273009A discloses a quantitative detection method for creatine kinase isoenzyme MB, including the preparation of creatine kinase isoenzyme MB solution, a microfluidic creatine kinase isoenzyme MB detection chip and its usage method. The microfluidic chip is characterized by a microchannel network structure, and each working unit can independently complete a single reaction or unit operation, or cooperate with other units to complete more complex work, making the entire chip a dynamic reaction system.

[0013] In other words, there are two main types of existing centrifugal microfluidic immunoassay technologies: 1) Liquid flow control is achieved through valves, but these products require high processing precision and are difficult to manufacture, and the control process of centrifugation is also relatively complex; 2) Liquid flow control is achieved through centrifugation, but these chip products are relatively limited in terms of reaction efficiency and detection accuracy. Utility Model Content

[0014] The purpose of this invention is to provide a chip that partially solves or alleviates the above-mentioned shortcomings in the prior art, thereby improving the chip's response efficiency and accuracy.

[0015] To solve the aforementioned technical problems, the present invention specifically adopts the following technical solution:

[0016] The present invention relates to a chip, characterized in that the chip is sequentially provided with a first centrifugation area, a second centrifugation area, a third centrifugation area, and a detection area;

[0017] The first centrifugation zone is equipped with:

[0018] A sample loading chamber, which is provided with a sample loading port for adding samples; a first chamber, which is used to contain buffer solution;

[0019] The second centrifuge zone is equipped with:

[0020] A first quantitative cell, which is connected to the sample dispensing cell via at least one Class I channel;

[0021] A third metering cell, wherein the third metering cell is connected to the first cell via at least one of the Class I channels;

[0022] The third centrifugation zone is equipped with:

[0023] The mixing chamber is connected to the first and third metering cells via a type II channel;

[0024] The sample in the first quantitative cell and the buffer solution in the third quantitative cell can enter the mixing chamber simultaneously under the action of centrifugal force, and mix in the mixing chamber to form a mixture;

[0025] The detection area is provided with a detection position, which is connected to the mixing chamber and is used to place the reactants. The mixture continues to enter the detection position under the action of centrifugal force.

[0026] In some embodiments, a first transition pool is provided between the sample loading pool and the first quantitative pool, the first transition pool and the sample loading pool are connected through the type I channel, and the first transition pool and the first quantitative pool are connected through the type I channel.

[0027] In some embodiments, the chip further includes: a first waste liquid pool, a first side of the first quantitative pool being connected to the first waste liquid pool via at least one of the Class I channels, the first waste liquid pool being used to contain excess liquid overflowing from the first quantitative pool, so that the first quantitative pool can quantitatively regulate the sample entering the mixing chamber.

[0028] In some embodiments, the chip further includes: a receiving cell, wherein a second side of the first metering cell is connected to the receiving cell, wherein...

[0029] The second side is provided with a first groove and a second groove that are connected to each other. The first groove is connected to the first mixing pool through the type II channel, and the depth of the first groove is greater than that of the second groove, so as to facilitate the flow and collection of the sample. The container pool is used to collect the precipitate obtained by centrifugation.

[0030] In some embodiments, the third metering pool includes: a first sub-pool and a second sub-pool connected to each other; and the bottom surface of the first sub-pool is higher than the bottom surface of the second sub-pool.

[0031] In some embodiments, the chip further includes: a second waste liquid tank, wherein the third metering tank is connected to the second waste liquid tank, wherein...

[0032] The second waste liquid pool includes a connecting pool and a storage pool, with the inlet of the connecting pool located on the first side of the first sub-pool; the second waste liquid pool is used to contain excess buffer solution overflowing from the third metering pool, so that the third metering pool can quantitatively regulate the buffer solution entering the mixing chamber.

[0033] In some embodiments, the chip further includes:

[0034] The second transition tank is connected to the second waste liquid tank through at least one of the Class II channels;

[0035] A third transition pool, wherein the third transition pool is connected to the second transition pool via at least one of the Class II channels;

[0036] The fourth quantitative cell is connected to the third transition cell through at least one of the type II channels; the second mixing cell is connected to the fourth quantitative cell through the type II channels; the fourth quantitative cell is also connected to the detection position through the type I channels.

[0037] In some embodiments, the second waste liquid tank, the second transition tank, the third transition tank, and the fourth quantitative tank are arranged in a direction that moves away from the centrifuge center in sequence.

[0038] In some embodiments, the type II channel is a U-shaped capillary channel, and the U-shaped opening of the U-shaped capillary channel is oriented away from the centrifugal center.

[0039] In some embodiments, the fourth metering chamber includes a first communicating cavity, a second communicating cavity, and a metering cavity; wherein,

[0040] The first connecting cavity is connected to the third transition pool and the second mixing pool respectively; the inlet end of the metering cavity is connected to the outlet end of the first connecting cavity, and the inlet end of the second connecting cavity is connected to the inlet end of the metering cavity.

[0041] In some embodiments, the chip is provided with at least one air hole connected to the mixing chamber to maintain air pressure balance and ensure smooth liquid flow.

[0042] In some embodiments, the chip is provided with a control area for setting a control substance.

[0043] Beneficial technical effects:

[0044] This invention designs an automated mixing circuit based on centrifugation. This circuit enables automated, synchronous quantitative mixing. Specifically, the sample and buffer solution are centrifuged and then fed into the first and third quantitative cells respectively, achieving quantification of the sample and buffer solution. The first and third quantitative cells are connected to the first side of the first mixing cell via two U-shaped capillary channels, allowing the sample and buffer solution to enter the first mixing cell synchronously under centrifugal force. Multiple drainage zones in the first mixing cell thoroughly mix the sample and buffer solution to dilute the sample before it enters the second mixing cell. The impact of multiple drainage zones in the second mixing cell ensures a thorough reaction between the sample and antibody. This mixing structure automates the quantification of buffer solution and sample through centrifugation, eliminating the need for manual control and reducing the workload of testing personnel. Synchronous, automated quantitative mixing at specific times before antibody reaction allows for a more complete reaction process, improving reaction efficiency and detection sensitivity.

[0045] On the other hand, this invention also designs an automatic splitting branch based on multi-stage centrifugation. Specifically, the buffer solution pre-stored in the first pool automatically enters the third quantitative pool for quantification under centrifugal force. Simultaneously, excess buffer solution enters the second waste pool, and through multiple centrifugations, sequentially enters the second transition pool, the third transition pool, and the fourth quantitative pool, finally entering the detection station for cleaning under centrifugal force. In other words, automatic splitting means that the buffer solution can be centrifuged to enter the third quantitative pool and the second waste pool separately. The buffer solution in the third quantitative pool enters the mixing branch, and the buffer solution in the second waste pool enters the automatic cleaning process. The automatic splitting branch can further reduce the workload of testing personnel. A single addition of buffer solution can be used for both sample dilution and mixing, as well as for automatic cleaning. Furthermore, the design of the first pool can also be used for pre-storing a liquid sac (specifically, a liquid sac is provided in the first pool to hold the buffer solution; the liquid sac has an opening, which is sealed by a sealing strip), further reducing the workload of testing personnel.

[0046] This invention combines automated synchronous quantitative mixing and automatic cleaning through a dual-branch design, further improving the sensitivity and specificity of detection. Attached Figure Description

[0047] To more clearly illustrate the technical solutions in the embodiments of this utility model or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. In all the drawings, similar elements or parts are generally identified by similar reference numerals. The elements or parts in the drawings are not necessarily drawn to scale. Obviously, the drawings described below are some embodiments of this utility model. For those skilled in the art, other drawings can be obtained based on these drawings without any creative effort.

[0048] Figure 1a This is a first schematic diagram of the chip structure in an exemplary embodiment of the present invention;

[0049] Figure 1b This is a second schematic diagram of the chip structure in an exemplary embodiment of the present invention;

[0050] Figure 2 This is a third schematic diagram of the chip structure in an exemplary embodiment of the present invention;

[0051] Figure 3 This is a schematic diagram of the first partial structure of the chip in an exemplary embodiment of the present invention;

[0052] Figure 4a This is a schematic diagram of the second partial structure of the chip in an exemplary embodiment of the present invention;

[0053] Figure 4b This is a schematic diagram of the third partial structure of the chip in an exemplary embodiment of the present invention;

[0054] Figure 4c This is a schematic diagram of the fourth partial structure of the chip in an exemplary embodiment of the present invention;

[0055] Figure 4d This is a schematic diagram of the fifth partial structure of the chip in an exemplary embodiment of the present invention;

[0056] Figure 5 This is a schematic diagram of the sealing structure of a chip in an exemplary embodiment of the present invention;

[0057] Figure 6 This is a schematic diagram of the chip structure in an exemplary embodiment of the present invention.

[0058] Figure 7 This is a functional schematic diagram of the chip in an exemplary embodiment of the present invention;

[0059] Figure 8This is a schematic diagram of the chip's workflow in an exemplary embodiment of the present invention;

[0060] Figure 9 This is a schematic diagram of the liquid flow state inside the chip in an exemplary embodiment of the present invention;

[0061] Figure 10 This is a schematic diagram of the packaging strip and sample feeding hole in an exemplary embodiment of the present invention;

[0062] Figure 11 This is a schematic diagram of the channel layer structure of the centrifugal microfluidic immunoassay reagent tray in another exemplary embodiment of the present invention;

[0063] Figure 12 This is a schematic diagram of the liquid bladder structure in another exemplary embodiment of the present invention;

[0064] Figure 13 This is a schematic diagram of the test strip structure in another exemplary embodiment of the present invention;

[0065] Figure 14 This is a schematic diagram of the control test strip structure in another exemplary embodiment of the present invention;

[0066] Figure 15 This is a schematic diagram of the overall structure of the centrifugal microfluidic immunoassay reagent tray in another exemplary embodiment of the present invention;

[0067] Figure 16 This is a repeatable result in yet another exemplary embodiment of the present invention;

[0068] Figure 17 The methodological comparison results are for another exemplary embodiment of this utility model.

[0069] Summary of attached image labels:

[0070] A set of reference numerals in the attached diagram are as follows: 1 is the first centrifugation zone; 10 is the sample loading chamber; 101 is the sample loading port; 12 is the first channel; 13 is the second channel; 14 is the first quantitative chamber; 141 is the first groove; 142 is the second groove; 15 is the first waste liquid chamber; 16 is the receiving chamber; 17 is the first transition chamber; 20 is the first chamber; 20A is the sealing strip; 21 is the third quantitative chamber; 211 is the first sub-chamber; 212 is the second sub-chamber; 22 is the second waste liquid chamber; 221 is the storage chamber; 222 is the connecting chamber; 30 is the mixing chamber. 31 is the fourth channel, 32 is the fifth channel, 33 is the first mixing tank, 331 is the first drainage zone, 332 is the second drainage zone, 333 is the third drainage zone, 34 is the second mixing tank, 344 is the fourth drainage zone, 345 is the fifth drainage zone, 346 is the sixth drainage zone, and 35 is an air vent; 41 is the second transition tank, 43 is the fourth quantitative tank, 431 is the second connecting cavity, 432 is the quantitative cavity, 433 is the first connecting cavity, 45 is the third transition tank, 46 is the third waste liquid tank; and 51 is the detection position.

[0071] Another set of attached diagrams is labeled as follows: 1 is the cover plate layer, 101 is the sample dispensing port, 102 is the positioning hole, and 103 is the buffer dispensing hole; 2 is the channel layer, 201 is the sample dispensing chamber, 202 is the plasma quantification chamber, 203 is the erythrocyte storage chamber, 204 is the capillary channel, 205 is the fluid sac placement chamber (buffer dispensing chamber), 206 is the buffer quantification chamber, 207 is the buffer transition chamber, 208 is the positioning hole, 209 is the vent, 210 is the reaction chamber, 211 is the chromatography quantification chamber, and 212 is the residual liquid storage chamber. The test strip is placed in the storage chamber (waste liquid chamber). 213 is the test strip placement chamber, and 214 is the control test strip placement chamber. 3 is the liquid sac, 301 is the sealing strip, and 302 is the liquid sac shell. 4 is the test strip, 401 is the base plate, 402 is the reaction pad, and 403 is the absorbent pad. 5 is the control test strip, 501 is the base plate, and 502 is the biofilm. 601 is the fluorescent microsphere, 602 is antibody 1, 603 is the analyte-antigen in plasma, 604 is antibody 2, 605 is biotin, and 606 is streptavidin. Detailed Implementation

[0072] To make the objectives, technical solutions, and advantages of the embodiments of this utility model clearer, the technical solutions of the embodiments of this utility model will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this utility model. All other embodiments obtained by those skilled in the art based on the embodiments of this utility model without creative effort are within the scope of protection of this utility model.

[0073] In this document, suffixes such as "module," "part," or "unit" used to denote elements are used only for the purpose of illustrative purposes and have no specific meaning in themselves. Therefore, "module," "part," or "unit" can be used interchangeably.

[0074] In this document, the terms "upper," "lower," "inner," "outer," "front," "rear," "one end," and "the other end," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are used only for the convenience of describing this utility model and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this utility model. Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0075] In this document, unless otherwise explicitly specified and limited, the terms "installed," "equipped with," and "connected," etc., should be interpreted broadly. For example, "connected" can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection, a direct connection, or an indirect connection through an intermediate medium; it can be a connection within two components. Those skilled in the art can understand the specific meaning of the above terms in this utility model based on the specific circumstances.

[0076] In this document, "and / or" includes any and all combinations of one or more of the listed related items.

[0077] In this article, "multiple" means two or more, that is, it includes two, three, four, five, etc.

[0078] It should be noted that, in this document, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes that element.

[0079] As used in this specification, the term "about" typically means + / - 5% of the value, more typically + / - 4%, more typically + / - 3%, more typically + / - 2%, even more typically + / - 1%, even more typically + / - 0.5% of the value.

[0080] In this specification, certain embodiments may be disclosed in a range-bound format. It should be understood that this "range-bound" description is merely for convenience and brevity and should not be construed as a rigid limitation on the disclosed range. Therefore, the description of a range should be considered as having specifically disclosed all possible subranges and the individual numerical values ​​within those ranges. For example, a description of the range 1-6 should be considered as having specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., and the individual numbers within those ranges, such as 1, 2, 3, 4, 5, and 6. This rule applies regardless of the breadth of the range.

[0081] Glossary

[0082] In this article, unless otherwise explicitly stated and defined, the two ends of the channel (the first end and the second end) are also referred to as the inlet and outlet, respectively, along the direction of liquid (such as sample, buffer solution, etc.) flow in the channel.

[0083] In this document, a pool (or cavity) refers to a spatial region capable of containing liquid, and the first side of a pool (or cavity) (in some embodiments, it may also refer to the first end of the pool or cavity) refers to the side of the chip closer to the centrifugation center during the centrifugation process. In some embodiments described herein, pools or cavities may be used interchangeably. It should be understood that the description of pools or cavities is merely for illustrative purposes and should not be construed as indicating or implying differences in spatial regions.

[0084] In this article, unless otherwise explicitly stated and defined, width refers to the length in a direction perpendicular or approximately perpendicular to the radial direction (or, in other words, the centrifugal radius).

[0085] In this article, unless otherwise explicitly stated or limited, length refers to length in the radial direction.

[0086] In this article, depth refers to the length along a vertical direction that is perpendicular or approximately perpendicular to the surface of the chip.

[0087] Example 1

[0088] This utility model provides a chip, which includes a first centrifugation area, a second centrifugation area, a third centrifugation area, and a detection area (e.g., ...). Figures 1a-1b As shown), where,

[0089] The first centrifuge zone is equipped with:

[0090] The sample loading chamber 10 is provided with a sample loading port 101 for adding samples; the first chamber 20 is used to contain buffer solution.

[0091] The sample in the sample loading chamber 10 and the buffer solution in the first chamber can enter the second centrifugation zone under the action of centrifugal force.

[0092] The second centrifuge zone is equipped with:

[0093] A first quantitative cell 14 is connected to the sample loading cell 10 via at least one Class I channel and is used to receive samples from the sample loading cell 10.

[0094] The third quantitative pool 21 is connected to the first pool 20 through at least one Class I channel and is used to receive buffer solution from the first pool 20.

[0095] The third centrifugation zone is equipped with:

[0096] A mixing chamber 30 includes a first mixing pool 33 and a second mixing pool 34. A connecting portion is provided between the second end of the first mixing pool 33 and the first end of the second mixing pool 34. The width of the connecting portion (the width of the connecting portion refers to its size in a direction perpendicular or approximately perpendicular to the flow direction of the liquid) is smaller than the width of the second end of the first mixing pool 33 and / or the width of the first end of the second mixing pool 34. Furthermore, the first mixing pool 33 is connected to the first metering pool 14 and the third metering pool 21 via Class II channels.

[0097] For example, in some embodiments, the sample in the first quantitative cell 14 and the buffer solution in the third quantitative cell 21 can at least partially fill the two type II channels under capillary action when at rest. However, due to the confinement effect of the two type II channels, the liquid inside will not continue to enter the first mixing cell 33. Only under the action of centrifugal force can the liquid in the two type II channels enter the first mixing cell 33 simultaneously to mix and form the first mixture.

[0098] Furthermore, the chip is provided with at least one air hole 35 connected to the mixing chamber 30 to maintain the air pressure balance inside the chip and ensure smooth liquid flow.

[0099] In some embodiments, the direction of liquid flow in the type II channel may be: first segment 01 → second segment 02 → third segment 03. Figure 4aSpecifically, the inlet end of the Type II channel extends towards the centrifuge center to form a first segment 01, and the outlet end of the Type II channel extends towards the centrifuge center to form a third segment 03. The first segment 01 and the third segment 03 are connected by a second segment 02. The first segment 01 is connected to the first groove of the first metering tank 14, and the liquid in the first segment 01 gradually flows towards the centrifuge center. The second segment 02 connects the first segment 01 and the third segment 03, acting as a deflector to facilitate the smooth flow of liquid from the first segment 01 into the third segment 03. The third segment 03 is connected to the first mixing tank 33, and the liquid in the third segment 03 gradually flows away from the centrifuge center. Furthermore, the distance from the inlet end of the Type II channel to the centrifuge center is less than the distance from the outlet end to the centrifuge center.

[0100] Furthermore, the first mixing tank 33 contains a first drainage zone 331, a second drainage zone 332, and a third drainage zone 333 (e.g., ...). Figure 4b (As shown). The first diversion zone 331 is connected to the first and third metering pools, and the liquid entering this zone can undergo preliminary mixing; the second diversion zone 332 connects the first and third diversion zones and can be used to extend the liquid flow path and enhance mixing efficiency; the liquid can be guided by the second diversion zone 332 into the third diversion zone 333 and further mixed under the impact.

[0101] Specifically, the inlet end of the first drainage zone 331 is connected to the first metering pool 14 and the third metering pool 21 respectively; the inlet end and outlet end of the second drainage zone 332 are connected to the outlet end of the first drainage zone 331 and the inlet end of the third drainage zone 333 respectively, thereby guiding the liquid in the first drainage zone 331 to enter the third drainage zone 333 after at least one turning.

[0102] Preferably, the width of the inlet end of the second drainage area 332 (e.g.) Figure 3 The width of the outlet end of the first drainage area 331 is less than the width of the outlet end of the second drainage area 332, which is less than the width of the inlet end of the third drainage area 333.

[0103] Preferably, the inlet and outlet of the third drainage zone 333 are arranged in a direction that moves away from the centrifugal center, and the outlet is arranged in a direction that moves away from the inlet (e.g., the inlet and outlet are respectively arranged at both ends of the third drainage zone).

[0104] In this embodiment, the liquid is guided to change direction at least once under the impact of the chamber wall by the diversion setting of multiple drainage areas, so as to enable the sample and buffer to be mixed more thoroughly when passing through the first mixing pool briefly, so that the first mixture can react more efficiently with the antibody in the second mixing pool 34.

[0105] The first mixture enters the second mixing tank through the connecting part under the action of centrifugal force. The first mixture reacts with the target reagent in the second mixing tank to form the second mixture.

[0106] Furthermore, a fourth drainage zone 344, a fifth drainage zone 345, and a sixth drainage zone 346 (e.g., ...) exist in the second mixing tank 34. Figure 4b (As shown). The fourth drainage zone 344 is connected to the third drainage zone 333 through a connecting part. The length of its inlet end is relatively smaller than the width of its outlet end, so as to promote the re-mixing of liquids under the squeezing action to a certain extent. The liquid in the fourth drainage zone 344 is rushed into the fifth drainage zone 345 under the centrifugal action and further mixed under the impact action. The sixth drainage zone 346 can promote the further mixing of some liquids.

[0107] Specifically, the inlet end of the fourth drainage zone 344 is connected to the outlet end of the third drainage zone 333 via a connecting portion; the outlet end of the fourth drainage zone 344 is connected to the inlet end of the fifth drainage zone 345, and the width of the outlet end of the fourth drainage zone 344 is smaller than the width of the inlet end of the fifth drainage zone 345; and the fifth drainage zone 345 is also connected to the fourth metering tank 43 via the type II channel; wherein, the inlet end of the fourth drainage zone 344 extends in a direction gradually moving away from the connecting portion (i.e., along...). Figure 3 (Extending in the direction L) forms a channel with a certain width, and the size of the channel (i.e., in relation to the width direction L) (see...) Figure 3 The length in the direction perpendicular or approximately perpendicular to the liquid inlet of the fifth drainage area 345 is less than the width of the liquid inlet end of the fifth drainage area 345; preferably, antibodies are provided in the direction away from the liquid inlet end of the fifth drainage area 345.

[0108] Further, see Figure 4cAs shown, a sixth drainage zone 346 connected to the fifth drainage zone 345 is also provided. The sixth drainage zone 346 is located on the side away from the liquid outlet end of the fifth drainage zone 345, so that the liquid that is deflected by the impact of the wall of the fifth drainage zone 345 can continue to enter the sixth drainage zone 346 and undergo delayed mixing under the impact of the sixth drainage zone 346. Specifically, the sixth drainage zone 346 can be a region extending from the side away from the liquid outlet end of the fifth drainage zone 345 in a direction close to the centrifugal center (or, a region protruding towards the centrifugal center).

[0109] Through the spatial synergy of the fourth, fifth and sixth mixing zones, the liquid in the fourth drainage zone 344 can be guided to enter the sixth drainage zone 346 after at least three turns, and fully react with the antibody under the impact generated by the cooperation of multiple drainage zones;

[0110] Preferably, the inlet and outlet of the fifth drainage zone 345 are arranged in a direction that moves away from the centrifugal center, and the outlet is arranged in a direction that moves away from the inlet (e.g., the inlet and outlet are respectively located at the two ends of the fifth drainage zone 345).

[0111] In this embodiment, the liquid is guided to undergo at least three turns under the impact of the chamber wall through the diversion settings of multiple drainage zones. This ensures that the first mixture formed can be fully mixed with the antibody during its brief passage through the second mixing chamber, allowing the second mixture to react more efficiently with the biomaterial with capture function at the detection site. In other words, the mixing and reaction process in this embodiment can be achieved through only one centrifugation process, which is simpler and results in higher detection efficiency.

[0112] Furthermore, the first side (at least a portion) of the second mixing pool 34 is disposed opposite to the outer edge of the receiving pool 16 (for example, the length of at least a portion of the side of the second mixing pool on the first side gradually decreases along the direction W from its first end to its second end); their combined effect can improve the utilization rate of planar space to a certain extent.

[0113] Furthermore, at least a portion of the liquid in the mixing chamber 30 will undergo at least one (e.g., 1, 2, 3, 4...) diversion (e.g., ... Figure 4c As shown), and enhances mixing efficiency under the corresponding steering (impact) action.

[0114] In some embodiments, the drainage area may also be referred to as the mixing area.

[0115] The detection area is equipped with:

[0116] The detection station is used to set the reactants, and the second mixture eventually enters the detection station under the action of centrifugal force.

[0117] In some embodiments, the chip further includes a first transition cell 17 disposed between the sample loading cell 10 and the first quantitative cell 14, the first transition cell 17 being connected to the sample loading cell 10 via at least one Class I channel, and the first transition cell 17 being connected to the first quantitative cell 14 via a Class I channel.

[0118] In some embodiments, the chip further includes a first waste liquid pool 15, which is connected to a first side of the first metering pool 14 via at least one Class I channel.

[0119] The first waste liquid pool 15 is used to contain excess sample overflowing from the first quantitative pool 14, so that the first quantitative pool 14 can quantitatively regulate the sample entering the mixing chamber 30.

[0120] In some embodiments, the chip further includes a containment pool 16 for collecting precipitates obtained by centrifugation and is connected to a second side of the first quantitative pool 14.

[0121] Furthermore, the second side of the first quantitative cell 14 is provided with a first groove 141 and a second groove 142 that are connected to each other. The first groove 141 is connected to the first mixing cell through a type II channel. And the depth of the first groove is greater than that of the second groove 142, so as to facilitate the drainage and collection of the sample.

[0122] In some embodiments, the third metering pool 21 includes a first sub-pool 211 and a second sub-pool 212 that are connected to each other, and the bottom surface of the first sub-pool is higher than the bottom surface of the second sub-pool.

[0123] In some embodiments, the chip further includes a second waste liquid pool 22, which is connected to the third quantitative pool 21.

[0124] Furthermore, the second waste liquid tank 22 includes a connecting tank 222 and a storage tank 221, with the inlet of the connecting tank 222 located on the first side of the first sub-tank 211. The second waste liquid tank 22 is used to contain excess buffer solution overflowing from the third metering tank 21, so that the third metering tank 21 can quantitatively regulate the buffer solution entering the mixing chamber 30.

[0125] The chip described in this embodiment also includes a second transition cell 41, a third transition cell 45, and a fourth quantitative cell 43.

[0126] The second transition pool 41 is connected to the second waste liquid pool 22 through at least one Class II channel; the third transition pool 45 is connected to the second transition pool 41 through at least one Class II channel; the fourth quantitative pool 43 is connected to the third transition pool 45 through at least one Class II channel, connected to the second mixing pool 34 through at least one Class II channel, connected to the third waste liquid pool 46 through at least one Class I channel, and also connected to the detection position through a Class I channel.

[0127] Furthermore, the second waste liquid pool 22, the second transition pool 41, the third transition pool 45, and the fourth quantitative pool 43 are arranged on the chip in a direction that moves away from the centrifugation center in sequence.

[0128] Preferably, the second waste liquid tank 22, the second transition tank 41, and the third transition tank 45 are arranged on the same straight line.

[0129] Preferably, the above-mentioned Type II channels are all U-shaped capillary channels, and the U-shaped opening of the U-shaped capillary channel is set in a direction away from the centrifugal center so as to automatically enter the next chamber under the action of centrifugal force.

[0130] In some embodiments, the fourth metering chamber 43 includes a first connecting cavity 433, a second connecting cavity 431, and a metering cavity 432. The first connecting cavity 433 is connected to the third transition chamber 45 and the second mixing chamber 34, respectively; the inlet end of the metering cavity 432 is connected to the outlet end of the first connecting cavity 433, and the inlet end of the second connecting cavity 431 is connected to the inlet end of the metering cavity 432.

[0131] When the second mixture moves to the fourth metering tank 43, the binding reaction has ended and the capture reaction has not started; the irregular structure of the first connecting cavity 433 ensures that the second mixture enters the metering cavity smoothly and does not fall directly into the third waste liquid tank 46.

[0132] Preferably, the first connecting cavity 433 is configured to have a narrow channel, and the narrow channel is connected to the metering cavity 432 through a first connecting part, so as to guide the reaction liquid in the narrow channel directly into the metering cavity 432; the first end of the metering cavity 432 is connected to the second connecting cavity 431 through a second connecting part, thereby guiding the reaction liquid overflowing from the metering cavity 432 into the second connecting cavity 431;

[0133] Preferably, the first connecting portion and the second connecting portion are arranged at intervals so that the first and second connecting portions have a certain distance between them;

[0134] The independent arrangement of the two connecting parts divides the fourth quantitative cell into three relatively independent chambers (equivalent to forming an irregular structure). The different connecting parts guide the reaction liquid to enter the quantitative chamber first, while ensuring that excess liquid can be smoothly discharged to meet the quantitative detection requirements.

[0135] A height difference is provided between the metering cavity 432 and the first connecting cavity 433, with the bottom surface of the metering cavity 432 being higher than that of the first connecting cavity 433, in order to facilitate the metering of the second mixture.

[0136] After the capture reaction in the detection zone is completed, centrifugation is performed again, allowing the remaining buffer solution in the third transition cell 45 to flow further into the quantitative chamber of the fourth quantitative cell 43. This allows the remaining buffer solution to enter the detection zone under centrifugal force, washing away the uncaptured second mixture, reducing the background signal, and thus improving the sensitivity and accuracy of the reaction.

[0137] In this invention, the chip also has a control area independent of the detection area, used for setting a control substance. The control area does not participate in the reaction, thus ensuring good repeatability.

[0138] On one hand, this embodiment designs an automated mixing branch based on centrifugation. This branch enables automated, synchronous quantitative mixing. Specifically, the sample and buffer solution are centrifuged and enter the first quantitative chamber 14 and the third quantitative chamber 21, respectively, achieving quantification of the sample and buffer solution. The first quantitative chamber 14 and the third quantitative chamber 21 are connected to the first side of the first mixing chamber 33 via two U-shaped capillary channels, allowing the sample and buffer solution to enter the first mixing chamber 33 synchronously under centrifugal force. Under the action of multiple drainage zones in the first mixing chamber 33, the sample and buffer solution are thoroughly mixed to dilute the sample, which then enters the second mixing chamber 34. Under the impact of multiple drainage zones in the second mixing chamber 34, the sample and antibody react fully. This mixing structure automates the quantification of buffer solution and sample through centrifugation, eliminating the need for manual control and reducing the workload of testing personnel. Synchronous, automated quantitative mixing at specific times before reaction with the antibody allows for a more complete reaction process, improving reaction efficiency and detection sensitivity.

[0139] On the other hand, this embodiment designs an automatic splitting branch based on multi-stage centrifugation. Specifically, the buffer solution pre-stored in the first pool automatically enters the third quantitative pool 21 for quantification under centrifugal force. Simultaneously, excess buffer solution enters the second waste pool 22, and through multiple centrifugations, sequentially enters the second transition pool 41, the third transition pool 45, and the fourth quantitative pool 43, finally entering the detection station for cleaning under centrifugal force. In other words, automatic splitting means that the buffer solution can be centrifuged to enter the third quantitative pool 21 and the second waste pool 22 respectively. The buffer solution in the third quantitative pool 21 enters the mixing branch, and the buffer solution in the second waste pool 22 enters the automatic cleaning process. The automatic splitting branch can further reduce the workload of testing personnel. A single addition of buffer solution can be used for both sample dilution and mixing, as well as for automatic cleaning. Furthermore, the design of the first pool can also be used for pre-storing a liquid sac (specifically, a liquid sac is provided in the first pool to hold the buffer solution; the liquid sac has an opening, which is sealed by a sealing strip), further reducing the workload of testing personnel.

[0140] This embodiment combines automated synchronous quantitative mixing and automatic cleaning through a dual-branch design, further improving the sensitivity and specificity of detection.

[0141] Beneficial technical effects:

[0142] A separate C-line is set up so that the control line results are stable and not affected by the reaction efficiency.

[0143] Multiple tests can be performed using the same C-line, making it more economical and efficient;

[0144] The mixing chamber allows for thorough mixing, resulting in high reaction efficiency and good sensitivity.

[0145] Using avidin bands for capture simplifies reagent processing. Different assays only require adding different reagents to the detection chamber, simplifying the process and making it suitable as a universal immunoassay platform. It also features an automated washing step, resulting in better reaction specificity.

[0146] Furthermore, the diluent and cleaning solution in this invention share a single liquid bladder, making it more practical.

[0147] See Figure 1a and Figure 1bAs shown, this utility model provides a chip, which is sequentially arranged with a first centrifugation zone, a second centrifugation zone, a third centrifugation zone, a fourth centrifugation zone, and a detection zone. Under multi-stage centrifugation, the liquid sequentially passes through the first centrifugation zone, then the second, third, and fourth centrifugation zones, gradually mixing the sample and buffer solution. Finally, the resulting mixture enters the detection zone to react with the reactants, and the reaction result between the mixture and the reactants is used as the test result.

[0148] The first centrifugation zone I (also known as the sample loading zone) is equipped with:

[0149] The sample loading chamber 10 is provided with a sample loading port 101 for sample loading (e.g., the sample may be whole blood); the first chamber 20 is used to contain buffer solution;

[0150] The second centrifugation zone (also known as the quantification zone) contains:

[0151] The first quantitative cell 14 is connected to the sample addition cell 10 through at least one channel;

[0152] A container 16 is connected to the first quantitative container 14; wherein, the container 16 can store red blood cells obtained after whole blood separation;

[0153] The third metering pool 21 is connected to the first pool 10 through at least one channel;

[0154] The second waste liquid tank 22 is connected to the third metering tank 21;

[0155] See Figure 2 As shown, the third centrifuge zone III is equipped with:

[0156] The mixing chamber 30 is connected to the first metering cell 14 through the fourth channel 31 and to the third metering cell 21 through the fifth channel 32;

[0157] The fourth centrifuge zone IV is equipped with:

[0158] The second transition pool 41 is connected to the second waste liquid pool 22 through the seventh channel;

[0159] The fourth metering pool 43 is connected to the mixing chamber 30 through the sixth channel, and is also connected to the second transition pool 41 through at least one channel.

[0160] The detection zone V is equipped with:

[0161] The detection position 51 is used to set a reactant (preferably a test strip), wherein a chemical reaction can occur when the mixture to be tested comes into contact with the reactant, and is then used to characterize the physical or chemical properties of the mixture or sample through chemical reaction phenomena (such as color change of the test strip).

[0162] Furthermore, in some embodiments, the chip further includes:

[0163] The third channel has its first end (i.e., the inlet) connected to the first metering tank 14, and its second end (i.e., the outlet) connected to the first waste liquid tank 15.

[0164] The first waste liquid pool 15 is used to contain excess liquid overflowing from the first quantitative pool 14, so that the first quantitative pool 14 can quantitatively control the sample entering the subsequent reaction space.

[0165] The first end of the third channel is located on the first side of the first quantitative pool 14.

[0166] Referring to Figure 1, in some embodiments, it further includes:

[0167] A first transition pool 17 is also provided between the sample loading pool 10 and the first quantitative pool 14. The first transition pool 17 is connected to the sample loading pool 10 through at least one first channel 12, and the first transition pool 17 is connected to the first quantitative pool 14 through at least one second channel 13.

[0168] In this embodiment, the setting of the first transition pool can prevent the fourth channel 31 from being opened in advance, that is, to prevent the liquid in the first quantitative pool 14 from being completely drained after centrifugation is started, which would lead to quantitative failure.

[0169] See Figure 3 As shown, in some embodiments, the third metering pool 21 includes: a first sub-pool 211 and a second sub-pool 212 that are connected to each other; and the depth H1 of the first sub-pool 211 is less than the depth H2 of the second sub-pool 212.

[0170] Preferably, H1 is 0.2mm-6mm, and 212 depth H2 is 0.3mm-9mm. In general, however, H2 is 0.3mm-9mm.

[0171] See Figure 3 As shown, in some embodiments, the second waste liquid pool 22 includes a connecting pool 222 and a storage pool 221. The inlet of the connecting pool 222 is located on the first side of the third quantitative pool 21. The connecting pool 222 is used to connect the storage pool 221 and the third quantitative pool 21 in space, and can also limit excessive liquid (such as buffer solution) from flowing into the storage pool 221, thereby affecting the quantitative effect of the third quantitative pool.

[0172] The movement path of the liquid from the third metering tank 21 into the storage tank 221 is as follows: Figure 3 The direction indicated by the middle arrow.

[0173] In some embodiments, the connecting pool 222 can be configured as a pipe structure.

[0174] Preferably, one or more channels in this invention are capillary channels.

[0175] See Figure 2 As shown, in some embodiments, the mixing chamber includes:

[0176] The first mixing tank 33 is connected to the first quantitative tank 14 and the third quantitative tank 21 through the fourth channel 31 and the fifth channel 32, respectively.

[0177] The second mixing tank 34 is connected to the first mixing tank 33 via at least one channel.

[0178] As shown in the figure, the two mixing chambers in this invention can adopt an irregular shape design, which forces the sample and buffer solution from the first quantitative chamber 14 and the third quantitative chamber 21 to come into contact before reaching the second mixing chamber 34. The structure protruding to the lower left in the center of the first mixing chamber 33 makes the flow path of the liquid in the structure longer, thus increasing the mixing effect.

[0179] In this embodiment, the buffer solution and the sample can be initially mixed in the first mixing chamber 33, and then further mixed in the second mixing chamber 34.

[0180] Preferably, the second mixing chamber contains pre-stored antibodies for mixing with the mixture. For example, when the sample is blood, the antibodies include biotin conjugates and microsphere conjugates. All antibodies are pre-dried.

[0181] See Figure 3 As shown, in some embodiments, the second side of the first metering cell 14 is provided with:

[0182] The first groove 141 and the second groove 142, wherein the depth of the first groove 141 is greater than that of the second groove 142, are used to guide and collect the liquid.

[0183] In some embodiments, the inner diameter of the second side of the first metering pool 14 (i.e. the side communicating with the receiving pool 16) gradually decreases (e.g., it is configured in a tapering shape).

[0184] In some embodiments, when the chip is placed in a centrifuge, the distance between the first end of the fourth channel 31 and the centrifugation center (or rotation center) is less than the distance between the second end of the fourth channel 31 and the centrifugation center.

[0185] In other words, during centrifugation, the centrifugal force at the first end of the fourth channel is less than the centrifugal force at its second end.

[0186] In some embodiments, see Figure 1- Figure 4d As shown, at least one channel (such as the fourth channel 31, the fifth channel 32, the sixth channel, the seventh channel, etc.) can respectively play the roles of flow restriction and connection in different centrifugation stages. Correspondingly, these channels can also be called flow restriction channels.

[0187] For example, see Figure 4a The fourth channel 31 is a capillary channel, and is sequentially arranged with a first section 01, a second section 02, and a third section 03 from its first end to its second end. The first section 01 is connected to the first metering tank 14 and extends gradually towards the centrifugal center. The second section 02 guides the rotation of the liquid and connects to the third section 03, which is connected to the first mixing tank 31. When the centrifugal force is small, the liquid in the first metering tank 14 will partially enter the first section 01 under the action of capillary force, but will be restricted by the second section 02 and unable to continue moving into the mixing chamber.

[0188] In some embodiments, the second mixing tank 34 is provided with air holes.

[0189] In some embodiments, the chip further includes a third transition cell 45, which is connected to a fourth quantitative cell 43 via at least one channel (also referred to as a current limiting channel).

[0190] In some embodiments, the reactants may be in a liquid or solid state. For example, the reactants may be provided via test strips, such as test strips pre-coated with streptavidin.

[0191] In some embodiments, the detection position 51 is also provided with a limiting element (such as a protrusion provided along the test strip).

[0192] In some embodiments, the chip further includes a third waste liquid tank 46, which is connected to a fourth metering tank 43.

[0193] In some embodiments, the fourth metering cell 43 includes:

[0194] The first communicating cavity 433 is connected to the third transition pool 45 through at least one channel;

[0195] A metering cavity 432 that is connected to the first communicating cavity 433;

[0196] The second connecting cavity 431 is connected to the first connecting cavity 433 and the metering cavity 432 respectively.

[0197] Preferably, the liquid inlet of the second connecting cavity 431 is located at the connection between the first connecting cavity 433 and the metering cavity 432.

[0198] In some embodiments, a control area 52 (i.e., an independent C-line) is provided.

[0199] The chip provided in this embodiment is preferably suitable for the double antibody sandwich method. It contains two types of antibodies in its internal mixing chamber: one monoclonal antibody conjugated with biotin and the other monoclonal antibody conjugated with time-resolved fluorescent microspheres.

[0200] During the binding reaction, the antigen in the sample simultaneously binds to two antibody conjugates. During the capture reaction, the binding reaction mixture reacts with streptavidin coated on a nitrocellulose membrane. After the capture reaction, unbound material is washed away using a reaction buffer.

[0201] Finally, fluorescence signals were collected, and the intensity of the fluorescence signal was positively correlated with the antigen concentration in the sample.

[0202] To more clearly illustrate the technical solution adopted by this utility model, the exemplary operation of the chip and the centrifugation process are described below:

[0203] Step 1) A liquid sac can be pre-stored in the first pool 10, and a sealing strip 20A is provided on the liquid sac. The reaction buffer stored inside can be released by removing the liquid sac; add whole blood sample to the sample pool 10 through the sample application port 101.

[0204] Step 2) Perform the first centrifugation. Under the action of centrifugal force, the buffer enters the third quantitative pool 21 (excess liquid can enter the second waste liquid pool 22); and, under the action of centrifugation, whole blood can be separated, that is, blood is separated into plasma and red blood cells, and plasma enters the first quantitative pool 14.

[0205] Step 3) Perform a second centrifugation. Under the action of centrifugal force, the buffer solution in the third quantitative pool 21 enters the mixing chamber, and the plasma in the first quantitative pool 14 also enters the mixing chamber to achieve full mixing of the buffer solution and plasma. At this time, the buffer solution in the second waste liquid pool 22 will enter the second transition pool 41.

[0206] Step 4) Perform a third centrifugation. Under the action of centrifugal force, the mixture in the mixing chamber continues to enter the fourth quantitative cell 43; while the buffer in the second transition cell 41 moves further to the third transition cell 45; at the same time, the test mixture in the fourth quantitative cell 43 can move through the capillary tube (which has an air hole on the side away from the liquid inlet) and wet the test strip in the detection area, thereby completing the fluorescence reaction with the test strip.

[0207] Step 5) Perform the fourth centrifugation. Under the action of centrifugal force, the buffer solution in the third transition cell 45 will move further to the fourth quantitative cell 43. Under continuous centrifugation, the buffer solution can continue to move into the test strip to clean it, so as to remove impurities that may affect color detection and improve the accuracy of test results.

[0208] It is understandable that the above-mentioned multiple centrifugations can be performed continuously or intermittently in sequence.

[0209] Preferably, the above-mentioned chip can be made into a procalcitonin (PCT) detection kit (microfluidic time-resolved fluorescence immunoassay) product.

[0210] This product is intended for the in vitro quantitative determination of procalcitonin levels in human plasma, serum, or venous whole blood.

[0211] Procalcitonin (PCT) is a glycoprotein with a molecular weight of approximately 12.8 kDa. Under normal physiological conditions, it is produced by thyroid C cells, but during bacterial infections, various tissues and cells also secrete PCT. In clinical diagnosis, PCT levels rise earlier in patients with bacterial sepsis, facilitating early diagnosis and monitoring by physicians. Typically, in systemic infections, serum PCT levels begin to rise within 2–4 hours, peaking within 8–24 hours and remaining elevated for several days or weeks. When levels exceed a certain threshold, the patient should be considered at risk of developing severe sepsis or septic shock. Furthermore, procalcitonin is an important biomarker that can specifically differentiate bacterial infections from inflammatory responses caused by other factors. Viral infections, allergic reactions, autoimmune diseases, and transplant rejection do not cause significant increases in procalcitonin levels, while localized bacterial infections can lead to moderate increases in procalcitonin levels.

[0212] PCT can be used as an adjunct diagnostic tool for bacterial infectious diseases.

[0213] Testing principle:

[0214] The Procalcitonin (PCT) Assay Kit (Microfluidic Time-Resolved Fluorescence Immunoassay) is a centrifugal microfluidic assay tray that uses a double-antibody sandwich method. It employs two monoclonal antibodies that specifically recognize PCT to detect PCT levels in plasma, serum, or whole venous blood. One monoclonal antibody is a biotin conjugate, and the other is a time-resolved fluorescent microsphere conjugate.

[0215] Driven by centrifugal force, the reagent tray sequentially completes the steps of plasma separation, binding reaction, capture reaction, and washing. During the binding reaction, PCT in the sample simultaneously binds to two antibody conjugates. During the capture reaction, the binding reaction mixture reacts with streptavidin coated on a nitrocellulose membrane. After the capture reaction, unbound material is washed away using the reaction solution.

[0216] Finally, the detection line and control line were illuminated with light at a wavelength of 330-365 nm, and fluorescence signals were collected at a wavelength of 605-625 nm. The ratio of the detection line signal to the control line signal was calculated, and the ratio result was positively correlated with the PCT concentration in the sample.

[0217] Main components of the product:

[0218] 1. The test reagent tray is individually packaged in an aluminum foil bag, which contains a desiccant packet.

[0219] 2. Reagent components contained in the product:

[0220] PCT antibody 2 biotin conjugate, PCT antibody 1 microsphere conjugate, liquid capsule (PBS), test strip 1 (streptavidin), and test strip 2 (time-resolved fluorescent microspheres).

[0221] Storage conditions and shelf life

[0222] Store in a cool, dry place at 2–25℃, away from direct sunlight and with an ambient humidity of 30–80%. Shelf life is 12 months.

[0223] Once the aluminum foil bag is opened, the test reagent tray should be used within 20 minutes. After adding the sample, it should be placed in the device for testing immediately.

[0224] Applicable instruments

[0225] Tianjin Mustard Biotechnology Co., Ltd.'s dry fully automated fluorescence immunoassay analyzers JIE-200 and JIE-400.

[0226] Sample Requirements

[0227] Applicable sample types: serum, plasma, and venous whole blood.

[0228] Suitable anticoagulants: lithium heparin and sodium heparin.

[0229] Venous whole blood samples should be tested or separated immediately after collection. Separated serum and plasma can be stably stored at 2–8°C for 24 hours. Samples need to be thoroughly mixed before testing.

[0230] Preferably, when the concentrations of bilirubin, hemoglobin, triglycerides, biotin, human calcitonin, human calcitonin, human calcitonin, human α-CGRP, human β-CGRP, rheumatoid factor, and HAMA in the samples used for testing are below 30 mg / dL, there is no impact on PCT detection.

[0231] Test methods

[0232] Experimental parameters

[0233] The dry-type fully automated fluorescence immunoassay analyzer should be used in an ambient temperature range of 10–30°C. During the test, the reagent tray should be kept at a constant temperature of 37°C, and the test time should be less than 20 minutes.

[0234] Experimental steps

[0235] 1. If the kit is stored at 2–8°C, it can be brought to room temperature before testing.

[0236] 2. Use the barcode scanner provided with the dry fully automated fluorescence immunoassay analyzer to scan the QR code on the aluminum foil bag and read the basic information and preset calibration parameters of the test reagent tray.

[0237] 3. Remove the test reagent tray from the aluminum foil bag, keeping the blue protective film facing upwards, and proceed as follows: Figure 5 In the direction of the middle arrow, pull out the liquid capsule sealing strip (Note: Once the sealing strip is pulled out, the reagent tray should be laid flat to avoid being upside down or tilted at a large angle).

[0238] 4. Use a pipette to add 50 μL of the sample to be tested into the sample dispensing port (see...). Figure 5 Remove the blue protective film from the surface of the test reagent tray.

[0239] 5. Refer to the instruction manual for the dry fully automated fluorescence immunoassay analyzer to place the test reagent tray into the corresponding position on the analyzer tray and complete the subsequent operations.

[0240] calibration

[0241] This product is calibrated by the manufacturer before leaving the factory. The QR code on the packaging provides specific calibration data for each test kit and is read by a dry automated fluorescence immunoassay analyzer before testing.

[0242] result

[0243] The dry fully automated fluorescence immunoassay analyzer will automatically calculate and output the test results after the test is completed. The result unit is ng / mL.

[0244] Reference range

[0245] Referring to the "Expert Consensus on the Emergency Clinical Application of Procalcitonin (PCT)," the reference value for verification was <0.05 ng / mL, based on the test results of 120 normal adults.

[0246] Due to differences in geography, race, gender, and age, it is recommended that each laboratory establish its own reference range.

[0247] Interpretation of test results

[0248] The preferred detection range for this product is 0.05–40 ng / mL. Values ​​below 0.05 ng / mL will be reported as <0.05 ng / mL; values ​​above 40 ng / mL will be reported as >40 ng / mL.

[0249] According to the "Expert Consensus," the following recommendations are made for interpreting PCT results:

[0250] Table 1 - Result Interpretation Table I

[0251]

[0252] Note: PCT levels must be interpreted in conjunction with the clinical situation. Interpretation should be avoided when detached from the patient's specific condition, and the possibility of false positives and false negatives should be considered.

[0253] Clinical significance and management recommendations for PCT levels in patients with respiratory infections, such as

[0254] Below: Table 2 - Result Interpretation Table II

[0255]

[0256] Note: (1) For patients who have already taken antibiotics upon admission, it is recommended to discontinue the antibiotics already in use if PCT < 0.25 ng / mL.

[0257] If the PCT concentration decreases by more than 80% compared to baseline, it is recommended to discontinue antibiotic use. If the decrease is 90%, it is strongly recommended to discontinue antibiotic use.

[0258] Besides infection, the following conditions can also cause elevated PCT levels:

[0259] 1. Prolonged or severe cardiac shock;

[0260] 2. Long-term severe irregular organ perfusion;

[0261] 3. Small cell lung cancer or C-cell tumor of the thyroid medulla;

[0262] 4. Early stage of extensive trauma, surgical procedures, and severe burns;

[0263] 5. Inflammatory cytokine stimulation and release therapy;

[0264] 6. Newborns (within 48 hours of birth).

[0265] Preferred product performance indicators

[0266] Appearance: All components of the reagent kit should be complete, both inner and outer packaging should be intact, liquid reagents should not leak, and labels should be clear.

[0267] Blank limit: not higher than 0.03 ng / mL.

[0268] Limit of detection: not higher than 0.05 ng / mL.

[0269] Limit of quantitation: not higher than 0.15 ng / mL.

[0270] Accuracy: Recovery rate is in the range of 85.0% to 115.0%.

[0271] Linearity: Within the linear range of 0.15–40 ng / mL, the correlation coefficient |r| is not less than 0.9900.

[0272] Repeatability (CV%): not greater than 8.0%.

[0273] Inter-batch variation (CV%): not greater than 11.0%.

[0274] In some embodiments, the fourth, fifth, sixth, and seventh channels are all Class II channels.

[0275] To more clearly illustrate the technical solution adopted by this utility model and its resulting technical effects, see [link to relevant documentation]. Figures 6-15 The technical solution will be described exemplarily below using an alternative expression (and a new set of figure references):

[0276] This centrifugal microfluidic immunoassay reagent tray comprises two layers, the upper and lower layers being tightly fitted together. The upper layer is a cover layer with multiple sets of through-holes for different functions; the lower layer is a channel layer with multiple sets of grooves (or cavities) and channels of different sizes and functions. The grooves may include sample loading cavities, plasma quantification cavities, red blood cell storage cavities, sac placement cavities (buffer solution loading cavities), buffer quantification cavities, buffer transition cavities, reaction cavities, chromatography quantification cavities, test strip placement cavities, control test strip placement cavities, residual liquid storage cavities (or waste liquid cavities), vent structures, and positioning holes (optional). These grooves are primarily connected by various channels (some of which are capillary channels).

[0277] See Figure 10 The diagram shows the structure of the cover plate of a centrifugal microfluidic immunoassay reagent tray. The cover plate 1 has multiple through holes, including one sample inlet 101, one buffer sample inlet 103, and two positioning holes 102.

[0278] in:

[0279] Sample inlet 101, inner diameter: 1 to 5 mm, function: to add the sample into the sample inlet chamber 201 directly below.

[0280] Buffer sample well 103, inner diameter: 1 to 5 mm, function: to add buffer to the liquid bladder placement chamber (buffer sample chamber) 205 directly below (when the liquid bladder is not pre-filled with reaction solution).

[0281] Positioning hole 102, inner diameter: 1 to 30 mm, function: overlaps with positioning hole 208 of channel layer 2 directly below to form a complete positioning hole, used to fix the reagent tray to the centrifuge tray of the matching machine.

[0282] Figure 11 This is a schematic diagram of the channel layer structure of a centrifugal microfluidic immunoassay reagent tray. Channel layer 2 includes a sample loading chamber 201, a plasma quantification chamber 202, a red blood cell storage chamber 203, a liquid sac placement chamber (buffer solution loading chamber) 205, a buffer quantification chamber 206, a buffer transition chamber 207, a reaction chamber 210, a chromatography quantification chamber 211, a test strip placement chamber 213, a control test strip placement chamber 214, a residual liquid storage chamber (or waste liquid chamber) 212, a pore structure 209, a positioning hole 208, and a capillary channel 204. Among these:

[0283] The depth, width, and length of each structure on channel layer 2 can be the same or different, and their dimensions can be, for example, from 50 μm to 50 mm.

[0284] Sample filling chamber 201, volume: 20 to 500 mm³ 3 Function: Stores samples, which can be common biological samples such as whole blood, serum, plasma, and urine.

[0285] Plasma quantification chamber 202, volume: 5 to 500 mm 3 Function: to separate and quantify plasma.

[0286] Red blood cell storage cavity 203, volume: 20 to 500 mm 3 Function: If the sample is whole blood, it can hold red blood cells separated by centrifugation. However, it is understood that the sample is not limited to whole blood; as long as it contains cells, other particulate matter, or no particulate matter, the red blood cell storage chamber 203 can also be used to hold such cells, other particulate matter, or particulate-free liquids. Therefore, names such as "red blood cell storage chamber" are used in this document only for ease of description and understanding, and not as a clear limitation on the purpose of the chamber.

[0287] Residual liquid storage chamber (or waste liquid chamber) 212, volume: 20 to 2000 mm³ 3 Function: to store waste liquids, such as excess samples, excess liquid after reaction, excess buffer solution, etc.

[0288] Liquid-filled bladder placement chamber (buffered sample chamber) 205, volume: 10 to 2000 mm³ 3 Function: This tray contains a pre-filled sac (3) for storing buffer solutions. Before use, the sac should be opened by puncturing or tearing to release the buffer solution. The buffer solution can be common buffers, such as PB, Tris, HEPES, TAPS, MOPS, MES buffers at concentrations from 1mM to 2000mM. The buffer solution may contain proteins such as BSA, casein, gelatin, and IgG, surfactants such as Tween and Triton, and polymers such as sugars, PEG, PVA, and PVP. The buffer solution in this reagent tray has both dilution and washing functions. See the schematic diagram of the sac structure. Figure 12 The liquid bladder in this example consists of a sealing strip 301 and a liquid bladder shell 302. The liquid bladder is released by tearing (pulling out the sealing strip 301). The specific shape and size of the liquid bladder can be used in conjunction with the specific structure of the liquid bladder placement cavity (buffer solution plus sample cavity) 205. The specific shape, material, and puncture method of the liquid bladder are not limited to this example.

[0289] Buffer metering chamber 206, volume: 5 to 500 mm³ 3 Function: Quantitative buffer. The quantified buffer is used for sample dilution.

[0290] Buffer transition chamber 207, volume: 5 to 500 mm³ 3 Function: Used for buffer storage and transition. The buffer here is used for subsequent washing.

[0291] Reaction chamber 210, volume: 10 to 1000 mm3 Function: This area provides a site for binding reactions and can be pre-loaded with drying reagents, such as antibody-conjugated labels dried by a specific method (e.g., lyophilization, ultrasonic spray drying). Examples of labels include commonly used immunochromatographic markers such as biotin, colloidal gold, alkaline phosphatase, fluorescent dyes, fluorescent microspheres, and acridinium esters. One or more drying reagents can be used. If no drying reagent is pre-loaded here, this area can provide a buffer solution for sample dilution.

[0292] Quantitative chromatography chamber 211, volume: 5 to 500 mm³ 3 Function: To quantitatively supply liquid from reaction chamber 210 for the capture reaction of the test strip. Alternatively, to quantitatively supply buffer from buffer transition chamber 207 for washing the test strip after the capture reaction.

[0293] Test strip placement cavity 213, dimensions: width 1 to 10 mm, length 10-100 mm, depth 1 to 5 mm. Function: To place test strip 4. See the schematic diagram of the test strip structure. Figure 13 It typically consists of a base plate 401, a reaction pad 402, and an absorbent pad 403, but may also include other structures (such as a sample pad). The reaction pad 402 is coated with biomaterials with capture functions, such as streptavidin that can bind to biotin, or antibodies that can bind to antigens.

[0294] The control test strip is placed in cavity 214. Dimensions: width 2-10 mm, length 3-60 mm, depth 1-5 mm. Function: To hold control test strip 5. See the schematic diagram of the control test strip structure. Figure 14 It typically consists of a substrate 501 and a biofilm 502. The biofilm 502 is marked with markers, such as colloidal gold, fluorescent dyes, or fluorescent microspheres; these markers on the control strip are the control lines (C line, control line). Traditionally, both the control line and the test line are on the same test strip, requiring a reaction to obtain the control line result, thus affecting the repeatability of the test results. This method uses an independent control line, which does not participate in the reaction, resulting in good repeatability.

[0295] Figure 14 The diagram shows the structure of the test strip, with a top view and a bottom side view.

[0296] Pore ​​structure 209, size not limited, function: to maintain air pressure balance and ensure smooth liquid flow.

[0297] Positioning hole 208, inner diameter: 1 to 30 mm, function: used to fix the reagent tray to the centrifuge tray of the matching machine.

[0298] Capillary channel 204, with an inner diameter of 50μm to 500μm, functions to control liquid flow through a siphon principle. When centrifugation continues, because the liquid level is below the highest point of the capillary channel, the centrifugal force is greater than the capillary action, preventing the liquid from passing through and causing it to remain in the upstream chamber. When centrifugation stops, the liquid fills the capillary channel due to capillary action. Upon subsequent centrifugation, the liquid enters the downstream chamber. The capillary channel 204 can be modified, for example, by applying a hydrophilic treatment to the surface layer of the channel through which the liquid passes, to facilitate smooth liquid flow within the channel.

[0299] A schematic diagram of the overall structure of this centrifugal microfluidic immunoassay reagent tray is shown below. Figure 15 .

[0300] This centrifugal microfluidic immunoassay reagent tray can be round, square, or polygonal. The reagent tray can exist individually or multiple trays can be combined into a single unit (e.g., [missing information - likely a combination of two trays]). Figure 6 The six sector-shaped reagent trays are combined to form a complete circular reagent tray. Taking the complete circular reagent tray as an example, the diameter can be from 3cm to 30cm.

[0301] The centrifugal microfluidic immunoassay reagent tray can be made of plastic materials such as cyclic olefin copolymer (COC), polymethyl methacrylate (PMMA), and polystyrene (PS). The cover layer 1 can be colorless and transparent or opaque (e.g., black), with a thickness of 0.1 to 1 mm; the channel layer 2 can be transparent or opaque (e.g., black), with a thickness of 0.5 to 20 mm.

[0302] This centrifugal microfluidic immunoassay reagent tray can be manufactured by machining, etching, mold casting, etc., with the preferred method being injection molding.

[0303] The cover layer 1 and channel layer 2 of this centrifugal microfluidic immunoassay reagent tray can be connected by adhesive bonding, ultrasonic welding or laser welding.

[0304] When used to test whole blood samples, a portion of the dried reagent for testing can be pre-loaded into the reaction chamber 210, the test strip 4 into the test strip placement chamber 213, the control test strip 5 into the test strip placement chamber 214, and the liquid capsule 3 into the liquid capsule placement chamber (buffer solution addition chamber) 205. The whole blood sample is added to the sample addition chamber 201 through the sample port 101. Simultaneously, the liquid capsule is ruptured, and under centrifugal force, the whole blood sample flows sequentially to the plasma quantification chamber 202 and the red blood cell storage chamber 203. Under centrifugal force, the whole blood sample is separated into a plasma layer and a blood cell layer (mainly red blood cells), with the blood cell layer located in the red blood cell storage chamber 204 and the plasma layer located in the plasma separation chamber 203 (part of the plasma layer may also be located in the red blood cell storage chamber 204). Simultaneously, the buffer solution in the liquid capsule flows sequentially to the buffer quantification chamber 206 and the buffer transition chamber 207. Plasma in plasma quantification chamber 202 and buffer solution in buffer quantification chamber 206 then enter reaction chamber 210 to dissolve the pre-filled dried reagent, followed by a binding reaction in reaction chamber 210. After the reaction, the reaction complex enters chromatography quantification chamber 211, contacts the test strip 4, and undergoes chromatography, thereby capturing the biomaterial with capture function on it. After the reaction, buffer solution in buffer transition chamber 207 enters chromatography quantification chamber 211, contacts the test strip 4, and undergoes chromatography, achieving the cleaning function.

[0305] This embodiment provides an interleukin-6 detection reagent tray that can be used for clinical medical diagnosis. (Reference) Figures 10 to 15 The reagent tray is made of transparent PMMA and is fan-shaped. The cover layer 1 is 0.6 mm thick and the channel layer 2 is 3 mm thick. Both the cover layer 1 and the channel layer 2 are made by injection molding. After injection molding, the capillary channels 204 on the channel layer 2 are hydrophilically treated with 1% Tween 80.

[0306] The inner diameter of sample dispensing port 101 and buffer solution dispensing well 103 is 2 mm, and the inner diameter of positioning holes 102 and 208 is 4 mm. The volume of sample dispensing cavity 201 is 200 mm³. 3 The volume of the plasma quantification chamber 202 is 5 mm. 3 The volume of the red blood cell storage cavity 203 is 30 mm. 3 The volume of the 205-inch fluid reservoir (buffered sample chamber) is 800 mm². 3 The volume of the buffer quantification chamber 206 is 40 mm. 3 The volume of the buffer transition chamber 207 is 100-120 mm. 3 The volume of reaction chamber 210 is 100 mm. 3The reaction chamber 210 is pre-loaded with biotin-conjugated IL-6 antibody 2706 (Medix) and fluorescent microsphere-conjugated IL-6 antibody 2704 (Medix). The pre-loaded reagents are dried by spotting and drying at 37°C for 2 hours.

[0307] The volume of the chromatography quantitative chamber 211 is 13 mm. 3 The test strip placement cavity 213 has dimensions of 3mm width, 35mm length, and 1mm depth. The test strip consists of a base plate 401, a reaction pad 402, and an absorbent pad 403. The base plate 401 is made of PVC material DB-7, the absorbent pad 403 is made of pure cotton pulp filter paper H5072, and the reaction pad 402 is made of nitrocellulose membrane AE99 (Whatman). Streptavidin SA101 (Merck) is etched onto the reaction pad 402.

[0308] The dimensions of the control test strip in cavity 214 are: width: 3mm, length: 6mm, depth: 1mm. The control test strip consists of a base plate 501 and a biofilm 502. The base plate 501 is made of PVC material DB-7, and the biofilm 502 is a nitrocellulose membrane AE99 (Whatman). Fluorescent microspheres FCEU002 (Bangs) are drawn on the biofilm 502.

[0309] Preferably, the inner diameter of all capillary channels 204 is 300 μm.

[0310] After the reagent tray was prepared, it was used in conjunction with a dry-type fully automated fluorescence immunoassay analyzer to perform repeatability testing and methodological comparison analysis on IL-6 samples. The results are as follows:

[0311] Figure 16 The repeatability results of repeated tests were performed on the method of this invention and the conventional chromatography method. Figure 17 The methodological comparison results (vs. Roche's electrochemiluminescence method);

[0312] Figure 16 In this context, AV represents the mean, SD represents the standard value, and CV represents the coefficient of variation for repeatability.

[0313] in, Figure 17 The x-axis represents the results obtained using the Roche electrochemiluminescence method, and the y-axis represents the results obtained using the chip of this invention. The sample size n is 23. The correlation coefficient R is calculated based on the results from the two methods. 2 It is 0.9919;

[0314] Results analysis: Traditional chromatography methods have repeatability CVs above 10%, while the repeatability CVs of this method are all <5%. Methodological comparisons were performed with Roche's (electrochemiluminescence) method, and R... 2A value >0.99 indicates good consistency. Therefore, this method performs better.

[0315] Example 2

[0316] The centrifugation control program / method for the chip provided in Example 1 includes the following steps:

[0317] 1. 2000-8000 rpm, 5-300 s. Purpose: ① Whole blood separation and plasma quantification; ② Buffer solution quantification.

[0318] 2. Remain still for 1-40 seconds. Function: To open the capillary switch.

[0319] 3. ①2000-6000rpm, 1-10s; ②1000-4000rpm, 1-10s.

[0320] Repeat steps ① to ② 5-200 times. Function: To carry out a binding reaction.

[0321] 4. Remain still for 1-40 seconds. Function: To open the capillary switch.

[0322] 5. 500-8000 rpm, 1-40 s. Function: To measure the reaction mixture.

[0323] 6. Remain still for 30-500 seconds. Functions: ① To open the capillary switch; ② To initiate a capture reaction.

[0324] 7. 500-8000 rpm, 1-40 s. Purpose: Quantification of buffer solution.

[0325] 8. Remain still for 30-500 seconds. Purpose: Washing.

[0326] References

[0327] 1. Expert consensus group on the emergency clinical application of procalcitonin. Expert consensus on the emergency clinical application of procalcitonin (PCT) [J]. Chinese Journal of Emergency Medicine, 2021, 21(9): 944-951.

[0328] 2. Expert consensus on the clinical significance of infection-related biomarkers, Chinese Medical Education Association Infectious Diseases Committee [J]. Chinese Journal of Tuberculosis and Respiratory Diseases, 2017, 40(4): 243-257.

[0329] 3.Whang KT,Vath SD,Becker KL,et al.Procalcitonin and proinflammatoryeytokine interactions in sepsis[J].Shock,2000,14(1):73-78.

[0330] 4.Chalupa P,Beran O,Herwald H,et al.Evaluation of potentialbiomarkers for the discrimination of bacterial and viral infections[J].Infection,2011,39(5):411-417.

[0331] 5.Christ-Crain M,Jaccard-Stolz D,Bingisser R,et a1.Effect ofprocalcitonin-guided treatment on antibiotic use and outcome in lowerrespiratory tract infections:cluster-randomised,single-blinded interventiontria[J].Lancet,2004,363(9409):600-607.

[0332] 6.Schuetz P,Christ-Crain M,Thomann R,et al.Effect of procalcitonin-based guidelines vs standard guidelines on antibiotic use in lowerrespiratory tract infections:the ProHOSP randomized controlled trial[J].JAMA,2009,302(10):1059-1066.

[0333] 7.Schuetz P,Christ-Crain M,Wolbers M,et a1.Procalcitonin guidedantibiotic therapy and hospitalization in patients with lower respiratorytract infections:a prospective,multicenter,randomized controlled trial[J].BMCHealth Serv Res,2007,7:102.

[0334] It should be noted that, in this document, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes that element.

[0335] Through the above description of the embodiments, those skilled in the art can clearly understand that the methods of the above embodiments can be implemented by means of software plus necessary general-purpose hardware platforms. Of course, they can also be implemented by hardware, but in many cases the former is a better implementation method. Based on this understanding, the technical solution of the present invention, or the part that contributes to the prior art, can be embodied in the form of a software product. This computer software product is stored in a storage medium (such as ROM / RAM, magnetic disk, optical disk) and includes several instructions to cause a computer terminal (which may be a mobile phone, computer, server, or network device, etc.) to execute the methods described in the various embodiments of the present invention.

[0336] The embodiments of the present invention have been described above with reference to the accompanying drawings. However, the present invention is not limited to the specific embodiments described above. The specific embodiments described above are merely illustrative and not restrictive. Those skilled in the art can make many other forms under the guidance of the present invention without departing from the spirit and scope of the claims. All of these forms are within the protection scope of the present invention.

Claims

1. A chip, characterized in that, The chip is sequentially provided with a first centrifugation area, a second centrifugation area, a third centrifugation area, and a detection area; The first centrifugation zone is equipped with: A sample addition pool (10) is provided with a sample addition port (101) for adding samples; First pool (20), the first pool is used to contain buffer solution; The second centrifuge zone is equipped with: The first quantitative cell (14) is connected to the sample dispensing cell (10) through at least one Class I channel; A third metering pool (21) is connected to the first pool (20) via at least one of the Class I channels; The third centrifugation zone is equipped with: The mixing chamber (30) is connected to the first metering cell (14) and the third metering cell (21) through a Class II channel; The sample in the first quantitative cell (14) and the buffer solution in the third quantitative cell (21) can enter the mixing chamber synchronously under the action of centrifugal force and be mixed in the mixing chamber to form a mixture; The detection area is provided with a detection position, which is connected to the mixing chamber and is used to place the reactants. The mixture continues to enter the detection position under the action of centrifugal force.

2. The chip according to claim 1, characterized in that, A first transition pool (17) is also provided between the sample loading pool (10) and the first quantitative pool (14). The first transition pool (17) is connected to the sample loading pool (10) through the Class I channel. And / or, the chip further includes: a first waste liquid pool (15), a first side of the first quantitative pool (14) being connected to the first waste liquid pool (15) via at least one of the Class I channels, the first waste liquid pool (15) being used to contain excess sample overflowing from the first quantitative pool (14) so ​​that the first quantitative pool (14) can quantitatively regulate the sample entering the mixing chamber (30).

3. The chip according to claim 1, characterized in that, Also includes: The container (16) is connected to the second side of the first quantitative container (14). The second side is provided with a first groove (141) and a second groove (142) that are connected. The first groove (141) is connected to the mixing chamber through the type II channel, and the depth of the first groove (141) is greater than that of the second groove (142) so as to facilitate the drainage and collection of the sample. The container (16) is used to collect the precipitate obtained by centrifugation.

4. The chip according to claim 1, characterized in that, The third quantitative cell (21) includes: The first sub-pool (211) and the second sub-pool (212) are connected; and the bottom surface of the first sub-pool is higher than the bottom surface of the second sub-pool. The chip further includes: a second waste liquid tank (22), wherein the third quantitative tank (21) is connected to the second waste liquid tank (22), and wherein, The second waste liquid pool (22) includes a connecting pool (222) and a storage pool (221), the inlet of which is located on the first side of the first sub-pool (211); the second waste liquid pool (22) is used to contain excess buffer solution overflowing from the third quantitative pool (21).

5. The chip according to claim 4, characterized in that, The chip also includes: The second transition pool (41) is connected to the second waste liquid pool (22) through at least one of the Class II channels; The third transition pool (45) is connected to the second transition pool (41) through at least one of the Class II channels, and the third transition pool is connected to the detection position.

6. The chip according to claim 5, characterized in that, Also includes: The fourth quantitative cell (43) is connected to the third transition cell (45) through at least one of the type II channels; the mixing chamber is connected to the fourth quantitative cell (43) through the type II channel; the fourth quantitative cell (43) is also connected to the detection position through a type I channel.

7. The chip according to claim 6, characterized in that, The second waste liquid tank (22), the second transition tank (41), the third transition tank (45) and the fourth quantitative tank (43) are arranged in a direction that moves away from the centrifuge center in sequence.

8. The chip according to claim 6, characterized in that, The fourth metering chamber (43) includes a first connecting chamber (433), a second connecting chamber (431), and a metering chamber (432); wherein the first connecting chamber (433) is connected to the third transition chamber (45) and the mixing chamber respectively; the inlet end of the metering chamber (432) is connected to the outlet end of the first connecting chamber (433), and the inlet end of the second connecting chamber (431) is connected to the inlet end of the metering chamber (432).

9. The chip according to any one of claims 1-8, characterized in that, The type II channel is a U-shaped capillary channel, and the U-shaped opening of the U-shaped capillary channel is oriented away from the centrifugal center.

10. The chip according to claim 1, characterized in that, The chip is provided with at least one air hole (35) connected to the mixing chamber for maintaining air pressure balance; and / or, the chip is provided with a control area for setting a control substance.

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