A microfluidic chip adaptable to fluorescent quantitative PCR instrument

Abstract

The present invention discloses a microfluidic chip that can be adapted to a fluorescent quantitative PCR instrument, which belongs to the field of molecular diagnosis technology and is used to solve the problems of uneven sample loading and complicated procedures in the prior art when testing large quantities of samples. The microfluidic chip includes a chip body with a plurality of visualized reaction pools, the shape of the chip body is adapted to the PCR instrument, and at least one bent microfluidic channel is provided in the chip body, and the lower convex parts of the microfluidic channel are connected to the reaction pools. The capillary bending area at the upper convex part of the microfluidic can prevent the liquid in the microfluidic channel from flowing due to centrifugal acceleration during centrifugal separation, so that the sample loading liquid flowing into the reaction pool is more uniform, thereby solving the problem of uneven sample loading of the sample loading liquid and improving the use effect.

Description

Technical Field

[0001] The present invention relates to the technical field of molecular diagnosis, and in particular to a microfluidic chip adaptable to a fluorescence quantitative PCR instrument. Background Art

[0002] Fluorescence quantitative PCR technology is a very common quantitative detection technology in the field of molecular diagnosis. It is achieved by mixing reagents and samples in an 8-well, 96-well plate, and then placing them in a dedicated fluorescence quantitative PCR instrument for temperature-controlled reaction. As the reaction proceeds, a specific fluorescence signal is generated, and the instrument collects the signal to achieve the diagnostic function.

[0003] Common detection indicators may involve multiple ones, and a detection sample needs to be repeatedly added to multiple reaction units. Currently, the common reaction containers that can be matched with fluorescent quantitative PCR instruments are mainly PCR tubes, eight-well strips and 96-well plates. In the case where a single sample needs to be reacted in multiple reaction units, manual repeated sample addition operations are required, which increases the repetitive work of operators. This situation is more common, especially in medical institutions and testing institutions.

[0004] In the prior art, when adding the sample liquid to multiple reaction units at one time, it is necessary to add an isolation liquid to isolate the flow of the sample liquid in the sample channel, which makes the process cumbersome. If the isolation liquid is not added, it is easy to cause uneven liquid separation during liquid separation, affecting the final detection effect. Summary of the invention

[0005] In view of the above problems, the present invention proposes a microfluidic chip that can be adapted to a fluorescent quantitative PCR instrument, which is used to solve the problems of uneven sample addition and complicated procedures in the prior art when testing large quantities of samples.

[0006] The technical solution of the present invention is as follows: the microfluidic chip comprises a chip body 1 having a plurality of visualized reaction pools 5, the shape of the chip body 1 being adapted to a PCR instrument, at least one curved microfluidic channel 4 being provided in the chip body 1, and the lower convex parts of the microfluidic channels 4 being connected to the reaction pools 5.

[0007] The number of the microfluidic channels 4 on the upper body of the chip body 1 is 1 to 12. When the number of the microfluidic channels 4 is greater than 1, all the microfluidic channels 4 are arranged in parallel.

[0008] Further, the meandering microfluidic channel 4 includes a plurality of upper convex channels 3 and lower convex channels 2 that are alternately arranged and sequentially connected, and the upper convex channels 3 and lower convex channels 2 that are sequentially connected can constitute a meandering microfluidic channel 4; the lower convex channels 2 correspond to the reaction pools 5 one by one, and the bottoms of the plurality of lower convex channels 2 are respectively connected to the plurality of reaction pools 5;

[0009] The upper convex channel 3 is in an inverted V shape or an inverted U shape; the lower convex channel 2 is in a V shape or a U shape.

[0010] For example 1, the upper convex channel 3 is in an inverted V shape, the lower convex channel 2 is in a V shape, and the reaction pool 5 is connected to the corner of the V-shaped channel.

[0011] In the second example, the upper convex channel 3 is in an inverted V shape, the lower convex channel 2 is in a U shape, and the reaction pool 5 is connected to the bottom of the U-shaped channel.

[0012] Furthermore, the upper convex channel 3 includes an upper double-arm area 301 and an upper bending area arranged at the top thereof, the upper double-arm area 301 includes two symmetrically arranged upper channels, and the two ends of the upper bending area are respectively connected to the two upper channels;

[0013] The lower convex channel 2 includes a lower double-arm area 201 and a lower bending area at the bottom thereof, wherein the lower double-arm area 201 includes two symmetrically arranged lower channels, and the two ends of the lower bending area are respectively connected to the two lower channels;

[0014] The bottom of the upper channel in the upper convex channel 3 and the top of the lower channel in the lower convex channel 2 are connected to each other, so that the adjacent upper convex channels 3 and lower convex channels 2 remain connected.

[0015] The upper bending zone is a capillary bending zone 302, and the bending zone of the lower convex channel 2 is a transition bending zone 202. The cross-section of the capillary bending zone 302 is smaller than the cross-section of the transition bending zone 202. The lower convex channel 2 also includes a capillary transition zone 203 connected to the transition bending zone 202. The lower convex channel 2 is connected to the reaction pool 5 through the capillary transition zone 203.

[0016] Furthermore, a liquid separation unit can be formed between any lower convex channel 2 of the microfluidic channel 4 and the tops of the two adjacent upper convex channels 3, each liquid separation unit is connected to the reaction pool 5, and the number of liquid separation units of each microfluidic channel 4 is at least 2.

[0017] Furthermore, the height difference between the upper convex part and the lower convex part of the bent microfluidic channel 4 is H value, and the range of H value is: 0.1mm to 5mm.

[0018] Furthermore, the microfluidic chip further comprises a cover sheet 8 seamlessly connected to the chip body 1 , and the shape of the cover sheet 8 is adapted to the shape of the chip body 1 .

[0019] Furthermore, at least a portion of the cover sheet or the chip body is made of a colorless and transparent material for visualizing the reaction pool.

[0020] Furthermore, the volume of the reaction pool is 2uL to 1000uL.

[0021] The beneficial effects of the present invention are as follows: the present case comprises a chip body having a plurality of visualized reaction pools, the shape of the chip body being adapted to a PCR instrument, at least one curved microfluidic channel being arranged in the chip body, and the lower convex parts of the curved microfluidic channels being connected to the reaction pools.

[0022] Its function is: through the curved microfluidic channel, the capillary bending area at the upper convex part of the microfluidic can prevent the liquid in the microfluidic channel from flowing due to centrifugal acceleration during centrifugal separation, so that the sample liquid flowing into the reaction pool is more uniform, thereby solving the problem of uneven sample liquid loading and improving the use effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 It is a schematic diagram of the three-dimensional structure of the chip body;

[0024] Figure 2 It is a schematic diagram of the structure of the chip body;

[0025] Figure 3 It is a structural schematic diagram of the liquid separation unit;

[0026] Figure 4 for Figure 3 A local enlarged schematic diagram of the middle A;

[0027] Figure 5 is a schematic diagram of the structure of the cover plate;

[0028] Figure 6 It is a schematic diagram of the exploded three-dimensional structure of the microfluidic chip;

[0029] Figure 7 A schematic diagram of the structure of an adapter board that matches the chip body.

[0030] Reference numerals:

[0031] 1-chip body, 2-lower convex channel, 201-lower double-arm area, 202-transition bending area, 203-capillary transition area, 3-upper convex channel, 301-upper double-arm area, 302-capillary bending area, 4-microfluidic channel, 5-reaction pool, 6-sample addition hole, 7-exhaust hole, 8-cover sheet, 9-adapter plate. DETAILED DESCRIPTION

[0032] In order to clearly illustrate the technical features of this patent, this patent is elaborated in detail below through a specific implementation method and in combination with its accompanying drawings.

[0033] In the description of the present utility model, it should be noted that the terms "center", "up", "down", "left", "right", "vertical", "longitudinal", "lateral", "horizontal", "inside", "outside", "front", "back", "top", "bottom", etc. indicate directions or positional relationships based on the directions or positional relationships shown in the accompanying drawings, or are the directions or positional relationships in which the utility model product is usually placed when in use. They are only for the convenience of describing the present utility model and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific direction, be constructed and operated in a specific direction, and therefore cannot be understood as a limitation on the present utility model.

[0034] In the description of the present invention, it is also necessary to explain that, unless otherwise clearly specified and limited, the terms "set", "open", "installed", "connected", and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0035] The present invention is described in detail below with reference to the accompanying drawings and in combination with embodiments:

[0036] like Figures 1 to 7 The microfluidic chip includes a chip body 1 having a plurality of visualized reaction pools 5. The shape of the chip body 1 is adapted to a PCR instrument. At least one curved microfluidic channel 4 is provided in the chip body 1. The lower convex parts of the curved microfluidic channels 4 are connected to the reaction pools 5.

[0037] The shape of the present microfluidic chip can match the adapter plate 9 inside the common fluorescent quantitative PCR instrument in the existing market, and it is also possible to design an adapter plate 9 that matches the present microfluidic chip because of the adjustment of the shape of the design of the present microfluidic chip. The shape of the adapter plate 9 is adapted to the holding slot in the fluorescent quantitative PCR instrument, and the adapter plate 9 is fixedly connected with a plurality of receiving seats distributed in an array, and the receiving seat is provided with a receiving slot adapted to the bottom of the outer wall of the reaction pool in the microfluidic chip. When in use, the microfluidic chip can be vertically placed in the adapter plate, and the bottom of the outer wall of each reaction pool is inserted into each receiving seat, and then the adapter plate is placed in the holding slot in the fluorescent quantitative PCR instrument as a whole, and finally the fluorescent quantitative PCR instrument is turned on for temperature control reaction, which effectively overcomes the inconvenience of use, complicated operation, slow operation speed, and a series of defects when the prior art 8-row and 96-well plates are used for fluorescent quantitative PCR instruments, fills the gap in the prior art, and also solves the problem of uneven sample addition of the sample addition liquid, and has a good practical use effect.

[0038] In this embodiment, Figure 7 The shape of the microfluidic chip matches the existing adapter plate 9.

[0039] Specifically, the number of the microfluidic channels 4 on the upper body of the chip body 1 is 1 to 12. When the number of the microfluidic channels 4 is greater than 1, all the microfluidic channels 4 are arranged in parallel.

[0040] In this embodiment, the number of the microfluidic channel 4 is one.

[0041] Specifically, the microfluidic channels 4 each include a plurality of upper convex channels 3 and lower convex channels 2 that are connected in sequence, and the upper convex channels 3 and lower convex channels 2 that are connected in sequence can form a bent microfluidic channel 4;

[0042] The shape of the upper convex channel 3 includes any one of an inverted V shape or an inverted U shape or a combination thereof;

[0043] The shape of the lower convex channel 2 includes any one of a V shape and a U shape or a combination thereof.

[0044] In this embodiment, Figure 1 and Figure 2 The upper convex channel 3 is inverted V-shaped, and the top of the inverted V-shaped channel has a folded corner. The folded corner can limit the flow of the sample when the microfluidic chip centrifuges the sample liquid to separate the sample, so that the separation is uniform, thereby improving the use effect.

[0045] The shape of the lower convex channel 2 is V-shaped, and the reaction pool 5 is connected to the corner of the V-shaped channel, so that the angle range of the channel connecting the V-shaped channel and the reaction pool 5 is 90 degrees to 180 degrees, so that the sample liquid can enter the reaction pool 5 more easily during centrifugal separation, thereby improving the separation efficiency.

[0046] Specifically, the upper convex channel 3 is in an inverted V shape, the lower convex channel 2 is in a V shape, and the reaction pool 5 is connected to the corner of the V-shaped channel.

[0047] Specifically, Figure 3 and Figure 4 The upper convex channel 3 includes an upper double-arm area 301 and an upper bending area, and the two ends of the upper bending area are respectively connected to the upper double-arm area 301. The lower convex channel 2 includes a lower double-arm area 201 and a lower bending area, one end of the lower double-arm area 201 is connected to the upper double-arm area 301, and the other end of the lower double-arm area 201 is connected to the lower bending area.

[0048] Specifically, the upper bending zone is a capillary bending zone 302, the bending zone of the lower convex channel 2 is a transition bending zone 202, the cross-sectional area of ​​the capillary bending zone 302 is smaller than the cross-sectional area of ​​the transition bending zone 202, the lower convex channel 2 further includes a capillary transition zone 203 connected to the transition bending zone 202, and the lower convex channel 2 is connected to the reaction pool 5 through the capillary transition zone 203. The cross-sectional area of ​​the capillary bending zone 302 is much smaller than the cross-sectional area of ​​the transition bending zone 202.

[0049] The transition bending zone 202 can temporarily store part of the sample liquid before the chip is centrifuged for separation, and the capillary bending zone 302 can isolate or significantly reduce the flow of the sample liquid when the chip is centrifuged, so that the sample liquid can evenly pass through the capillary transition zone 203 into the reaction pool 5 for reaction. At the same time, the lower convex channel 2 is connected to the reaction pool 5 through the capillary transition zone 203, which can have a capillary siphon effect during centrifugation, thereby making it easier for the sample liquid in the transition bending zone 202 to enter the reaction pool 5.

[0050] Specifically, a liquid separation unit can be formed between any lower convex channel 2 of the microfluidic channel 4 and the tops of the two adjacent upper convex channels 3, each liquid separation unit is connected to the reaction pool 5, and the number of liquid separation units of each microfluidic channel 4 is at least 2.

[0051] In this embodiment, Figures 1 to 7 , the number of liquid separation units and reaction pools 5 is 8.

[0052] Specifically, the height difference between the upper convex part and the lower convex part of the curved microfluidic channel 4 is H value, and the range of H value is: 0.1mm to 5mm. H value can be: 0.1mm, 0.5mm, 1mm, 1.5mm, 2mm, 2.5mm, 3mm, 3.5mm, 4mm, 4.5mm, 5mm.

[0053] Specifically, it also includes a cover sheet 8 connected to the chip body 1 without a gap, and the shape of the cover sheet 8 is adapted to the shape of the chip body 1.

[0054] The seamless connection can be achieved by ultrasonic welding, chemical bonding, double-sided adhesive bonding, etc.

[0055] In this embodiment, a sample adding unit connected to the microfluidic channel 4 is also included, and the sample adding unit includes a sample adding hole 6 and a vent hole 7 respectively connected to the microfluidic channel 4. The sample adding hole 6 and the vent hole 7 are respectively arranged at both ends of the cover sheet 8 and are respectively connected to both ends of the microfluidic channel 4.

[0056] Specifically, at least a portion of the cover sheet 8 or the chip body 1 is made of a colorless and transparent material for visualizing the reaction pool 5 .

[0057] In this embodiment, the cover sheet 8 or the chip may be made of colorless and transparent glass or plastic, so as not to affect the collection of fluorescent signals.

[0058] Specifically, the volume of the reaction pool 5 is 2uL to 1000uL. The volume of the reaction pool 5 can be 2uL, 10uL, 50uL, 100uL, 200uL, 500uL, or 1000uL.

[0059] The working process of this embodiment is that different reaction substances are pre-added to the reaction pool 5 when a single sample is tested, which may be primers, reaction systems, etc. The state of the pre-added substances may be dried primers or reaction systems, or lyophilized powder or gel substances with protective agents. The test personnel can add the test object or the mixed reaction system to the microfluidic channel 4 of the microfluidic chip through the sample addition hole 6 through a single sample addition operation. Through centrifugation, the liquid in the microfluidic channel 4 will be centrifuged into the reaction pool 5 below, and the sample addition operation can be completed. After completion, it can be directly put into the fluorescent quantitative PCR instrument for subsequent detection operations.

[0060] Since the shape of the microfluidic channel 4 in this embodiment is bent, and the upper convex channel 3 and the lower convex channel 2 of the bent microfluidic channel 4 are inverted V-shaped and V-shaped respectively, and the top corner of the upper convex channel 3 has a capillary bending area 302, and the transition bending area 202 of the lower convex channel 2 is connected to the reaction pool 5 through the capillary transition area 203, it can make it difficult for the analyte to flow to other liquid separation units during centrifugal separation, and it is easy to enter the reaction pool 5 for reaction, thereby avoiding uneven liquid separation or the need to add an isolation liquid to make the process cumbersome, and improving work efficiency.

[0061] It is understood that the above embodiments are merely exemplary embodiments used to illustrate the principles of the present invention, but the present invention is not limited thereto. For those skilled in the art, various modifications and improvements can be made without departing from the spirit and essence of the present invention, and these modifications and improvements are also considered to be within the scope of protection of the present invention.

Claims

1. A microfluidic chip that can be adapted to a fluorescent quantitative PCR instrument, characterized in that: The microfluidic chip comprises a chip body (1) having a plurality of visible reaction pools (5), the shape of the chip body (1) being compatible with a PCR instrument, at least one curved microfluidic channel (4) being provided in the chip body (1), and the lower convex parts of the microfluidic channels (4) being connected to the reaction pools (5); The meandering microfluidic channel (4) comprises a plurality of upper convex channels (3) and lower convex channels (2) which are alternately arranged and sequentially connected; The upper convex channel (3) comprises an upper double-arm region (301) and an upper bending region arranged at the top thereof, the upper double-arm region (301) comprises two symmetrically arranged upper channels, and the two ends of the upper bending region are respectively connected to the two upper channels; The lower convex channel (2) comprises a lower double-arm region (201) and a lower bending region arranged at the bottom thereof, the lower double-arm region (201) comprises two symmetrically arranged lower channels, and the two ends of the lower bending region are respectively connected to the two lower channels; The bottom of the upper channel in the upper convex channel (3) and the top of the lower channel in the lower convex channel (2) are connected to each other, so that the adjacent upper convex channel (3) and lower convex channel (2) remain connected; The upper bending zone is a capillary bending zone (302), the bending zone of the lower convex channel (2) is a transition bending zone (202), the cross section of the capillary bending zone (302) is smaller than the cross section of the transition bending zone (202), the lower convex channel (2) further comprises a capillary transition zone (203) connected to the transition bending zone (202), and the lower convex channel (2) is connected to the reaction pool (5) via the capillary transition zone (203); The number of microfluidic channels (4) on the upper body of the chip main body (1) is 1 to 12. When the number of microfluidic channels (4) is greater than 1, all microfluidic channels (4) are arranged in parallel.

2. A microfluidic chip adaptable to a fluorescent quantitative PCR instrument according to claim 1, characterized in that: The meandering microfluidic channel (4) comprises a plurality of upper convex channels (3) and lower convex channels (2) which are alternately arranged and sequentially connected, the lower convex channels (2) corresponding to the reaction pools (5) one by one, and the bottoms of the plurality of lower convex channels (2) are respectively connected to the plurality of reaction pools (5); The shape of the upper convex channel (3) is an inverted V shape or an inverted U shape; the shape of the lower convex channel (2) is a V shape or a U shape.

3. A microfluidic chip adaptable to a fluorescent quantitative PCR instrument according to claim 2, characterized in that: The upper convex channel (3) is in an inverted V-shape, the lower convex channel (2) is in a V-shape, and the reaction pool (5) is connected to the corner of the V-shaped channel.

4. A microfluidic chip adaptable to a fluorescent quantitative PCR instrument according to claim 2, characterized in that: The upper convex channel (3) is in an inverted V-shape, the lower convex channel (2) is in a U-shape, and the reaction pool (5) is connected to the bottom of the U-shaped channel.

5. The microfluidic chip adaptable to a fluorescence quantitative PCR instrument according to claim 2, characterized in that: Any one of the lower convex channels (2) of the microfluidic channel (4) and the tops of the two adjacent upper convex channels (3) can form a liquid separation unit, each liquid separation unit is connected to one of the reaction pools (5), and the number of liquid separation units in each microfluidic channel (4) is at least 2.

6. The microfluidic chip adaptable to a fluorescence quantitative PCR instrument according to claim 2, characterized in that: The height difference between the upper convex part and the lower convex part of the bent microfluidic channel (4) is H value, and the range of H value is: 0.1mm to 5mm.

7. The microfluidic chip adaptable to a fluorescence quantitative PCR instrument according to claim 1, characterized in that: The microfluidic chip further comprises a cover sheet (8) seamlessly connected to the chip body (1), and the shape of the cover sheet (8) is adapted to the shape of the chip body (1).

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