Integrated droplet microfluidic chip, and integrated nucleic acid extraction and testing apparatus

Abstract

An integrated droplet microfluidic chip, comprising a chip body. The chip body is provided with a sample introduction channel, a continuous-phase channel, a droplet generation mechanism, a heating mechanism, a storage amplification mechanism and a tail end connection port, wherein the sample introduction channel and the continuous-phase channel are both connected to the droplet generation mechanism; the droplet generation mechanism is connected to the storage amplification mechanism; the heating mechanism is arranged in an area on the microfluidic chip corresponding to the storage amplification mechanism, and the heating mechanism comprises heating columns that are arranged in an array and all penetrate the chip body from top to bottom; and the storage amplification mechanism is connected to the tail end connection port. Further provided is an integrated nucleic acid extraction and testing apparatus. The apparatus comprises a sampling tip, a sampling gun body, a continuous-phase reservoir and the integrated droplet microfluidic chip. The problems of a sample easily being contaminated or contaminating the environment during a transfer process, a testing process being complicated and a testing platform being too large are thus solved.

Description

Integrated droplet microfluidic chip, nucleic acid extraction and detection device Technical Field

[0001] The present invention relates to the technical field of microfluidic chips, in particular to an integrated droplet microfluidic chip and a nucleic acid extraction and detection device. Background Art

[0002] Droplet microfluidics has recently gained widespread application in the field of biological testing. Nucleic acid testing, a common bioassay method, typically involves sampling a sample to generate tiny droplets. These droplets are then heated to amplify the nucleic acid. The amplified droplets are then placed on a fluorescence detection platform for observation and statistical analysis, thereby inferring the concentration of the target analyte. In short, the droplet microfluidic chip, heating platform, and fluorescence detection platform are all co-convergent and intermediate platforms for achieving biological testing.

[0003] However, the independent platforms in these methods make the sample transfer between them highly susceptible to environmental contamination. Furthermore, the entire testing process is complex and involves numerous instruments. Furthermore, there is the risk of sample leakage during transfer, which could contaminate the environment. Furthermore, the entire platform is bulky; while the droplet microfluidic chip is relatively small, its peripheral connections are complex.

[0004] Therefore, there is an urgent need for a technology that can integrate microfluidic chips and peripheral interfaces, optimize and integrate various platforms, and make such biological detection platforms more lightweight. Summary of the Invention

[0005] In view of the shortcomings of the prior art described above, the purpose of the present invention is to provide an integrated droplet microfluidic chip and nucleic acid extraction and detection device to solve the problems in the prior art that the sample transfer process is easily contaminated or pollutes the environment, the detection process is complicated, and the detection platform is bulky.

[0006] To achieve the above-mentioned objectives and other related objectives, the present invention provides an integrated droplet microfluidic chip, including a chip body, wherein the chip body is provided with an injection channel, a continuous phase channel, a droplet generating mechanism, a heating mechanism, a storage and amplification mechanism, and a tail end connection port, wherein the injection channel and the continuous phase channel are both connected to the droplet generating mechanism, the droplet generating mechanism is connected to the storage and amplification mechanism, and the heating mechanism is provided on the microfluidic chip in the corresponding area of ​​the storage and amplification mechanism, wherein the heating mechanism includes heating columns arranged in an array, and the heating columns all pass through the chip body up and down, and the storage and amplification mechanism is connected to the tail end connection port.

[0007] In an embodiment of the present invention, the injection channel is suitable for transporting samples.

[0008] In an embodiment of the present invention, the continuous phase channel is suitable for conveying the continuous phase.

[0009] In an embodiment of the present invention, the tail end connection port is suitable for connecting to a negative pressure device.

[0010] In an embodiment of the present invention, the heating column is filled with a heat-conducting material.

[0011] In an embodiment of the present invention, the integrated droplet microfluidic chip further comprises a heating plate, which is disposed at the upper and lower ends of the arrayed heating columns, and is tightly connected to the ends of each heating column.

[0012] In an embodiment of the present invention, the droplet generating mechanism is a cross-shaped flow channel, which includes a main channel and side channels on both sides of the main channel, the injection channel is connected to the main channel, and the continuous phase channel is connected to the side channels on both sides.

[0013] The present invention also provides an integrated nucleic acid extraction and detection device, comprising a sampling head, a sampling gun body, a continuous phase liquid storage tank, and the integrated droplet microfluidic chip as described above, wherein the sampling head is connected to one end of the sampling gun body, the integrated droplet microfluidic chip and the continuous phase liquid storage tank are detachably assembled in the cavity of the sampling gun body, the continuous phase channel of the integrated droplet microfluidic chip is connected to the continuous phase liquid storage tank, and the other end of the sampling gun body is suitable for connecting to a negative pressure device.

[0014] In an embodiment of the present invention, a sampling tube is provided in the sampling head, and the sampling tube is connected to the sampling channel.

[0015] In an embodiment of the present invention, the tail end connection port on the integrated droplet microfluidic chip in the gun body cavity of the sampling gun is connected to the negative pressure device.

[0016] In an embodiment of the present invention, the negative pressure device is a negative pressure pump.

[0017] As described above, the integrated droplet microfluidic chip and nucleic acid extraction and detection device of the present invention have the following beneficial effects:

[0018] The integrated droplet microfluidic chip of the present invention integrates a droplet generation mechanism, a heating mechanism, and a storage and amplification mechanism. Sheep samples are transported to the droplet generation mechanism through an injection channel, where tiny droplets are generated. Then, under the action of the heating mechanism, the tiny droplets begin to heat up rapidly and evenly in the storage and amplification mechanism for amplification, thus achieving the completion of sampling, droplet generation, and nucleic acid amplification on the same chip.

[0019] The integrated nucleic acid extraction and detection device of the present invention integrates a microfluidic chip with an external connection structure sampling head, a sampling gun body, and a continuous phase liquid storage tank, and integrates the steps of sampling, droplet generation, heating, and droplet storage into an independent component. Combined with a negative pressure device, a miniaturized detection platform is formed. After the sample is amplified in the integrated droplet microfluidic chip, the chip is directly removed for fluorescence detection and output of an analysis report, which improves the reaction and detection speed and realizes simple semi-automatic detection. It not only avoids contamination during the sample transfer process and leakage that pollutes the environment, but also the integrated nucleic acid extraction and detection device of the present invention is simple and compact.

[0020] The integrated nucleic acid extraction and detection device of the present invention can be applied to the precise quantitative detection of pathogens. The detection process only requires three simple operation steps: inserting the microfluidic chip, installing the sampling tube and the negative pressure pump, and then sampling for droplet generation and nucleic acid amplification. After waiting for the reaction to be completed, the chip is removed and fluorescence detection is performed. The design of the plug-in sampling head and microfluidic chip of the present invention greatly reduces the requirements for the operator's professional skills and auxiliary equipment during the entire use process, making the present invention extremely applicable in the field of molecular diagnosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] FIG1 is a schematic diagram showing the three-dimensional structure of the integrated droplet microfluidic chip of the present invention.

[0022] FIG2 is a schematic diagram showing the planar structure of the integrated droplet microfluidic chip of the present invention.

[0023] FIG3 is a three-dimensional schematic diagram showing the integration of the integrated droplet microfluidic chip and the heating plate of the present invention.

[0024] FIG4 is a side view showing the integrated droplet microfluidic chip combined with a heating plate according to the present invention.

[0025] FIG5 is a three-dimensional schematic diagram showing the integrated droplet microfluidic chip of the present invention being loaded into the sampling gun body.

[0026] FIG6 shows a schematic diagram I of the integrated nucleic acid extraction and detection device of the present invention.

[0027] FIG. 7 shows a schematic diagram II of the integrated nucleic acid extraction and detection device of the present invention.

[0028] FIG8 is a schematic structural diagram of a cross-shaped flow channel.

[0029] Explanation of Reference Numerals 1 Injection channel 2 Continuous phase channel 3 Droplet generation mechanism 4 Heating mechanism 5 Storage and amplification mechanism 6 Tail end connection port 7 Continuous phase reservoir 31 Main channel 32 Side channels on both sides of the left and right channels 41 Heating column 42 Heating plate 10 Sampling head 20 Sampling gun body 101 Sampling tube DETAILED DESCRIPTION

[0030] The following describes the embodiments of the present invention through specific examples. Those skilled in the art will readily understand the other advantages and benefits of the present invention from the disclosure herein. The present invention may also be implemented or applied through various other specific embodiments, and the details in this specification may be modified or altered based on different viewpoints and applications without departing from the spirit of the present invention.

[0031] In the present invention, unless otherwise clearly specified and limited, the terms "installed", "connected", "connected", "fixed" and the like should be understood in a broad sense. For example, they can refer to fixed connections, detachable connections, or integrated connections; they can refer to mechanical connections, electrical connections, or communication connections; they can refer to direct connections or indirect connections through an intermediate medium; they can refer to internal connections between two elements or interactions between two elements, unless otherwise clearly defined. At the same time, the terms "center", "longitudinal", "transverse", "upper", "lower", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside" and the like indicate directions or positional relationships based on the directions or positional relationships shown in the accompanying drawings. They are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the devices or elements referred to must have a specific direction, be constructed and operated in a specific direction. Therefore, they should not be understood as limiting the present invention. For those skilled in the art, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0032] Please refer to the accompanying drawings. It should be noted that the illustrations provided in this embodiment are merely schematic illustrations of the basic concept of the present invention. Therefore, the drawings only show components relevant to the present invention and are not drawn according to the number, shape, and size of components in actual implementation. In actual implementation, the type, quantity, and proportion of each component may be varied arbitrarily, and the component layout may also be more complex.

[0033] In a first aspect, the present invention provides an integrated droplet microfluidic chip, comprising a chip body, the chip body being provided with an injection channel 1, a continuous phase channel 2, a droplet generation mechanism 3, a heating mechanism 4, a storage and amplification mechanism 5, and a tail end connection port 6. The injection channel 1 and the continuous phase channel 2 are both connected to the droplet generation mechanism 3, which is in turn connected to the storage and amplification mechanism 5. The heating mechanism 4 is provided on the microfluidic chip in the area corresponding to the storage and amplification mechanism 5. The heating mechanism 4 comprises an array of heating posts 41, which extend vertically through the chip body. The storage and amplification mechanism 5 is connected to the tail end connection port 6. As shown in Figures 1 and 2 .

[0034] The injection channel 1 is suitable for delivering samples. The collected samples are delivered to the droplet generation mechanism through the injection channel 1.

[0035] The continuous phase channel 2 is suitable for conveying the continuous phase. The desired continuous phase liquid is conveyed to the droplet generation mechanism through the continuous phase channel 2. The tail end connection port 6 is suitable for connecting to a negative pressure device. The tail end connection port 6 at the tail of the chip is connected to the negative pressure device. Under the action of the negative pressure device, the sample entering from the injection channel 1 is uniformly fed into the storage and amplification mechanism 5 after being generated into tiny droplets by the droplet generation mechanism. Then, under the action of the heating mechanism 4, the heat from the heating column 41 causes the storage and amplification mechanism 5 to heat up rapidly and uniformly, and the nucleic acid is rapidly amplified in this area.

[0036] The storage and amplification mechanism 5 of this embodiment of the present invention comprises a flat, enclosed cavity, which is provided with a plurality of cylindrical through-holes extending through the upper and lower surfaces of the cavity. The heating rods 41 extend through these cylindrical through-holes and extend through the storage and amplification mechanism 5. The cylindrical through-holes include cylindrical walls that are sealed or integrally formed with the upper and lower surfaces of the flat, enclosed cavity to prevent leakage. Typically, the walls of the cylindrical through-holes should conform to the heating rods, allowing the heating rods 41 to fit within the cylindrical through-holes and transfer heat to the storage and amplification mechanism 5.

[0037] The heating columns 41 of the present invention may be arranged in various arrays. In one embodiment, the array is arranged in a rectangular array, or in other array forms, such as a ring array.

[0038] The heating column 41 of the present invention can have various shapes, such as cylindrical, square, curved, etc. In a preferred embodiment, its shape should match the cylindrical through hole of the storage amplification mechanism 5 .

[0039] In the integrated droplet microfluidic chip of the present invention, the heating column 41 is filled with heat-conducting material.

[0040] In a preferred embodiment of the present invention, as shown in Figures 3 and 4 , the integrated droplet microfluidic chip further includes heating plates 42, which are positioned at the upper and lower ends of the arrayed heating posts 41 and are tightly connected to the ends of each heating post 41. The heating posts 41 transfer heat to the chip, making it sensitive to temperature changes and allowing for rapid heating and cooling, while also ensuring more uniform temperature variations throughout the storage and amplification mechanism.

[0041] The heating plate 42 can be connected to a power source for heating, such as an external power source or a battery.

[0042] In the integrated droplet microfluidic chip of the present invention, as shown in Figure 8, the droplet generation mechanism 3 is a cross-shaped flow channel, which includes a main channel 31 and side channels 32 on both sides. The injection channel 1 is connected to the main channel 31, and the continuous phase channel 2 is connected to the side channels 32 on both sides. The collected sample is transported to the main channel 31 of the droplet generation mechanism through the injection channel 1, and the continuous phase is transported to the side channels 31 on both sides of the droplet generation mechanism through the continuous phase channel 2. When the sample passes through the cross-shaped droplet generation structure 3, it is cut into tiny droplets by the continuous phase in the side channels 32 on both sides. The tiny droplets then evenly enter the storage and amplification mechanism 5 area for amplification.

[0043] A second aspect of the present invention provides an integrated nucleic acid extraction and detection device, comprising a sampling head 10, a sampling gun body 20, a continuous phase reservoir 7, and the integrated droplet microfluidic chip described above. The sampling head 10 is connected to one end of the sampling gun body 20. The integrated droplet microfluidic chip and the continuous phase reservoir 7 are detachably assembled within the cavity of the sampling gun body 20. The continuous phase channel 2 of the integrated droplet microfluidic chip is connected to the continuous phase reservoir 7. The other end of the sampling gun body 20 is suitable for connection to a negative pressure device, as shown in Figures 6 and 7.

[0044] The continuous phase in the continuous phase reservoir 7 can be oil. The provision of the continuous phase reservoir 7 allows direct sampling using the sampling head 10 during actual sampling without the need to prepare another liquid. The sample obtained can be directly formed into tiny droplets under the action of the continuous phase when passing through the droplet generation mechanism 3.

[0045] In the integrated nucleic acid extraction and detection device of the present invention, the sampling head 10 is provided with a sampling tube 101 , and the sampling tube 101 is connected to the sampling channel 1 .

[0046] In the integrated nucleic acid extraction and detection device of the present invention, the tail connection port 6 on the integrated droplet microfluidic chip in the sampling gun body cavity is connected to the negative pressure device. In a preferred embodiment of the present invention, the negative pressure device is a negative pressure pump.

[0047] During use, the heating plate 42 is pressed against the ends of the heating columns 41 filled with thermally conductive material on the microfluidic chip, and the continuous phase reservoir 7 is connected to the continuous phase channel 2, as shown in Figures 3 and 4. The structure assembled as shown in Figures 3 and 4 is inserted into the cavity of the sampling gun body 20, as shown in Figure 5. The sampling tube 101 is connected to the injection channel 1, and the sampling head 10 is connected to the sampling gun body 20, as shown in Figures 6 and 7. Then, the tail end connection port 6 at the other end of the sampling gun body 20 is connected to a negative pressure pump to form a complete integrated nucleic acid extraction and detection device, and sampling can begin. A negative pressure pump is used to draw samples into the integrated droplet microfluidic chip. When the sample passes through the cross-shaped droplet generation structure, it is cut into tiny droplets by the continuous phase oil in the flow channels on both sides. The tiny droplets evenly enter the storage and amplification mechanism. At the same time, the heating plate transfers heat to the area of ​​the chip storage and amplification mechanism in a timely and uniform manner through the arrayed heating columns, and nucleic acid amplification is rapidly performed in this area. After the reaction is complete, the heating plate is turned off, the sampling tube is removed, the chip is pulled out, and the area of ​​the chip storage and amplification mechanism is placed under an optical instrument for fluorescence detection to obtain a final result report.

[0048] Example 1

[0049] An integrated droplet microfluidic chip: includes an injection channel 1, a continuous phase channel 2, a droplet generating mechanism 3, a heating mechanism 4, a storage and amplification mechanism 5 and a tail end connection port 6. The injection channel 1 and the continuous phase channel 2 are both connected to the droplet generating mechanism 3. The droplet generating mechanism 3 is a cross-shaped flow channel, which includes a main channel 31 and side flow channels 32 on both sides of the left and right channels. The injection channel 1 is connected to the main channel 31, and the continuous phase channel 2 is connected to the side flow channels 32 on the left and right sides. The droplet generating mechanism 3 is connected to the storage and amplification mechanism 5. A heating mechanism 4 is provided on the microfluidic chip corresponding to the storage and amplification mechanism 5. The heating mechanism 4 includes cylindrical heating columns 41 and a heating plate 42 arranged in a rectangular array. The heating columns 41 are filled with heat-conducting material. The heating columns 41 all pass through the microfluidic chip from top to bottom. The heating plates 42 are arranged at the upper and lower ends of the heating columns 41 arranged in the array. The heating plates 42 are tightly connected to the ends of each heating column 41. The storage and amplification mechanism 5 is connected to the tail end connection port 6. The injection channel 1 delivers the sample, the continuous phase channel 2 delivers the continuous phase, and the tail connection port 6 is connected to the negative pressure device, as shown in FIG3 .

[0050] Example 2

[0051] An integrated nucleic acid extraction and detection device includes a sampling head 10, a sampling gun body 20, a continuous phase liquid reservoir 7, and the integrated droplet microfluidic chip of Example 1. The sampling head 10 is connected to one end of the sampling gun body 20. The integrated droplet microfluidic chip and the continuous phase liquid reservoir 7 are detachably assembled in the cavity of the sampling gun body 20. A sampling tube 101 is provided in the sampling head 10, and the sampling tube 101 is connected to the injection channel 1. The continuous phase channel 2 of the integrated droplet microfluidic chip is connected to the continuous phase liquid reservoir 7. The tail connection port on the integrated droplet microfluidic chip in the cavity of the sampling gun body 20 is connected to a negative pressure pump.

[0052] In summary, the integrated droplet microfluidic chip of the present invention integrates the droplet generation mechanism, heating mechanism, and storage and amplification mechanism into one, thereby achieving the completion of sampling, droplet generation, and nucleic acid amplification on the same chip; then the integrated nucleic acid extraction and detection device prepared by combining the integrated droplet microfluidic chip with the sampling head, the sampling gun body, and the continuous phase liquid storage tank integrates the steps of sampling, droplet generation, heating, and droplet storage into an independent component, and is used in combination with a negative pressure device to form a miniaturized detection platform. After the sample is amplified in the integrated droplet microfluidic chip, the chip is directly removed for fluorescence detection and output analysis report, thereby achieving simple semi-automatic detection. Moreover, the integrated nucleic acid extraction and detection device of the present invention can adapt to situations with large detection demands and can achieve rapid distributed deployment. Therefore, the present invention effectively overcomes the various shortcomings of the prior art and has high industrial utilization value.

[0053] The above embodiments are merely illustrative of the principles and effects of the present invention and are not intended to limit the present invention. Anyone skilled in the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by one of ordinary skill in the art without departing from the spirit and technical principles disclosed herein are intended to be covered by the claims of the present invention.

Claims

1. An integrated droplet microfluidic chip, comprising a chip body, characterized in that: The chip body is provided with an injection channel (1), a continuous phase channel (2), a droplet generation mechanism (3), a heating mechanism (4), a storage amplification mechanism (5) and a tail end connection port (6); the injection channel (1) and the continuous phase channel (2) are both connected to the droplet generation mechanism (3); the droplet generation mechanism (3) is connected to the storage amplification mechanism (5); the heating mechanism (4) is provided on the microfluidic chip in the corresponding area of ​​the storage amplification mechanism (5); the heating mechanism (4) comprises heating columns (41) arranged in an array; the heating columns (41) all penetrate the chip body from top to bottom; and the storage amplification mechanism (5) is connected to the tail end connection port (6).

2. The integrated droplet microfluidic chip according to claim 1, characterized in that: The injection channel (1) is suitable for conveying samples; And / or, the continuous phase channel (2) is suitable for conveying the continuous phase.

3. The integrated droplet microfluidic chip according to claim 1, characterized in that: The tail end connection port (6) is suitable for connecting to a negative pressure device.

4. The integrated droplet microfluidic chip according to claim 1, characterized in that: The heating column (41) is filled with heat-conducting material.

5. The integrated droplet microfluidic chip according to claim 4, characterized in that: It also comprises a heating plate (42), wherein the heating plate (42) is arranged at the upper and lower ends of the heating columns (41) arranged in an array, and the heating plate (42) is tightly connected to the end of each heating column (41).

6. The integrated droplet microfluidic chip according to claim 1, characterized in that: The droplet generation mechanism (3) is a cross-shaped flow channel, which includes a main flow channel (31) and side flow channels (32) on both sides of the left and right channels. The injection channel (1) is connected to the main flow channel (31), and the continuous phase channel (2) is connected to the side flow channels (32) on the left and right sides.

7. An integrated nucleic acid extraction and detection device, characterized in that: It comprises a sampling head (10), a sampling gun body (20), a continuous phase liquid storage tank (7) and an integrated droplet microfluidic chip as described in any one of claims 1 to 6, wherein the sampling head (10) is connected to one end of the sampling gun body (20), the integrated droplet microfluidic chip and the continuous phase liquid storage tank (7) are detachably assembled in the cavity of the sampling gun body (20), the continuous phase channel (2) of the integrated droplet microfluidic chip is connected to the continuous phase liquid storage tank (7), and the other end of the sampling gun body (20) is suitable for connecting to a negative pressure device.

8. The integrated nucleic acid extraction and detection device according to claim 7, characterized in that: The sampling head (10) is provided with a sampling tube (101), and the sampling tube (101) is connected to the sampling channel (1).

9. The integrated nucleic acid extraction and detection device according to claim 7, characterized in that: The tail end connection port (6) on the integrated droplet microfluidic chip in the cavity of the sampling gun body (20) is connected to the negative pressure device.

10. The integrated nucleic acid extraction and detection device according to claim 9, characterized in that: The negative pressure device is a negative pressure pump.

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