TW202214867A - Nucleic acid detection disc and nucleic acid detection device - Google Patents

Abstract

A nucleic acid detection disc comprises a box, a detection chip, an electrophoresis box, and a barrier structure. The detection chip includes a channel, which includes a liquid outlet. The electrophoresis box is arranged in the box and the liquid outlet is used to connect the electrophoresis box with the channel. The barrier structure includes a first state and a second state. In the first state, the barrier structure is arranged on the side near the liquid outlet near the channel. The barrier structure blocks liquid outlet. In the second state, the barrier structure is far away from the liquid outlet to make the channel and the electrophoresis box connected. The nucleic acid detection box provided by the invention integrates the nucleic acid amplification reaction and electrophoresis detection, and the detection operation is simple, the efficiency is high, the detection cost is low, the detection flexibility is strong. Two kinds of liquid in the detection chip and the electrophoresis box may be avoided from accidental leakage or mixing before use. A nucleic acid detection device using the nucleic acid detection disc is also provided.

Description

Nucleic acid detection box and nucleic acid detection equipment

The present invention relates to a nucleic acid detection box and nucleic acid detection equipment.

At present, most of the detections for molecular diagnosis, morphology, immunology, etc. are carried out in professional laboratories. The traditional detection process generally includes the following steps: first perform nucleic acid amplification in large and medium-sized detection equipment; then manually transfer the amplified nucleic acid To electrophoresis detection equipment for electrophoresis detection; finally, the electrophoresis detection results are manually transferred to a special fluorescence analyzer for result analysis. The equipment required for the entire detection process is complex, bulky, low in detection efficiency, poor in flexibility, high in cost, complex in operation, and requires high professional level of operators, requiring skilled technicians to operate, which cannot achieve home portable detection. .

In view of this, in order to overcome at least one of the above-mentioned defects, it is necessary to provide a nucleic acid detection kit.

In addition, the present application also provides a nucleic acid detection device.

The present invention provides a nucleic acid detection box, the nucleic acid detection box includes a box body, a detection chip, an electrophoresis box and a blocking structure, the detection chip is arranged in the box body, and the detection chip includes a first cover plate, a spacer layer and The second cover plate, the two opposite surfaces of the spacer layer are in contact with the first cover plate and the second cover plate respectively, and the first cover plate, the spacer layer and the second cover plate are surrounded by A channel is formed, and the channel includes a liquid outlet; the electrophoresis box is arranged in the box body, and the liquid outlet is used to communicate the electrophoresis box with the channel; the blocking structure includes a first state and a In the second state, in the first state, the blocking structure is provided on the side of the liquid outlet close to the channel, and the blocking structure blocks the channel from communicating with the electrophoresis box. In the second state At the time, the blocking structure is away from the liquid outlet, so that the channel and the electrophoresis box are communicated.

In the embodiment of the present application, the electrophoresis box includes a connecting device, the channel is communicated with the electrophoresis box through the connecting device, and the connecting device includes a first end, and the first end passes through the liquid outlet A port extends into the channel, and in the first state, the blocking structure covers the first end.

In the embodiment of the present application, in the first state, the blocking structure includes a blocking part and a sealing part, the blocking part is provided in the channel, and the sealing part extends into the connection through the first end inside the device.

In the embodiment of the present application, when the temperature is 0°C to 35°C, the barrier structure is in the first state, and the first state is a solid state. When the temperature exceeds 35°C, the barrier structure is in the second state, which is a molten state.

In the embodiment of the present application, the material of the barrier structure includes a wax substance, and the wax substance includes one or more of paraffin wax, silicon wax, vegetable wax and white wax.

In the embodiment of the present application, the content of the wax substance in the barrier structure is greater than or equal to 60 wt %.

In the embodiment of the present application, the electrophoresis box further includes an electrophoresis tank, a gel medium disposed inside the electrophoresis tank, and a liquid injection tank disposed at one end of the gel medium, and the connection device includes a first The second end of one end extends into the liquid injection tank.

In the embodiment of the present application, the first end includes a flat surface or at least one inclined surface, the flat surface is parallel to the extending direction of the channel, and the inclined surface and the central axis of the connecting device are arranged obliquely.

In the embodiment of the present application, the distance between the inclined plane and the central axis of the connecting device is 45°-60°.

The present invention also provides a nucleic acid detection device, which includes a host, a detection box installation slot, and the nucleic acid detection box as described above. The detection box installation slot is arranged on the host, and the nucleic acid detection box is detachably installed with the nucleic acid detection box.

Compared with the prior art, the nucleic acid detection box provided by the present invention integrates nucleic acid amplification reaction and electrophoresis detection, the overall structure is simple, the detection operation is simple, the operation process has low professional requirements, the detection efficiency is high, and the detection cost is greatly reduced. At the same time, the detection process is flexible and does not need to be carried out in a fixed laboratory. The nucleic acid detection box is portable, which can realize community detection or home detection; the setting of the barrier structure can avoid the detection chip and electrophoresis caused by the movement or vibration of the nucleic acid detection box. The liquid in the tank is mixed or leaked, which improves the reliability of the nucleic acid detection cartridge.

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, but not all of the embodiments.

It should be noted that when an element is referred to as being "fixed to" another element, it can be directly on the other element or intervening elements may also be present. When an element is referred to as being "connected" to another element, it can be directly connected to the other element or intervening elements may also be present. When an element is referred to as being "disposed on" another element, it can be directly disposed on the other element or intervening elements may also be present. The terms "vertical," "horizontal," "left," "right," and similar expressions are used herein for illustrative purposes only.

The system implementations described below are only illustrative, and the division of the modules or circuits is only a logical function division, and other division methods may be used in actual implementation. Furthermore, it is clear that the word "comprising" does not exclude other units or steps and the singular does not exclude the plural. Multiple units or means stated in the system claim may also be implemented by the same unit or means through software or hardware. The terms first, second, etc. are used to denote names and do not denote any particular order.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terms used herein in the description of the present invention are for the purpose of describing specific embodiments only, and are not intended to limit the present invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

Please refer to FIG. 1 , FIG. 3 to FIG. 5 , and referring to FIG. 8 in conjunction with FIG. 8 , which is a nucleic acid detection cartridge 100 provided in an embodiment of the present invention, and the nucleic acid detection cartridge 100 is used for nucleic acid detection. The nucleic acid detection box 100 includes a box body 1 , a detection chip 2 , an electrophoresis box 3 and a connector 4 . The detection chip 2 is disposed in the box body 1 , and the detection chip 2 includes a first cover plate 21 , a spacer layer 22 and a second cover plate 23 . The two opposite surfaces of the spacer layer 22 are respectively connected to the first cover plate 21 and the second cover plate 23 . The second cover plate 23 is in contact, and the first cover plate 21 , the spacer layer 22 and the second cover plate 23 are surrounded to form a channel 5 , and the channel 5 is used to carry the detection liquid a. The electrophoresis box 3 is arranged in the box body 1 and communicates with the channel 5 . The connector 4 is electrically connected with the detection chip 2 and the electrophoresis box 3 respectively, and the connector 4 is used for electrical connection with the external control board. The nucleic acid detection box 100 is used for nucleic acid amplification reaction and electrophoresis detection, and the detection solution a containing nucleic acid samples is added into the channel 5 of the detection chip 2. It should be noted that the detection solution a in the channel 5 is a liquid bead If the detection solution a carries out a nucleic acid amplification reaction in the channel 5 to obtain a nucleic acid amplification product b, the nucleic acid amplification product b directly enters the electrophoresis box 3 from the detection chip 2 for electrophoresis detection, and finally uses An image of the electrophoresis box 3 is captured by the image acquisition device matched with the nucleic acid detection box 100 , wherein the image is a fluorescent photo of the electrophoresis detection. By integrating the detection chip 2 and the electrophoresis box 3 in one box body 1, the present invention has a simple overall structure, does not require complex large-scale equipment, and has low cost, and the detection solution a can directly enter the electrophoresis box 3 for electrophoresis after nucleic acid amplification is completed. It simplifies the process of sample transfer, coordination and connection in different detection links, and improves the detection efficiency.

Please refer to FIG. 1 to FIG. 5 , the box body 1 includes a first casing 11 , a second casing 12 , a sample injection port 13 disposed on the second casing 12 , and a detection device disposed on the first casing 11 . window 14. The first casing 11 and the second casing 12 together form an accommodating cavity (not shown), and the detection chip 2 , the electrophoresis box 3 and the connector 4 are all accommodated in the accommodating cavity. The sample introduction port 13 is provided corresponding to the detection chip 2 , and is used for adding the detection solution a containing the nucleic acid sample into the detection chip 2 . The detection window 14 is disposed corresponding to the electrophoresis box 3 , and the image acquisition device can collect the image of the electrophoresis box 3 through the detection window 14 .

Referring to FIG. 3 , in this embodiment, the first casing 11 and the second casing 12 are connected by means of snapping. In addition, after the first casing 11 and the second casing 12 are engaged, they can also be The periphery is fastened with screws to increase the connection firmness of the first casing 11 and the second casing 12 .

Please refer to FIG. 1 and FIG. 2. In this embodiment, the side wall of the box body 1 is further provided with an opening 17. The opening 17 is used to install the connector 4. The connector 4 is integrally located in the accommodating cavity and is connected by the opening. 17 exposes the box body 1 so as to facilitate the electrical connection between the connector 4 and the external control board.

Please refer to FIG. 2 , in this embodiment, the box body 1 further includes a card slot 15 arranged on the first housing 11 . Since the nucleic acid detection box 100 needs to be installed in the nucleic acid detection device during use, the card slot is designed 15. It is convenient to install the nucleic acid detection box 100 in the nucleic acid detection equipment used.

Please refer to FIG. 1 , in conjunction with FIG. 13 , in this embodiment, an indication mark 18 (such as an arrow) is further provided on the side of the second housing 12 away from the accommodating cavity. Referring to FIG. 13 , the indication mark 18 is used to indicate The nucleic acid detection box 100 is inserted into the insertion direction of the nucleic acid detection device 200 to avoid wrong insertion.

Please refer to FIG. 3 , in this embodiment, the box body 1 is provided with a number of supporting structures 16 . Since the detection chip 2 , the electrophoresis box 3 and the connector 4 have different thicknesses in the structural design, they are installed in the box body 1 . It is necessary to design several support structures 16 with different heights to support the detection chip 2 , the electrophoresis box 3 and the connector 4 , so as to improve the connection stability between the detection chip 2 , the electrophoresis box 3 and the connector 4 .

In this embodiment, the box body 1 is made of plastic material, wherein the support structure 16 is integrally formed with the first casing 11 and the second casing 12 .

Please refer to FIG. 4 and FIG. 5 , the detection chip 2 further includes a drive circuit 24 disposed on the side of the first cover plate 21 close to the second cover plate 23 , and a drive circuit 24 disposed on the side of the drive circuit 24 close to the second cover plate 23 . The first dielectric layer 26 on the side, the conductive layer 25 disposed on the side of the second cover plate 23 close to the first cover plate 21 , and the second dielectric layer 25 disposed on the side of the conductive layer 25 close to the first cover plate 21 The electrical layer 27, the driving circuit 24 and the conductive layer 25 are all electrically connected to the connector 4, and the detection liquid a can be realized in the channel 5 by energizing or de-energizing the driving circuit 24 and the conductive layer 25 Move along the prescribed path.

Please refer to FIG. 5 and FIG. 6 , in conjunction with FIG. 1 , in this embodiment, the driving circuit 24 includes a plurality of driving electrodes 241 arranged in an array and a control electrode 242 electrically connected to all the driving electrodes 241 . 242 is electrically connected to the connector 4 . Specifically, the driving circuit 24 is a thin film transistor (Thin Film Transistor, TFT) driving circuit, and since the detection liquid a has conductivity, combined with the principle of dielectric wetting (Electrowitting-On-Dielectric, EWOD), the detection liquid can be realized. a Move according to the specified path in the channel 5. Using the TFT principle, the circuit between a certain driving electrode 241 and the conductive layer 25 can be selectively turned on or off, thereby changing the voltage between the driving electrode 241 and the conductive layer 25 to change the detection solution a and the first dielectric layer. 26 and the second dielectric layer 27 between the wetting characteristics, and then control the detection liquid a in the channel 5 to move according to a predetermined path. As shown in FIG. 5 , the detection solution a moves on the electrode I, the electrode H and the electrode G. When the detection solution a is on the electrode H, a voltage is applied between the electrode G and the conductive layer 25 to give the electrode G a voltage Vd, and at the same time The voltage between the electrode H and the conductive layer 25 is disconnected, and the wetting characteristics between the detection liquid a and the first dielectric layer 26 and the second dielectric layer 27 are changed, so that the electrode H and the detection liquid a are in contact with each other. The liquid-solid contact angle between the electrodes becomes larger, and the liquid-solid contact angle between the electrode G and the detection liquid a becomes smaller, so that the detection liquid a moves from the electrode H to the electrode G.

In the present embodiment, the first dielectric layer 26 and the second dielectric layer 27 are both insulating and hydrophobic layers, specifically, a polytetrafluoroethylene coating. It can make the detection liquid a move more smoothly in the predetermined path, and avoid the liquid bead breaking during the moving process.

In this embodiment, referring to FIG. 5 , the driving circuit 24 is disposed on the side of the first cover plate 21 close to the channel 5 . Specifically, the driving circuit 24 may be formed by a metal etching method or an electroplating method.

In this embodiment, the control electrode 242 is integrated on the same edge of the first cover plate 21 , and the detection chip 2 and the detection chip 2 are realized by inserting the side of the first cover plate 21 on which the control electrode 242 is disposed into the connector 4 . Electrical connection of the connector 4 .

Please refer to FIG. 6 , in conjunction with FIG. 3 and FIG. 8 , the driving circuit 24 can be divided into a plurality of areas according to different purposes, namely a sample adding area A, a reagent storage area B, a plurality of nucleic acid amplification areas C and a liquid discharge area. D. The detection chip 2 is also provided with a chip sample adding groove 6 corresponding to the sample adding area A, the chip sample adding groove 6 is communicated with the sample adding area A, and at the same time, the chip sample adding groove 6 is connected with the sample adding groove on the second cover 23 . Corresponding to the sample port 13 , the detection solution a is added to the sample adding area A from the sample adding port 13 . The reagent storage area B is used to store fluorescent reagents (such as fluorescent dyes or fluorescent probes). The detection solution a performs a nucleic acid amplification reaction in the nucleic acid amplification region C. The nucleic acid amplification region C may include multiple regions, and the number of specific regions may be determined according to actual detection requirements. The liquid outlet area D includes a liquid outlet 51 , the channel 5 communicates with the electrophoresis box 3 through the liquid outlet 51 , and the nucleic acid amplification product b can enter the electrophoresis box 3 in the liquid outlet area D through the liquid outlet 51 . Electrophoresis detection was performed inside.

Please refer to FIG. 5 and FIG. 6 , in conjunction with FIG. 8 , the specific movement path of the detection liquid a in the detection chip 2 is: after the detection liquid a enters the sample adding area A, it moves to the nucleic acid amplification according to a specified path under the driving of the driving electrode 241 The amplification reaction is carried out in the expansion area C; when the amplification reaction is completed, the amplified product moves to the reagent storage area B to be mixed with the fluorescent reagent, thereby obtaining the nucleic acid amplification product b combined with the fluorescent reagent; The increased product b moves to the liquid outlet area D under the driving of the driving electrode 241 , and enters the electrophoresis box 3 through the liquid outlet 51 of the liquid outlet area D.

In this embodiment, the number of the nucleic acid amplification regions C is two, and the heating temperatures of the two nucleic acid amplification regions C are different, so that different stages of the nucleic acid amplification reaction of the detection solution a at different temperatures can be realized.

In this embodiment, the fluorescent reagent is pre-coated in the reagent storage area B when the detection chip 2 is assembled, and the fluorescent reagent does not need to be added separately afterwards.

Please refer to FIG. 3 , in other embodiments, the fluorescent reagent can also be mixed with the amplification product by subsequent addition. Specifically, a reagent tank 7 is provided on the detection chip 2 corresponding to the reagent storage area B, and a fluorescent reagent can be added to the reagent tank 7 during nucleic acid detection. This design can select the type of fluorescent reagent according to actual needs. , which improves the flexibility of nucleic acid amplification reactions.

Please refer to FIG. 3 , FIG. 5 and FIG. 6 , the detection chip 2 further includes a heating element 28 disposed on the side of the first cover plate 21 and/or the second cover plate 23 away from the channel 5 . The nucleic acid amplification area C should be set for heating the detection solution a. The heating element 28 is electrically connected to the connector 4 , and a specific area of the channel 5 is heated by the heating element 28 .

In this embodiment, the heating element 28 is disposed on the side of the first cover plate 21 and the second cover plate 23 away from the channel 5 .

In this embodiment, the heating element 28 is bonded to the surfaces of the first cover plate 21 and the second cover plate 23 by thermally conductive adhesive.

Please refer to FIG. 3 and FIG. 5 , in this embodiment, the first cover plate 21 and the second cover plate 23 are both glass plates, and the spacer layer 22 is a double-sided tape frame, which is pasted on the first cover by the double-sided tape frame. The edges of a cover plate 21 and a second cover plate 23 together form a sealed channel 5 . The capacity of the channel 5 can be adjusted by designing spacer layers 22 with different thicknesses according to actual needs.

In this embodiment, after the detection chip 2 is assembled, silicone oil (not shown) will be injected into the channel 5, and the detection liquid a will move in the silicone oil according to a predetermined path.

Please refer to FIG. 3 , FIG. 4 , FIG. 7 and FIG. 8 , the electrophoresis box 3 includes an electrophoresis tank 31 , electrophoresis electrodes 32 disposed at both ends of the electrophoresis tank 31 , a gel medium 33 disposed inside the electrophoresis tank 31 , and a gel The liquid injection tank 34 at one end of the medium 33 , the connection device 35 , and the wetting liquid set in the electrophoresis tank 31 . The electrophoresis electrode 32 is electrically connected to the connector 4 , the connection device 35 includes a first end 351 and a second end 352 , the first end 351 extends into the channel 5 through the liquid outlet 51 , and the second end 352 Extending into the liquid injection tank 34 , the nucleic acid amplification product b will enter the connecting device 35 through the liquid outlet 51 in the liquid outlet area D. Further, it enters into the liquid injection tank 34 of the gel medium 33 to perform electrophoresis detection.

Please refer to FIG. 3 , FIG. 4 , FIG. 7 and FIG. 8 , in this embodiment, the electrophoresis tank 31 is located on the side of the first cover plate 21 away from the second cover plate 23 , and the opening of the electrophoresis tank 31 faces the One side of the first cover plate 21 , and the opening of the electrophoresis tank 31 faces the first cover plate 21 . The electrophoresis tank 31 includes a transparent bottom plate 311 and a plurality of side walls 312 connected to the transparent bottom plate 311 . The first cover plate 21 serves as a cover plate of the electrophoresis tank 31 to seal the electrophoresis box 3 . The above design makes a height difference ΔH ₁ between the silicone oil in the detection chip 2 and the wetting solution in the electrophoresis box 3, so that the nucleic acid amplification product b in the channel 5 can smoothly enter the electrophoresis box 3; in addition, this structure The design can improve space utilization, which is beneficial to reduce the volume of the overall nucleic acid detection box 100 .

In this embodiment, a sealing rubber ring (not shown) is disposed between the side wall 312 and the first cover plate 21 to improve the sealing performance of the electrophoresis box 3 .

Please refer to FIG. 7 , the electrophoresis tank 31 further includes a plurality of latching positions 313 disposed on the transparent bottom plate 311 , and the gel medium 33 is substantially a rectangular parallelepiped structure, which can be clamped between the plurality of the latching positions 313 . The design of the clamping position 313 can prevent the gel medium 33 from moving and dislocating, thereby ensuring the accuracy of electrophoresis detection.

Referring to FIG. 3 , in this embodiment, the transparent bottom plate 311 is a transparent glass plate, and electrophoresis results can be observed.

In the present embodiment, the number of the latching elements 313 is four, and the four latching elements 313 are respectively located at four corners of the gel medium 33 of the rectangular parallelepiped structure, so as to fix the gel medium 33 .

Please refer to FIG. 4 again, the electrophoresis tank 31 further includes a liquid injection hole 36 . The liquid injection hole 36 is disposed at the position of the first cover plate 21 corresponding to the electrophoresis tank 3 , and the liquid injection hole 36 can be used for the electrophoresis tank. Inject a wetting fluid (such as buffer) into 31.

Please refer to FIGS. 8 to 10 , and in conjunction with FIG. 4 , the first end 351 of the connecting device 35 extends into the channel 5 through the liquid outlet 51 , and the liquid outlet 51 penetrates the first cover plate 21 . The connecting device 35 is a capillary tube, and the nucleic acid amplification product b in the channel 5 can enter the gel medium 33 of the electrophoresis box 3 by using the capillary effect. As shown in FIG. 8 , in order to smoothly enter the nucleic acid amplification product b into the electrophoresis box 3 , the end surface of the first end 351 needs to be flush with the liquid level of the silicone oil, that is, the first end 351 includes a flat surface. Alternatively, as shown in FIG. 9 and FIG. 10 , the first end 351 is provided with at least one inclined plane, that is, the first end 351 is disposed inclined corresponding to the central axis c of the connecting device 35 , and the lowest point of the inclined plane is at the same time as the central axis c of the connecting device 35 . There is a difference ΔH ₂ between the lower surfaces of the channels 5 , and the liquid level of the silicone oil is located on the inclined surface, which also enables the nucleic acid amplification product b to enter the connecting device 35 smoothly. During the assembly design process of the connecting device 35 and the detection chip 2, the connecting device 35 needs to be filled with a wetting solution, and the wetting solution should be able to contact the surface of the liquid bead of the nucleic acid amplification product b at the liquid outlet area D to form The continuous liquid flow can ensure that the nucleic acid amplification product b can smoothly enter the connecting device 35 by using the capillary principle.

In this embodiment, the angle between the inclined plane and the central axis c of the connecting device 35 is 45°-60°. It has been verified by experiments that within this angle range, the nucleic acid amplification product b can smoothly enter the connecting device 35 and then enter inside the gel medium 33 .

In this embodiment, as shown in FIG. 9 , the first end 351 of the connecting device 35 is formed with an inclined plane with an inclination angle α of 45°-60°. Through the design of the inclined plane and the capillary principle, the nucleic acid can be The amplification product b enters into the connecting device 35 and into the gel medium 33 smoothly.

In another embodiment, as shown in FIG. 10 , two opposite sides of the first end 351 of the connecting device 35 are respectively formed with inclined planes with an inclination angle α of 45°-60°. According to the capillary principle, the nucleic acid amplification product b can smoothly enter into the connecting device 35 and into the gel medium 33 .

Referring to FIG. 4 , one end of the electrophoresis electrode 32 extends into the electrophoresis tank 31 , and the other end is electrically connected to the connector 4 .

Please refer to FIG. 3 , and referring to FIG. 8 , since there is a height difference ΔH ₁ between the detection chip 2 and the electrophoresis box 3 , under normal and stable conditions, the wetting solution in the electrophoresis box 3 will not pass through the connecting device 35 Enter channel 5. Since the surface of the first end 351 of the connecting device 35 is just flush with the liquid level of the silicone oil, the silicone oil in the channel 5 will not enter the electrophoresis box 3 through the connecting device 35 even under stable conditions. However, when the nucleic acid detection box 100 tilts, vibrates or changes in internal pressure during transportation, it can be compared by high-altitude low-pressure (0.2-0.7 bar) and vibration tests. If there is pressure difference and vibration outside 5, the silicone oil in channel 5 and the wetting solution in electrophoresis box 3 will leak or the two will be mixed, which will seriously affect the performance of the nucleic acid detection box 100, and may even lead to nucleic acid detection. Box 100 is directly scrapped.

Please refer to FIG. 11 , in conjunction with FIG. 1 , in order to avoid accidental leakage of the silicone oil in the channel 5 and the wetting liquid in the electrophoresis box 3 or accidental mixing of the two under the above-mentioned special circumstances, the present invention solves the above problem by adding a blocking structure 8 question. The blocking structure 8 includes a first state and a second state. In the first state, the blocking structure 8 is located on the side of the liquid outlet 51 close to the channel 5 , and the blocking structure 8 can block the channel 5 of the detection chip 2 from the channel 5 . The electrophoresis box 3 is connected. In the second state, the blocking structure 8 is far away from the liquid outlet 51 , so that the channel 5 and the electrophoresis box 3 communicate with each other. By providing the blocking structure 8 which can change state, it is ensured that the silicone oil in the channel 5 of the detection chip 2 and the wetting liquid in the electrophoresis box 3 will not leak or mix before use.

The barrier structure 8 can change its state according to different external conditions such as temperature, pressure or solvent. In this embodiment, the state of the barrier structure 8 can be changed according to the change of temperature.

In this embodiment, when the temperature is 0°C to 35°C, the barrier structure 8 is in a first state, wherein the first state is a solid state. When the temperature exceeds 35°C, the barrier structure 8 is in a second state, wherein the second state is a molten state.

In this embodiment, the blocking structure 8 needs to seal the first end 351 of the connecting device 35 .

Referring to FIG. 12 , in another embodiment, in order to better achieve a sealing effect, in the first state, the blocking structure 8 may include a blocking portion 81 and a sealing portion 82 . The blocking portion 81 is disposed in the channel 5 to seal the liquid outlet 51 and the opening of the first end 351 ; the sealing portion 82 extends into the connecting device 35 through the first end 351 to further enhance the sealing effect. Preferably, the sealing portion 82 can extend to the surface of the first cover plate 21 to form a substantially T-shaped structure. This structure makes the adhesion area of the blocking structure 8 in the channel 5 and the connecting device 35 larger, and will not be affected by External force causes the blocking structure 8 to fall off unexpectedly, resulting in seal failure.

In this embodiment, the barrier structure 8 can use one or more wax-like substances with low temperature formability, such as paraffin wax, silicon wax, vegetable wax, and white wax, as the main agent, and add solvents with different fluidity to form a high barrier effect. Molecular sealing material. During the assembly process of the nucleic acid detection box 100 , the above-mentioned polymer sealing material is coated or dot-coated on the side of the liquid outlet 51 close to the channel 5 to form the above-mentioned blocking structure 8 to achieve sealing. Among them, the molecular formula of paraffin is C _n H _{₂ (n+2)} , where n=20~40, which is obtained by cold pressing or solvent dewaxing, sweating and other methods of waxy fractions of natural or artificial petroleum. Silicone wax is a waxy material with organosiloxane (Si-O-Si) functional group and other organic materials grafted and modified. Generally, the hardness is moderate, and it is lipophilic, hydrophobic, smooth, soft and shiny. effect. Vegetable waxes usually refer to oily substances with fatty acids, monovalent or divalent fatty alcohols and higher melting points, such as wood wax, soybean wax, palm wax, rice bran wax, etc. White wax oil is usually mineral oil, liquid paraffin and the like. The solvent can be a common solvent that can be matched with the main agent.

In this embodiment, the composition ratio of the polymer sealing material is roughly 0-40 wt % of the solvent, and 60-100 wt % of the main agent, which can be adjusted according to actual needs.

In this embodiment, the polymer sealing material includes 80wt% to 99wt% of paraffin or silicon wax and 1wt% to 2wt% of mineral oil or lubricating oil. The barrier structure 8 made of such a polymer sealing material can ensure that the nucleic acid detection box 100 does not dissolve, fall or deform during transportation, etc. The silicone oil and the wetting fluid in the electrophoresis box 3 will not accidentally leak or mix.

In this embodiment, the storage environment temperature of the polymer sealing material is generally 0°C to 35°C. When the nucleic acid detection cartridge 100 is not in use, the barrier structure 8 is in the first state (ie, solid state). The melting temperature of the polymer sealing material is generally 35°C to 60°C or can be adjusted up and down depending on the performance requirements. Therefore, it can be dissolved after the nucleic acid amplification reaction starts, so that the barrier structure 8 is in the second state (that is, melted). state), so that the detection chip 2 communicates with the electrophoresis box 3.

The polymer sealing material used in the blocking structure 8 of the present invention can achieve the purpose of blocking without affecting the normal operation of the nucleic acid detection box 100 . The barrier structure 8 is in a liquid state after being melted. Due to the difference in specific gravity, the barrier structure 8 floats above the silicone oil or settles under the silicone oil, and its composition is basically a passive substance, which will not react with the silicone oil. Even if it is mixed with the silicone oil, since the silicone oil and the above-mentioned polymer sealing material have the same C-H or C-H-O-Si-O structure, the biological reaction of the nucleic acid detection box 100 will not be affected. As for the wetting liquid in the electrophoresis box 3 , it is mainly a water-based buffer liquid, and oil and water are immiscible, so it will not affect the electrophoresis detection process in the electrophoresis box 3 .

The above-mentioned method of blocking the detection chip 2 and the electrophoresis box 3 of the present invention is not limited to the application scenario of the nucleic acid detection box 100 of the present application, and can also be used in other related biomedical analyzers or sensors to isolate two liquids, Avoid accidental leakage or mixing of two liquids.

Please refer to FIG. 13, the present invention also provides a nucleic acid detection device 200, the nucleic acid detection device 200 includes a host 201 and the nucleic acid detection box 100 as described above, the host 201 is provided with a detection box installation slot 202, the nucleic acid detection box 100 is installed in the detection box installation slot 202 .

Compared with the prior art, the nucleic acid detection box provided by the present invention integrates nucleic acid amplification reaction and electrophoresis detection, the overall structure is simple, the detection operation is simple, the operation process has low professional requirements, the detection efficiency is high, and the detection cost is greatly reduced. At the same time, the detection process is flexible and does not need to be carried out in a fixed laboratory. The nucleic acid detection box is portable, which can realize community detection or home detection; the setting of the barrier structure can avoid the detection chip and electrophoresis caused by the movement or vibration of the nucleic acid detection box. The liquid in the tank is mixed or leaked, which improves the reliability of the nucleic acid detection cartridge.

100: Nucleic acid detection box 1: Box body 11: First shell 12: Second shell 13: Sample injection port 14: Detection window 15: Card slot 16: Support structure 17: Opening 18: Indicator mark 2: Detection chip 21 : first cover plate 22: spacer layer 23: second cover plate 24: drive circuit 241: drive electrode 242: control electrode 25: conductive layer 26: first dielectric layer 27: second dielectric layer 28: heating element 5 : Channel 51: Liquid outlet 3: Electrophoresis box 31: Electrophoresis tank 311: Transparent bottom plate 312: Side wall 313: Clip 32: Electrophoresis electrode 33: Gel medium 34: Liquid injection tank 35: Connection device 351: First end 352 : Second end 36: Liquid injection hole 4: Connector 6: Chip loading tank 7: Reagent tank 8: Barrier structure 81: Barrier part 82: Sealing part a: Detection solution b: Nucleic acid amplification product c: Central axis A : Sample adding area B: Reagent storage area C: Nucleic acid amplification area D: Liquid discharge area 200: Nucleic acid detection equipment 201: Host 202: Detection box installation groove ΔH ₁ : Height difference ΔH ₂ : Step difference

FIG. 1 is a schematic structural diagram of a nucleic acid detection cassette provided by an embodiment of the present invention.

FIG. 2 is a schematic structural diagram of the nucleic acid detection cassette provided by an embodiment of the present invention from another angle.

FIG. 3 is an exploded view of a nucleic acid detection cartridge provided by an embodiment of the present invention.

FIG. 4 is an exploded view of the nucleic acid detection cassette provided by an embodiment of the present invention with the cassette body removed.

FIG. 5 is a schematic cross-sectional view of a detection chip according to an embodiment of the present invention.

FIG. 6 is a schematic structural diagram of a TFT driving circuit in a detection chip according to an embodiment of the present invention.

FIG. 7 is a schematic structural diagram of an electrophoresis box according to an embodiment of the present invention.

FIG. 8 is a schematic diagram of the nucleic acid amplification product entering the electrophoresis box from the connecting device according to an embodiment of the present invention.

FIG. 9 is a schematic diagram of the nucleic acid amplification product entering the electrophoresis box from the connecting device in another embodiment of the present invention.

FIG. 10 is a schematic diagram of the nucleic acid amplification product entering the electrophoresis box from the connecting device in another embodiment of the present invention.

FIG. 11 is a schematic structural diagram of a liquid outlet provided with a blocking structure according to an embodiment of the present invention.

12 is a schematic structural diagram of a barrier structure provided at a liquid outlet according to another embodiment of the present invention.

FIG. 13 is a schematic structural diagram of a nucleic acid detection device provided by an embodiment of the present invention.

100: Nucleic acid detection kit

1: Box body

11: The first shell

12: Second shell

13: Injection port

17: Opening

18: Indication sign

4: Connector

Claims (10)

A nucleic acid detection box, comprising: box body; The detection chip is arranged in the box, the detection chip includes a first cover plate, a spacer layer and a second cover plate, and the two opposite surfaces of the spacer layer are respectively connected with the first cover plate and the second cover The plates are adjacent to each other, the first cover plate, the spacer layer and the second cover plate are surrounded to form a channel, and the channel includes a liquid outlet; an electrophoresis box, arranged in the box body, and the liquid outlet is used to communicate the electrophoresis box with the channel; and The blocking structure includes a first state and a second state. In the first state, the blocking structure is arranged on the side of the liquid outlet close to the channel, and the blocking structure blocks the channel and the electrophoresis In the second state, the blocking structure is away from the liquid outlet, so that the channel and the electrophoresis box communicate with each other. The nucleic acid detection box according to claim 1, wherein the electrophoresis box includes a connecting device, the channel is communicated with the electrophoresis box through the connecting device, the connecting device includes a first end, and the first One end extends into the channel through the liquid outlet, and in the first state, the blocking structure covers the first end. The nucleic acid detection cartridge according to claim 2, wherein, in the first state, the blocking structure includes a blocking part and a sealing part, the blocking part is provided in the channel, and the sealing part passes through the first state. One end extends into the connecting device. The nucleic acid detection box according to claim 1, wherein when the temperature is 0°C to 35°C, the barrier structure is in the first state, and the first state is solid; When the temperature exceeds 35°C, the barrier structure is in the second state, which is a molten state. The nucleic acid detection kit according to claim 4, wherein the material of the blocking structure comprises a wax-like substance, and the wax-like substance comprises one or more of paraffin wax, silicon wax, vegetable wax and white wax. The nucleic acid detection kit according to claim 5, wherein the content of the wax-like substance in the barrier structure is greater than or equal to 60 wt%. The nucleic acid detection box according to claim 2, wherein the electrophoresis box further comprises an electrophoresis tank, a gel medium disposed inside the electrophoresis tank, and a liquid injection tank disposed at one end of the gel medium, the The connecting device includes a second end remote from the first end, the second end extending into the liquid injection tank. The nucleic acid detection cartridge according to claim 2, wherein the first end includes a flat surface or at least one inclined surface, the flat surface is parallel to the extending direction of the channel, and the inclined surface is located between the center axis of the connecting device Tilt setting. The nucleic acid detection cartridge according to claim 8, wherein the angle between the inclined plane and the central axis of the connecting device is 45°-60°. A nucleic acid detection device, comprising: host; a detection box installation slot, disposed on the host; and The nucleic acid detection cartridge according to any one of claims 1 to 9, wherein the nucleic acid detection cartridge is detachably installed in the detection cartridge installation slot.

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