CN114317225A - Nucleic acid detecting cassette and nucleic acid detecting apparatus - Google Patents
Nucleic acid detecting cassette and nucleic acid detecting apparatus Download PDFInfo
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- CN114317225A CN114317225A CN202110746172.3A CN202110746172A CN114317225A CN 114317225 A CN114317225 A CN 114317225A CN 202110746172 A CN202110746172 A CN 202110746172A CN 114317225 A CN114317225 A CN 114317225A
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- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 109
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 109
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 109
- 238000001962 electrophoresis Methods 0.000 claims abstract description 118
- 238000001514 detection method Methods 0.000 claims abstract description 116
- 239000007788 liquid Substances 0.000 claims abstract description 87
- 230000003321 amplification Effects 0.000 claims abstract description 44
- 238000003199 nucleic acid amplification method Methods 0.000 claims abstract description 44
- 238000006243 chemical reaction Methods 0.000 claims abstract description 11
- 125000006850 spacer group Chemical group 0.000 claims description 11
- 238000004891 communication Methods 0.000 claims description 8
- 238000002347 injection Methods 0.000 claims description 7
- 239000007924 injection Substances 0.000 claims description 7
- 238000009736 wetting Methods 0.000 description 16
- 229920002545 silicone oil Polymers 0.000 description 15
- 239000003153 chemical reaction reagent Substances 0.000 description 14
- 239000000523 sample Substances 0.000 description 14
- 238000000034 method Methods 0.000 description 11
- 239000012530 fluid Substances 0.000 description 8
- 238000010438 heat treatment Methods 0.000 description 8
- 239000000243 solution Substances 0.000 description 6
- 238000013461 design Methods 0.000 description 5
- 238000010586 diagram Methods 0.000 description 5
- 239000000853 adhesive Substances 0.000 description 3
- 230000001070 adhesive effect Effects 0.000 description 3
- 239000011324 bead Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 239000007850 fluorescent dye Substances 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- 238000003780 insertion Methods 0.000 description 2
- 230000037431 insertion Effects 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000012938 design process Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000009713 electroplating Methods 0.000 description 1
- 238000005530 etching Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000009434 installation Methods 0.000 description 1
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- 239000000203 mixture Substances 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- -1 polytetrafluoroethylene Polymers 0.000 description 1
- 229920001343 polytetrafluoroethylene Polymers 0.000 description 1
- 239000004810 polytetrafluoroethylene Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000010409 thin film Substances 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
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Abstract
A nucleic acid detection box and nucleic acid detection equipment, the nucleic acid detection box includes detecting chip, electrophoresis box and communicating structure, the detecting chip includes the channel, liquid outlet and first opening are communicated with channel; the electrophoresis box is arranged below the detection chip and comprises a liquid inlet and a second opening, the liquid inlet is arranged corresponding to the channel, the second opening is arranged far away from the channel, and the liquid outlet is communicated with the liquid inlet so that a nucleic acid amplification product enters the electrophoresis box through the channel; the two ends of the communicating structure are respectively communicated with the first opening and the second opening so as to communicate the channel with the electrophoresis box. The nucleic acid detection box provided by the invention integrates nucleic acid amplification reaction and electrophoresis detection, has the advantages of simple and convenient detection operation, high efficiency, low detection cost and strong detection flexibility, and can avoid accidental leakage or mixing of the detection chip and liquid in the electrophoresis box.
Description
Technical Field
The present invention relates to a nucleic acid detecting cassette and a nucleic acid detecting apparatus.
Background
Currently, most of the tests for molecular diagnosis, morphology, immunology, etc. are performed in professional laboratories, and the conventional testing process generally includes the following steps: firstly, carrying out nucleic acid amplification by large and medium-sized detection equipment; then, manually transferring the amplified nucleic acid to electrophoresis detection equipment for electrophoresis detection; and finally, manually transferring the electrophoresis detection result to a special fluorescence analyzer for result analysis. The whole detection process requires complicated equipment, large volume, low detection efficiency, poor flexibility, high cost, complex operation and higher requirement on the professional level of operators, and the detection process needs to be operated by skilled technicians and cannot realize household portable detection.
Disclosure of Invention
In view of the above, in order to overcome at least one of the above-mentioned drawbacks, it is necessary to provide a nucleic acid detecting cassette.
In addition, the application also provides a nucleic acid detection device.
The invention provides a nucleic acid detection box, which comprises a detection chip, an electrophoresis box and a communication structure, wherein the detection chip comprises a first cover plate, a spacing layer and a second cover plate, two opposite surfaces of the spacing layer are respectively adjacent to the first cover plate and the second cover plate, a channel is formed by the first cover plate, the spacing layer and the second cover plate in a surrounding way, the first cover plate is provided with a liquid outlet, the second cover plate is provided with a first opening, the liquid outlet and the first opening are both communicated with the channel, and the channel is used for bearing detection liquid so that the detection liquid carries out nucleic acid amplification reaction in the channel to obtain a nucleic acid amplification product. The electrophoresis box is arranged on one side of the first cover plate far away from the channel, the electrophoresis box comprises a liquid inlet and a second opening, the liquid inlet corresponds to the channel, the second opening is far away from the channel, and the liquid outlet is communicated with the liquid inlet so that the nucleic acid amplification product enters the electrophoresis box. The communicating structure is characterized in that two ends of the communicating structure are respectively communicated with the first opening and the second opening so as to communicate the channel with the electrophoresis cassette.
In an embodiment of the present application, the communicating structure includes a first connecting end communicated with the first opening, a second connecting end communicated with the second opening, and a connecting cavity connecting the first connecting end and the second connecting end.
In the embodiment of the application, the communicating structure further comprises a buffer cavity, and the buffer cavity is arranged between the second connecting end and the connecting cavity.
In the embodiment of the present application, the communicating structure is a communicating pipe.
In the embodiment of the present application, the communicating structure includes a first sidewall, a second sidewall, a third sidewall, a bottom plate and a top plate, the first sidewall is disposed on the surface of the second cover plate, the second sidewall and the third sidewall are both disposed on the surface of the electrophoresis cassette close to one side of the detection chip, the third sidewall is close to the spacer layer, the second sidewall is far away from the spacer layer, the bottom plate is disposed on the surface of the second cover plate and connected to the third sidewall, the top plate is disposed on the first sidewall and the second sidewall, the first sidewall, the second sidewall, the third sidewall, the bottom plate, the top plate and the surface of the electrophoresis cassette close to one side of the detection chip form a cavity together, the first opening penetrates through the bottom plate and is located in the cavity, the second opening is located in the cavity, and the cavity is communicated with the channel and the electrophoresis box through the first opening and the second opening respectively.
In the embodiment of the present application, the first opening is further disposed through the top plate.
In the embodiment of this application, the electrophoresis box includes a connecting device, connecting device's both ends are connected respectively the liquid outlet with the inlet, so that the passageway with electrophoresis box intercommunication, connecting device includes first end, first end via the liquid outlet stretches into the passageway.
In the embodiment of the application, the electrophoresis box further comprises an electrophoresis tank, a gel medium arranged in the electrophoresis tank and a liquid injection tank arranged at one end of the gel medium, the connecting device comprises a second end far away from the first end, and the second end extends into the liquid injection tank.
In the embodiment of the application, the first cover plate is arranged at the opening of the electrophoresis tank.
The invention also provides nucleic acid detection equipment which comprises a host machine, a detection box mounting groove and the nucleic acid detection box. The detection box mounting groove is arranged on the host machine, and the nucleic acid detection box is detachably mounted in the nucleic acid detection box mounting groove.
Compared with the prior art, the nucleic acid detection kit provided by the invention integrates nucleic acid amplification reaction and electrophoresis detection, has a simple overall structure, is simple and convenient to detect and operate, has low professional requirements on the operation process, is high in detection efficiency, and greatly reduces the detection cost; meanwhile, the detection process has strong flexibility, does not need to be carried out in a fixed laboratory, and the nucleic acid detection box is portable and can realize community detection or family detection; the arrangement of the communicating structure can avoid the liquid mixing or leakage in the detection chip and the electrophoresis tank caused by the movement or vibration of the nucleic acid detection box, and improve the reliability of the nucleic acid detection box.
Drawings
FIG. 1 is a schematic view of a nucleic acid detecting cassette according to an embodiment of the present invention.
FIG. 2 is a schematic view of another embodiment of the nucleic acid detecting cassette according to the present invention.
FIG. 3 is an exploded view of a nucleic acid detecting cassette according to an embodiment of the present invention.
FIG. 4 is an exploded view of the nucleic acid detecting cassette according to the embodiment of the present invention with the cassette removed.
Fig. 5 is a schematic cross-sectional view of a detection chip according to an embodiment of the 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 cassette according to an embodiment of the present invention.
FIG. 8 is a schematic diagram of nucleic acid amplification products entering an electrophoresis cassette from a ligation device according to an embodiment of the present invention.
FIG. 9 is a schematic diagram of nucleic acid amplification products entering an electrophoresis cassette from a ligation device according to another embodiment of the present invention.
FIG. 10 is a schematic view of nucleic acid amplification products entering an electrophoresis cassette from a ligation device according to still another embodiment of the present invention.
Fig. 11 is a sectional structure view of a communication structure provided between a first opening and a second opening according to an embodiment of the present invention.
Fig. 12 is a schematic structural diagram of a communication structure provided between a first opening and a second opening according to an embodiment of the present invention.
Fig. 13 is a schematic structural view of a communication structure provided between a first opening and a second opening according to another embodiment of the present invention.
FIG. 14 is a schematic structural view of a nucleic acid detecting apparatus according to an embodiment of the present invention.
Description of the main elements
The following detailed description will further illustrate the invention in conjunction with the above-described figures.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments.
It will be understood that when an element is referred to as being "secured to" another element, it can be directly on the other element or intervening elements may also be present. When a component is referred to as being "connected" to another component, it can be directly connected to the other component or intervening components may also be present. When a component is referred to as being "disposed on" another component, it can be directly on the other component or intervening components may also be present. The terms "vertical," "horizontal," "left," "right," and the like as used herein are for illustrative purposes only.
The system embodiments described below are merely illustrative, and the division of the modules or circuits is merely a logical division, and other divisions may be realized in practice. Furthermore, it is obvious that the word "comprising" does not exclude other elements or steps, and the singular does not exclude the plural. Several units or means recited in the system claims may also be implemented by one and the same unit or means in software or hardware. The terms first, second, etc. are used to denote names, but not any particular order.
Unless defined otherwise, 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 terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
Referring to FIG. 1, FIG. 3 to FIG. 5, and FIG. 8, a nucleic acid detecting cassette 100 according to an embodiment of the present invention is provided, wherein the nucleic acid detecting cassette 100 is used for detecting nucleic acid. The nucleic acid detecting cassette 100 includes a cassette body 1, a detecting chip 2, an electrophoresis cassette 3, and a connector 4. The detection chip 2 is disposed in the box 1, the detection chip 2 includes a first cover plate 21, a spacer layer 22 and a second cover plate 23, two opposite surfaces of the spacer layer 22 are respectively in contact with the first cover plate 21 and the second cover plate 23, the first cover plate 21, the spacer layer 22 and the second cover plate 23 enclose a channel 5, and the channel 5 is used for carrying a detection liquid a. The electrophoresis box 3 is disposed in the box body 1 and is communicated with the channel 5. The connector 4 is electrically connected to the detection chip 2 and the electrophoresis box 3, respectively, and the connector 4 is used for electrically connecting to an external control board. The nucleic acid detecting cassette 100 is used for performing nucleic acid amplification reaction and electrophoresis detection, a detecting liquid a containing a nucleic acid sample is added into a channel 5 of the detecting chip 2, it should be noted that the detecting liquid a exists in the channel 5 in the form of liquid beads, the detecting liquid a performs 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 cassette 3 from the detecting chip 2 for electrophoresis detection, and finally, an image of the electrophoresis cassette 3 is captured by an image collecting device matched with the nucleic acid detecting cassette 100, wherein the image is a fluorescence photograph of the electrophoresis detection. According to the invention, the detection chip 2 and the electrophoresis box 3 are integrated in the box body 1, the whole structure is simple, complex large-scale equipment is not needed, the cost is low, the detection liquid a can directly enter the electrophoresis box 3 for electrophoresis detection after completing nucleic acid amplification, the sample transfer matching connection process of different detection links is simplified, and the detection efficiency is improved.
Referring to fig. 1 to 5, the box body 1 includes a first housing 11, a second housing 12, a sample port 13 disposed on the second housing 12, and a detection window 14 disposed on the first housing 11. The first housing 11 and the second housing 12 together enclose a containing cavity (not shown), and the detecting chip 2, the electrophoresis cassette 3 and the connector 4 are all contained in the containing cavity. The sample addition port 13 is disposed corresponding to the detection chip 2, and is used for adding a detection solution a containing a nucleic acid sample into the detection chip 2. The detection window 14 is arranged corresponding to the electrophoresis box 3, and the image acquisition device can acquire the image of the electrophoresis box 3 through the detection window 14.
Referring to fig. 3, in the present embodiment, the first housing 11 and the second housing 12 are connected by a snap-fit manner, and after the first housing 11 and the second housing 12 are snapped together, the first housing 11 and the second housing 12 can be fastened by screws around the snap-fit manner, so as to increase the connection firmness of the first housing 11 and the second housing 12.
Referring to fig. 1 and 2, in the present embodiment, the sidewall of the box body 1 is further provided with an opening 17, the opening 17 is used for installing the connector 4, the connector 4 is integrally located in the accommodating cavity and is exposed out of the box body 1 through the opening 17, so that the connector 4 is conveniently electrically connected with an external control board.
Referring to fig. 2, in the embodiment, the cassette 1 further includes a card slot 15 disposed on the first housing 11, and since the nucleic acid detecting cassette 100 needs to be installed in the nucleic acid detecting apparatus during use, the card slot 15 is designed to facilitate the installation of the nucleic acid detecting cassette 100 in the used nucleic acid detecting apparatus.
Referring to FIG. 1, in conjunction with FIG. 13, in this embodiment, an indication mark 18 (for example, an arrow) is further disposed on a side of the second housing 12 away from the accommodating cavity, and referring to FIG. 13, the indication mark 18 is used for indicating an insertion direction of the nucleic acid detecting cassette 100 into the nucleic acid detecting apparatus 200, so as to avoid erroneous insertion.
Referring to fig. 3, in the present embodiment, a plurality of supporting structures 16 are disposed in the box body 1, and since the detecting chip 2, the electrophoresis box 3 and the connector 4 have different thicknesses in terms of structural design, the supporting structures 16 with different heights need to be designed to support the detecting chip 2, the electrophoresis box 3 and the connector 4 when being installed in the box body 1, so as to improve the connection stability between the detecting chip 2, the electrophoresis box 3 and the connector 4.
In this embodiment, the case 1 is made of plastic, and the supporting structure 16, the first housing 11 and the second housing 12 are integrally formed.
Referring to fig. 4 and 5, the detecting chip 2 further includes a driving circuit 24 disposed on a side of the first cover plate 21 close to the second cover plate 23, a first dielectric layer 26 disposed on a side of the driving circuit 24 close to the second cover plate 23, a conductive layer 25 disposed on a side of the second cover plate 23 close to the first cover plate 21, and a second dielectric layer 27 disposed on a side of the conductive layer 25 close to the first cover plate 21, wherein the driving circuit 24 and the conductive layer 25 are both electrically connected to the connector 4, and the detecting liquid a can move in the channel 5 according to a predetermined path by turning on or off the driving circuit 24 and the conductive layer 25.
Referring to fig. 5 and fig. 6 in combination with fig. 1, in the present 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, and the control electrode 242 is electrically connected to the connector 4. Specifically, the driving circuit 24 is a Thin Film Transistor (TFT) driving circuit, and since the detection liquid a has conductivity, the detection liquid a can move along a predetermined path in the channel 5 by combining with an electro wetting-On-Dielectric (EWOD) principle. Using the TFT principle, a circuit between a certain driving electrode 241 and the conductive layer 25 can be selectively turned on or off, so that the voltage between the driving electrode 241 and the conductive layer 25 is changed to change the wetting characteristics between the detection liquid a and the first and second dielectric layers 26 and 27, thereby controlling the detection liquid a to move along a predetermined path in the channel 5. As shown in fig. 5, the detection liquid a moves on the electrode I, the electrode H and the electrode G, and when the detection liquid a is on the electrode H, a voltage is applied between the electrode G and the conductive layer 25, a voltage Vd is applied to the electrode G, and the voltage between the electrode H and the conductive layer 25 is disconnected, at which time 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 liquid-solid contact angle between the electrode H and the detection liquid a becomes larger, and the liquid-solid contact angle between the electrode G and the detection liquid a becomes smaller, thereby urging the detection liquid a to move from the electrode H to the electrode G.
In this embodiment, the first dielectric layer 26 and the second dielectric layer 27 are both insulating hydrophobic layers, and may be specifically polytetrafluoroethylene coatings, which can play a role of insulating and hydrophobic on the one hand, and can also make the detection liquid a move more smoothly in a specified path on the other hand, thereby avoiding the liquid bead from breaking during the moving process.
In the present embodiment, referring to fig. 5, the driving circuit 24 is disposed on a side of the first cover plate 21 close to the channel 5. 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 side of the first cover plate 21 where the control electrode 242 is disposed is inserted into the connector 4 to electrically connect the detection chip 2 and the connector 4.
Referring to FIG. 6, in conjunction with FIG. 3 and FIG. 8, the driving circuit 24 can be divided into a plurality of regions, i.e., a sample application region A, a reagent storage region B, a plurality of nucleic acid amplification regions C, and a liquid discharge region D, according to different applications. The second cover plate 23 is provided with a first opening 29 corresponding to the sample addition region a, the first opening 29 corresponds to the sample addition port 13 of the cartridge body 1, and the detection solution a is added to the sample addition region a from the sample addition port 13 through the first opening 29. The reagent storage region B is used to store a fluorescent reagent (e.g., a fluorescent dye or a fluorescent probe). 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 a plurality of regions, and the number of the specific regions may be determined according to the actual detection requirement. The liquid outlet area D comprises a liquid outlet 51, the channel 5 is communicated with the electrophoresis cassette 3 through the liquid outlet 51, and the nucleic acid amplification product b can enter the electrophoresis cassette 3 through the liquid outlet 51 for electrophoresis detection in the liquid outlet area D.
Referring to fig. 5 and fig. 6 in combination with fig. 8, the specific moving path of the detection liquid a in the detection chip 2 is: after entering the sample addition region A, the detection solution a moves to the nucleic acid amplification region C according to a predetermined path under the drive of the drive electrode 241 to perform an amplification reaction; when the amplification reaction is completed, the amplified product moves to the reagent storage area B to be mixed with the fluorescent reagent, so that a nucleic acid amplification product B combined with the fluorescent reagent is obtained; the nucleic acid amplification product b is driven by the driving electrode 241 to move to the liquid outlet area D, and enters the electrophoresis cassette 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 the detection solution a can be subjected to different stages of nucleic acid amplification reactions at different temperatures.
In this embodiment, the fluorescent reagent is previously coated in the reagent storage region B during assembly of the detection chip 2, and it is not necessary to separately add the fluorescent reagent thereafter.
Referring to FIG. 3, in other embodiments, the fluorescent reagent may be mixed with the amplification product by subsequent addition. Specifically, a reagent tank 7 is disposed on the detection chip 2 corresponding to the reagent storage region B, and a fluorescent reagent can be added into the reagent tank 7 during nucleic acid detection.
Referring to fig. 3, 5 and 6, the detecting chip 2 further includes a heating element 28 disposed on a side of the first cover plate 21 and/or the second cover plate 23 away from the channel 5, wherein the heating element 28 is disposed corresponding to the nucleic acid amplification region C for heating the detecting liquid a. The heating element 28 is electrically connected to the connector 4, and a specific region of the channel 5 is heated by the heating element 28.
In the present embodiment, the heating element 28 is disposed on a side of the first cover plate 21 and the second cover plate 23 away from the channel 5.
In the present embodiment, the heating unit 28 is bonded to the surfaces of the first cover plate 21 and the second cover plate 23 by a heat conductive adhesive.
Referring to fig. 3 and 5, in the present 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 adhesive frame, and is adhered to the edges of the first cover plate 21 and the second cover plate 23 through the double-sided adhesive frame, so as to jointly form a sealed channel 5. Wherein the capacity of the channel 5 can be adjusted by designing spacer layers 22 with different thicknesses according to actual requirements.
In the present embodiment, after the detection chip 2 is assembled, silicone oil d is injected into the channel 5, and the detection liquid a moves through a predetermined path in the silicone oil d.
Referring to fig. 3, 4, 7, 8 and 11, the electrophoresis cassette 3 includes an electrophoresis tank 31, electrophoresis electrodes 32 disposed at two ends of the electrophoresis tank 31, a gel medium 33 disposed inside the electrophoresis tank 31, a liquid injection tank 34 disposed at one end of the gel medium 33, a connecting device 35 and a wetting liquid disposed in the electrophoresis tank 31. The electrophoresis electrode 32 is electrically connected to the connector 4, the electrophoresis cassette 3 further has a liquid inlet 37 corresponding to the liquid outlet 51, the connecting device 35 includes a first end 351 and a second end 352, the first end 351 extends into the channel 5 through the liquid inlet 37 and the liquid outlet 51, the second end 352 extends into the liquid injection groove 34, and the nucleic acid amplification product b enters the connecting device 35 through the liquid outlet 51 in the liquid outlet region D. And further into the liquid injection tank 34 of the gel medium 33, thereby performing the electrophoresis detection.
Referring to fig. 3, 4, 7 and 8, in the present embodiment, the electrophoresis tank 31 is located on a side of the first cover plate 21 away from the second cover plate 23, and an opening of the electrophoresis tank 31 faces a side of the first cover plate 21. The electrophoresis tank 31 comprises a transparent bottom plate 311 and a plurality of side walls 312 connected to the transparent bottom plate 311, wherein an end of the side wall 312 far away from the transparent bottom plate 311 contacts with a lower surface of the first cover plate 21, that is, the electrophoresis tank 31 utilizes the first cover plate 21 as electrophoresisThe cover plate of the tank 31 seals the electrophoresis cassette 3, and the liquid outlet 51 and the liquid inlet 37 belong to the same through hole. The above design makes a height difference deltaH between the silicone oil d in the detection chip 2 and the wetting liquid in the electrophoresis box 31The nucleic acid amplification product b in the channel 5 can smoothly enter the electrophoresis cassette 3; in addition, the structural design can improve the space utilization rate and is beneficial to reducing the volume of the whole nucleic acid detecting box 100. In this embodiment, a sealing rubber ring (not shown) is disposed between the sidewall 312 and the first cover plate 21 to improve the sealing performance of the electrophoresis cassette 3.
Referring to fig. 7, the electrophoresis tank 31 further includes a plurality of position-limiting blocks 313 disposed on the transparent bottom plate 311, the gel medium 33 is substantially a rectangular parallelepiped structure and can be limited between the position-limiting blocks 313, and the position-limiting blocks 313 can prevent the gel medium 33 from moving and deviating, thereby ensuring the accuracy of electrophoresis detection.
Referring to fig. 3, in the present embodiment, the transparent substrate 311 is a transparent glass plate, and the electrophoresis result can be observed.
In this embodiment, the number of the detents 313 is four, and the four detents 313 are located at four corners of the gel medium 33 having a rectangular parallelepiped structure, respectively, to fix the gel medium 33.
Referring to fig. 4 again, the electrophoresis tank 31 further includes a second opening 36, the second opening 36 is disposed at a position of the first cover plate 21 corresponding to the electrophoresis cassette 3, and a wetting fluid (e.g., buffer) can be injected into the electrophoresis tank 31 through the second opening 36.
Referring to fig. 8 to 10 in combination 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. Wherein the connecting means 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 cassette 3 by using the capillary effect. As shown in FIG. 8, in order to allow the nucleic acid amplification product b to smoothly enter the electrophoresis cassette 3, the end surface of the first end 351 needs to be flush with the surface of the silicone oil, i.e., the first end 351 includes a flat surface. Alternatively, as shown in fig. 9 and 10, the first end 351 is provided with at least one inclined surface, that is, the first end 351 corresponds to the connectionThe central axis c of the device 35 is arranged obliquely, in which case there is a step Δ H between the lowest point of the bevel and the lower surface of the channel 52The liquid surface of the silicone oil is located on the inclined surface, and the nucleic acid amplification product b can smoothly enter the connecting means 35. In the assembly design process of the connecting device 35 and the detecting chip 2, the connecting device 35 needs to be filled with a wetting liquid, and the wetting liquid needs to be capable of contacting with the liquid bead surface of the nucleic acid amplification product b at the liquid outlet region D to form a continuous liquid flow, so that the nucleic acid amplification product b can be ensured to smoothly enter the connecting device 35 by utilizing the capillary principle.
In this embodiment, the angle between the inclined plane and the central axis c of the connecting device 35 is 45 ° to 60 °, and experiments prove that the nucleic acid amplification product b can smoothly enter the connecting device 35 and then the gel medium 33 within this angle range.
In this embodiment, as shown in FIG. 9, the first end 351 of the connecting means 35 is formed with a slant having an inclination angle α of 45 ° to 60 °, and the nucleic acid amplification product b can smoothly enter the connecting means 35 and the gel medium 33 by the design of the slant and the capillary principle.
In another embodiment, as shown in FIG. 10, the connecting device 35 has two inclined planes with an inclination angle α of 45-60 degrees on opposite sides of the first end 351, and the nucleic acid amplification product b can smoothly enter the connecting device 35 and enter the gel medium 33 by the capillary principle through the design of the two inclined planes.
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.
Referring to FIG. 3, in conjunction with FIG. 8, since a height difference Δ H exists between the detecting chip 2 and the electrophoresis cassette 31In a normal stationary situation, the wetting fluid in the electrophoresis cartridge 3 will not enter the channel 5 through the connecting means 35. Since the surface of the first end 351 of the connecting means 35 is just flush with the surface of the silicone oil, the silicone oil in the channel 5 cannot enter the electrophoresis cassette 3 through the connecting means 35 in a stable condition. However, when the nucleic acid detecting cassette 100 is tilted, shaken or the internal pressure is changed during transportation, the nucleic acid detecting cassetteThe body can be simulated through high-altitude low pressure (0.2-0.7 bar) and vibration tests, and because of the pressure difference existing in the channel 5 and outside the channel 5 of the detection chip 2, and the pressure difference also existing inside and outside the electrophoresis box 3, the phenomenon that silicone oil in the channel 5 and wetting liquid in the electrophoresis box 3 leak or the mixture of the silicone oil and the wetting liquid occurs can be caused, the performance of the nucleic acid detection box 100 can be seriously influenced, and even the nucleic acid detection box 100 can be directly scrapped.
Referring to fig. 11 in conjunction with fig. 1, the present invention solves the above problem by adding a communicating structure 8 in order to avoid accidental leakage of silicone oil in the channel 5 and/or accidental mixing of the wetting fluid in the electrophoresis cassette 3 under the above special conditions. The communicating structure 8 is connected at one end to the first opening 29 and at the other end to the second opening 36 for communicating the channel 5 with the electrophoresis cassette 3, thereby equalizing the pressure in the channel 5 and the electrophoresis cassette 3 and avoiding accidental leakage of silicone oil in the channel 5 and the wetting fluid in the electrophoresis cassette 3 or accidental mixing of the two through the connecting means 35.
Referring to fig. 11, the communicating structure 8 includes a first connecting end 81 communicating with the first opening 29, a second connecting end 82 communicating with the second opening 36, and a connecting cavity 83 connecting the first connecting end 81 and the second connecting end 82. By connecting the communicating structure 8 between the first opening 29 and the second opening 36, communication between the channel 5 and the electrophoresis cassette 3 can be achieved, thereby balancing the pressure in the channel 5 and the electrophoresis cassette 3, which will press part of the silicone oil d into the connection cavity 83 via the first opening 29 when the pressure in the channel 5 is greater than the pressure in the electrophoresis cassette 3; when the pressure in the electrophoresis cassette 3 is higher than the pressure in the channel 5, a part of the wetting fluid will be squeezed via the second opening 36 into the connection chamber 83, thereby equalizing the pressure in the channel 5 and the electrophoresis cassette 3 and avoiding accidental leakage of silicone oil in the channel 5 and wetting fluid in the electrophoresis cassette 3 or accidental mixing of the two together via the connection means 35.
In this embodiment, referring to fig. 12 and fig. 11, the communicating structure 8 further includes a buffer cavity 84, and the buffer cavity 84 is disposed between the second connecting end 82 and the connecting cavity 83. The buffer chamber 84 is used for storing silicone oil d or a wetting fluid, and prevents the squeezed silicone oil d or wetting fluid from entering the electrophoresis cartridge 3 or the channel 5.
Referring to fig. 11, the electrophoresis box 3 is disposed below the detection chip 2, and the electrophoresis box 3 and the detection chip 2 are disposed in a staggered manner, and the projection area of the electrophoresis box 3 is larger than the projection area of the detection chip 2 along another direction perpendicular to the extending direction of the channel 5. The communication structure 8 further includes a first side wall 85, a second side wall 86, a third side wall 87, a bottom plate 88, and a top plate 89. The first sidewall 85 is disposed on the surface of the second cover plate 23, the second sidewall 86 and the third sidewall 87 are both disposed on the surface of the electrophoresis cassette 3 near the detection chip 2, the third sidewall 87 is disposed near the spacer 22, the second sidewall 86 is disposed far away from the spacer 22, the bottom plate 88 is disposed on the surface of the second cover plate 23 and connected to the third sidewall 87, and the top plate 89 is disposed at one end of the first sidewall 85 and the second sidewall 86 far away from the electrophoresis cassette 3. The first side wall 85, the second side wall 86, the third side wall 87, the bottom plate 88, the top plate 89 and the surface of the electrophoresis cartridge 3 close to the detection chip together enclose a cavity 9, the first opening 29 penetrates through the bottom plate 88 and is located in the cavity 9, the second opening 36 is located in the cavity 9, and the cavity 9 is respectively communicated with the channel 5 and the electrophoresis cartridge 3 through the first opening 29 and the second opening 36. The first connection end 81, the second connection end 82, the connection cavity 83 and the buffer cavity 84 together form the cavity 9.
In this embodiment, please refer to fig. 4 and fig. 11, the first cover plate 21 covers the opening of the electrophoresis tank 31, and the area of the first cover plate 21 is larger than the second cover plate 23. The first cover plate 21 extending beyond the second cover plate 23 can serve as the bottom of the communicating structure 8, and together with the first side wall 85, the second side wall 86, the third side wall 87, the bottom plate 88 and the top plate 89 form the cavity 9. In this manner, the assembly can be facilitated, and the volume of the electrophoresis cassette 3 can be reduced, which is advantageous for the miniaturization of the nucleic acid detecting cassette 100.
In this embodiment, referring to fig. 3 and fig. 11, the first opening 29 further penetrates the top plate 89 to communicate with the sample addition port 13, so as to add a sample to the sample addition region a.
In another embodiment, referring to fig. 13, the communicating structure 8a may also be a communicating pipe. One end of a communicating pipe is communicated with the first opening 29, and the other end is communicated with the second opening 36, so that the purpose of balancing the pressure in the channel 5 and the electrophoresis cassette 3 is achieved. And the communicating structure 8a is simple and convenient to assemble. If the communication tube made of rubber material is used, it can be bent in the nucleic acid detecting cassette 100 without occupying too much space.
Referring to FIG. 14, the present invention further provides a nucleic acid detecting apparatus 200, wherein the nucleic acid detecting apparatus 200 comprises a host 201 and the nucleic acid detecting cassette 100 as described above, the host 201 is provided with a detecting cassette mounting groove 202, and the nucleic acid detecting cassette 100 is mounted in the detecting cassette mounting groove 202.
Compared with the prior art, the nucleic acid detection kit provided by the invention integrates nucleic acid amplification reaction and electrophoresis detection, has a simple overall structure, is simple and convenient to detect and operate, has low professional requirements on the operation process, is high in detection efficiency, and greatly reduces the detection cost; meanwhile, the detection process has strong flexibility, does not need to be carried out in a fixed laboratory, and the nucleic acid detection box is portable and can realize community detection or family detection; the arrangement of the communicating structure can avoid the liquid mixing or leakage in the detection chip and the electrophoresis tank caused by the movement or vibration of the nucleic acid detection box, and improve the reliability of the nucleic acid detection box.
Claims (10)
1. A nucleic acid detecting cassette characterized by comprising:
the detection chip comprises a first cover plate, a spacing layer and a second cover plate, wherein two opposite surfaces of the spacing layer are respectively adjacent to the first cover plate and the second cover plate, the first cover plate, the spacing layer and the second cover plate are surrounded to form a channel, the first cover plate is provided with a liquid outlet, the second cover plate is provided with a first opening, the liquid outlet and the first opening are both communicated with the channel, and the channel is used for bearing detection liquid so that the detection liquid can perform nucleic acid amplification reaction in the channel to obtain a nucleic acid amplification product;
the electrophoresis box is arranged on one side, away from the channel, of the first cover plate and comprises a liquid inlet and a second opening, the liquid inlet is arranged corresponding to the channel, the second opening is arranged away from the channel, and the liquid outlet is communicated with the liquid inlet so that the nucleic acid amplification product enters the electrophoresis box; and
and the two ends of the communication structure are respectively communicated with the first opening and the second opening so as to communicate the channel with the electrophoresis box.
2. The nucleic acid detecting cassette according to claim 1, wherein the communicating structure includes a first connecting end communicating with the first opening, a second connecting end communicating with the second opening, and a connecting chamber connecting the first connecting end and the second connecting end.
3. The nucleic acid detecting cassette according to claim 2, wherein the communicating structure further comprises a buffer chamber provided between the second connecting end and the connecting chamber.
4. The nucleic acid detecting cassette according to claim 1, wherein the communicating structure is a communicating tube.
5. The nucleic acid detecting cassette according to claim 1, wherein the communicating structure includes a first side wall, a second side wall, a third side wall, a bottom plate, and a top plate, the first side wall is disposed on a surface of the second cover plate, the second side wall and the third side wall are both disposed on a surface of the electrophoresis cassette on a side close to the detection chip, the third side wall is disposed close to the spacer layer, the second side wall is disposed away from the spacer layer, the bottom plate is disposed on a surface of the second cover plate and connected to the third side wall, the top plate is disposed on an end of the first side wall and the second side wall away from the electrophoresis cassette, the first side wall, the second side wall, the third side wall, the bottom plate, the top plate, and a surface of the electrophoresis cassette on a side close to the detection chip together define a cavity, and the first opening extends through the bottom plate and is located in the cavity, the second opening is located in the cavity, and the cavity is communicated with the channel and the electrophoresis box through the first opening and the second opening respectively.
6. The nucleic acid detecting cassette according to claim 5, wherein the first opening is provided through the top plate.
7. The nucleic acid detecting cassette according to claim 1, wherein the electrophoresis cassette includes a connecting means having both ends connected to the liquid outlet and the liquid inlet, respectively, so that the channel communicates with the electrophoresis cassette, and the connecting means includes a first end extending into the channel via the liquid outlet.
8. The nucleic acid detecting cassette according to claim 7, wherein the electrophoresis cassette 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, and the connecting means includes a second end remote from the first end, the second end extending into the liquid injection tank.
9. The nucleic acid detecting cassette according to claim 8, wherein the first cover plate is provided at an opening of the electrophoresis tank.
10. A nucleic acid detecting apparatus characterized by comprising:
a host;
the detection box mounting groove is arranged on the host; and
the nucleic acid detecting cassette according to any of claims 1 to 9, which is detachably mounted in the cassette mounting groove.
Priority Applications (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP21199545.1A EP3978123A1 (en) | 2020-09-30 | 2021-09-28 | Nucleic acid detection kit and nucleic acid detection device |
| JP2021158319A JP2022058248A (en) | 2020-09-30 | 2021-09-28 | Nucleic acid detection kit and nucleic acid detection device |
| US17/488,647 US20220099620A1 (en) | 2020-09-30 | 2021-09-29 | Nucleic acid detection kit and nucleic acid detection device |
| PCT/CN2021/122431 WO2022068937A1 (en) | 2020-09-30 | 2021-09-30 | Nucleic acid testing device |
Applications Claiming Priority (6)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US202063085385P | 2020-09-30 | 2020-09-30 | |
| US202063085368P | 2020-09-30 | 2020-09-30 | |
| US63/085385 | 2020-09-30 | ||
| US63/085368 | 2020-09-30 | ||
| US202163146219P | 2021-02-05 | 2021-02-05 | |
| US63/146219 | 2021-02-05 |
Publications (1)
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| CN114317225A true CN114317225A (en) | 2022-04-12 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110746172.3A Pending CN114317225A (en) | 2020-09-30 | 2021-07-01 | Nucleic acid detecting cassette and nucleic acid detecting apparatus |
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| Country | Link |
|---|---|
| CN (1) | CN114317225A (en) |
| TW (1) | TW202214868A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101321867A (en) * | 2006-03-24 | 2008-12-10 | 株式会社东芝 | Nucleic acid detection cassette and nucleic acid detection system |
| WO2015111443A1 (en) * | 2014-01-27 | 2015-07-30 | 株式会社 日立ハイテクノロジーズ | Nucleic acid analyzing device |
| CN105277725A (en) * | 2014-07-01 | 2016-01-27 | 清华大学 | Integrated micro-fluidic system for nucleic acid analysis and detection |
| CN111592971A (en) * | 2020-07-07 | 2020-08-28 | 江苏汇先医药技术有限公司 | Micro-fluidic chip and method for nucleic acid detection |
-
2021
- 2021-07-01 CN CN202110746172.3A patent/CN114317225A/en active Pending
- 2021-07-01 TW TW110124306A patent/TW202214868A/en unknown
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101321867A (en) * | 2006-03-24 | 2008-12-10 | 株式会社东芝 | Nucleic acid detection cassette and nucleic acid detection system |
| WO2015111443A1 (en) * | 2014-01-27 | 2015-07-30 | 株式会社 日立ハイテクノロジーズ | Nucleic acid analyzing device |
| CN105277725A (en) * | 2014-07-01 | 2016-01-27 | 清华大学 | Integrated micro-fluidic system for nucleic acid analysis and detection |
| CN111592971A (en) * | 2020-07-07 | 2020-08-28 | 江苏汇先医药技术有限公司 | Micro-fluidic chip and method for nucleic acid detection |
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| TW202214868A (en) | 2022-04-16 |
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