CN113832014A - Nucleic acid detection device and detection method - Google Patents
Nucleic acid detection device and detection method Download PDFInfo
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- CN113832014A CN113832014A CN202111115363.6A CN202111115363A CN113832014A CN 113832014 A CN113832014 A CN 113832014A CN 202111115363 A CN202111115363 A CN 202111115363A CN 113832014 A CN113832014 A CN 113832014A
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Abstract
The invention provides a nucleic acid detection device and a detection method, comprising a cracking piece, a liquid transferring piece, an amplification piece and a rotating piece, wherein the cracking piece is provided with a cracking cavity, the liquid transferring piece is provided with a liquid transferring cavity, the amplification piece is provided with an amplification cavity, and the rotating piece can rotate to open or close the liquid transferring cavity. The nucleic acid detection device and the detection method provided by the invention have the advantages that the rotating part is controlled to rotate so as to seal the liquid transfer cavity, separate the liquid transfer cavity from the cracking cavity and the amplification cavity, open the liquid transfer cavity and communicate the cracking cavity with the liquid transfer cavity or communicate the liquid transfer cavity with the amplification cavity, the structure is simple, the operation is easy, and meanwhile, the quantitative transfer of liquid is realized through the liquid transfer cavity with constant volume.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a nucleic acid detection device and a detection method.
Background
Nucleic acid detection is an important detection mode for virus detection, and is implemented by extracting nucleic acid, carrying out nucleic acid amplification in a closed space and utilizingThe amplified nucleic acid reacts with a detection reagent to detect the virus. The loop-mediated isothermal amplification (LAMP) technology is to design 4-6 specific primers aiming at 6-8 regions of a target gene by using strand displacement DNA polymerase, and to realize 10 specific primers within dozens of minutes under the isothermal condition9-1010Techniques for secondary amplification. The loop-mediated isothermal amplification (LAMP) has the characteristics of high speed, simplicity, high sensitivity and specificity, has low requirements on instruments and experimental sites, and is widely applied to multiple fields of scientific research and nucleic acid detection. Cracking device, amplification device, detection device among the relevant nucleic acid detection device all set up alone, need frequently open cracking device, amplification device, detection device in the testing process in order to add and take out the operation, and the step is various, and the operation degree of difficulty is great, and inconvenient use, and increase the chance that sample liquid and external environment contacted easily, increase the contaminated possibility of sample liquid to reduce the precision of detecting.
Disclosure of Invention
In view of the above, the main objective of the embodiments of the present invention is to provide a nucleic acid detecting apparatus, so as to solve the technical problem of how to improve the detection accuracy by simple quantitative pipetting.
In order to achieve the above purpose, the technical solution of the embodiment of the present invention is realized as follows:
an embodiment of the present invention provides a nucleic acid detection apparatus, including: a cracking piece, which is internally provided with a cracking cavity; the pipetting piece is internally provided with a pipetting cavity, and the volume of the pipetting cavity is constant; an amplification member having an amplification chamber therein; a rotation member rotatable to close the pipetting chamber and open the pipetting chamber to communicate one of the lysis chamber and the amplification chamber.
In some embodiments, the pipetting chamber is in communication with the lysis chamber via a first channel and the pipetting chamber is in communication with the amplification chamber via a second channel.
In some embodiments, the rotation member comprises a first rotation member at least partially positioned within the lysis chamber and configured to rotate to open and close the first channel, and a second rotation member at least partially positioned within the amplification chamber and configured to rotate to open and close the second channel.
In some embodiments, the first rotating member includes a first rotating disc, a first supporting member and a second rotating disc sequentially arranged along a first direction, and two opposite ends of the first supporting member are respectively connected with the first rotating disc and the second rotating disc; the second rotating disc is positioned in the cracking cavity, a first opening penetrating through the second rotating disc along the first direction is formed in the second rotating disc, and the first opening is communicated with or separated from the first channel along with the rotation of the first rotating part.
In some embodiments, an end of the first carousel distal to the second carousel protrudes out of the lysis chamber.
In some embodiments, the first rotating disk is provided with a second opening penetrating through the first rotating disk along the first direction.
In some embodiments, the second rotating part includes a third rotating disc, a second supporting part and a fourth rotating disc sequentially arranged along the first direction, and two opposite ends of the second supporting part are respectively connected with the third rotating disc and the fourth rotating disc; the third rotating disc is positioned in the amplification cavity, a third opening penetrating through the third rotating disc along the first direction is formed in the third rotating disc, and the third opening is communicated with or separated from the second channel along with the rotation of the second rotating element.
In some embodiments, an end of the fourth rotating disk distal to the third rotating disk protrudes out of the amplification chamber.
In some embodiments, the lysis member, the pipetting member and the amplification member are connected in series in a first direction.
In some embodiments, the first rotating member and the second rotating member are provided with sealing rings to seal the lysis chamber and the amplification chamber, respectively.
An embodiment of the present invention further provides a detection method for any one of the above nucleic acid detection apparatuses, including: s1, the rotating piece rotates to communicate the pipetting cavity with the lysis cavity and separate the pipetting cavity from the amplification cavity; s2, the pipetting cavity acquires a set amount of liquid entering from the lysis cavity; s3, the rotating piece rotates to separate the liquid transferring cavity from the cracking cavity and communicate the liquid transferring cavity with the amplification cavity; s4, the amplification chamber obtains the set amount of liquid entering from the liquid transferring chamber.
In some embodiments, before the step S1, the method further includes: and S0, adjusting the rotating piece to be at an initial position, wherein the initial position is a position at which the pipetting cavity is separated from the lysis cavity and the amplification cavity.
The nucleic acid detection device and the detection method provided by the invention comprise a cracking piece, a liquid transfer piece, an amplification piece and a rotating piece, wherein the cracking piece is provided with a cracking cavity, the liquid transfer piece is provided with a liquid transfer cavity, the amplification piece is provided with an amplification cavity, and the rotating piece can rotate to open or close the liquid transfer cavity. The nucleic acid detection device and the detection method provided by the invention have the advantages that the rotating part is controlled to rotate so as to seal the liquid transfer cavity, separate the liquid transfer cavity from the cracking cavity and the amplification cavity, open the liquid transfer cavity and communicate the cracking cavity with the liquid transfer cavity or communicate the liquid transfer cavity with the amplification cavity, the structure is simple, the operation is easy, and meanwhile, the quantitative transfer of liquid is realized through the liquid transfer cavity with constant volume.
Drawings
FIG. 1 is a sectional view of a nucleic acid detecting apparatus according to an embodiment of the present invention;
FIG. 2 is a sectional view of another nucleic acid detecting apparatus according to an embodiment of the present invention;
fig. 3 is a schematic structural diagram of a first rotating member according to an embodiment of the present invention;
fig. 4 is a schematic structural diagram of a second rotating member according to an embodiment of the present invention;
FIG. 5 is an exploded view of a nucleic acid detecting apparatus according to an embodiment of the present invention;
FIG. 6 is a schematic view of an assembly of a lysis member and a pipetting member according to an embodiment of the invention;
FIG. 7 is an exploded view of another nucleic acid detecting apparatus according to an embodiment of the present invention.
Description of reference numerals:
10. a cracking member; 11. a lysis chamber; 20. a pipetting member; 21. a pipetting chamber; 22. a first through hole; 23. a second through hole; 24. a first channel; 25. a second channel; 30. an amplification element; 31. an amplification chamber; 40. a rotating member; 401. a communicating hole; 41. a first rotating member; 411. a first turntable; 4111. a second opening; 412. a first support member; 413. a second turntable; 4131. a first opening; 42. a second rotating member; 421. a third turntable; 4211. a third opening; 422. a second support member; 423. a fourth turntable; 50. and (5) sealing rings.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The individual features described in the embodiments can be combined in any suitable manner without departing from the scope, for example different embodiments and aspects can be formed by combining different features. In order to avoid unnecessary repetition, various possible combinations of the specific features of the invention will not be described further.
In the following description, the terms "first \ second \ third \ fourth \ are used merely to distinguish different objects and do not indicate that the objects have the same or relative relationship. It should be understood that the description of the "upper", "lower" and "inner" directions as the directions in the normal use state, the "upper" and "lower" directions indicate the up-down direction indicated in the corresponding schematic diagram, and may or may not be the left-right direction in the normal use state, and the "first direction" is the up-down direction indicated in the corresponding schematic diagram.
It should be noted that the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other like elements in a process, method, article, or apparatus that comprises the element. The term "coupled", where not otherwise specified, includes both direct and indirect connections.
The nucleic acid detection device provided by the invention can detect nucleic acid in a short time, and is mainly applied to the scene of on-site instant detection, namely on-site instant detection, which mainly means that rapid detection is carried out on the site outside a laboratory.
The following is an exemplary illustration of the nucleic acid detection process:
in the nucleic acid detection, after a detection sample is obtained by a detector, the detection sample is placed into a cracking container for cracking reaction, cell lysate can be placed in the cracking container, the detection sample is contacted with the cell lysate and reacts, a cell membrane of the detection sample is dissolved, and cell contents are released, wherein the cell contents comprise nucleic acid, protein and other substances. Thereafter, the solution containing the nucleic acid is transferred to an amplification vessel, and the nucleic acid is subjected to amplification detection. In the above detection process, quantitative transfer of liquid is required to be realized, so as to obtain an accurate detection result.
In the embodiment of the present invention, as shown in FIG. 1, the nucleic acid detecting apparatus 1 includes a lysis member 10, a pipetting member 20, a detection member 30, and a rotation member 40.
As shown in fig. 1, the lysis member 10 has a lysis chamber 11 inside. The lysis member 10 can provide a reaction space for the sample to lyse and release the cell content, i.e. a lysis chamber 11, wherein the lysis chamber 11 is a hollow chamber and can contain the reactant. In some embodiments, the temperature of the cleavage member 10 can be controlled depending on the reaction temperature requirements. For example, in an environment where pyrolysis is desired, a heating plate may be attached to the pyrolysis member 10 to raise the temperature within the pyrolysis chamber 11. The temperature inside the lysis chamber 11 can also be maintained by placing the lysis member 10 in a constant temperature liquid so that the substances in the lysis chamber 11 react at a constant temperature.
As shown in FIG. 1, the pipetting member 20 has a pipetting chamber 21 therein, and the pipetting chamber 21 has a constant volume. Specifically, the pipetting device 20 is used for receiving the liquid after the completion of the lysis reaction in the lysis device 10, wherein the volume of the pipetting cavity 21 is constant and is smaller than the volume of the lysis cavity 11, so that the pipetting cavity 21 can be filled each time the liquid in the lysis cavity 11 is transferred into the pipetting cavity 21, and thus the amount of the liquid transferred from the lysis cavity 11 is the same amount each time, and quantitative pipetting is realized.
As shown in FIG. 1, the amplification member 30 has an amplification chamber 31 inside. Specifically, the amplification chamber 31 is also a hollow chamber and can contain reactants. The volume in the amplification chamber 31 is at least larger than the volume of the pipetting chamber 21, so that the liquid in the pipetting chamber 21 can be completely transferred to the detection chamber 31, and the liquid transferred from the pipetting chamber 21 to the amplification chamber 31 is the same each time, thereby realizing quantitative pipetting. After the liquid is transferred to the amplification chamber 31, an amplification reaction can be performed to amplify the nucleic acid and increase the content of the nucleic acid, for example, a primer hexamer, phi 29DNA polymerase, deoxyinosine-like primer, etc. can be added to the amplification chamber 31 to promote the amplification of the nucleic acid. Optionally, a detection reagent may be disposed in the amplification chamber 31, so that the nucleic acid reacts with the detection reagent, thereby achieving the purpose of completing the amplification and detection of the nucleic acid by using only one amplification chamber 31. In some embodiments, the temperature of the amplification member 30 can be controlled according to the reaction temperature requirements. For example, in an environment where high temperature amplification is required, a heating plate may be attached to the amplification member 30 to raise the temperature in the amplification chamber 31. The amplification member 30 may be placed in a constant temperature liquid to maintain the temperature in the amplification chamber 31, so that the substances in the lysis chamber 11 react at a constant temperature.
In some embodiments, the lysis member 10, the pipetting member 20 and the amplification member 30 may be integrally formed as one integral device, may each be formed as separate devices, or may be formed as one integral device for each two. For example, the lysis member 12 may be formed as an integral device with the pipetting member 20, wherein the lysis chamber 11 and the pipetting chamber 21 are spaced apart and the amplification member 30 is formed as a separate device. As shown in FIG. 1, the rotation member 40 is rotatable to close the pipetting chamber 21 and open the pipetting chamber 21 to communicate one of the lysis chamber 11 and the amplification chamber 31. Specifically, the rotating member 40 is a rotatable member, and the opening and closing of the pipetting cavity 21 can be controlled by controlling the rotating member 40, so as to control the communication between the pipetting cavity 21 and the lysis cavity 11 or the communication cavity 21.
In some embodiments, as shown in FIG. 1, one end of the rotating member 40 protrudes into the pipetting cavity 21 and the other end protrudes out of the pipetting cavity 21 and out of the pipetting member 20, and the portion of the rotating member 40 protruding out of the pipetting member 20 can be used to detect a person manually rotating the rotating member 40. The part of the rotating member 40 extending into the liquid transferring cavity 21 is provided with a communicating hole 401, the communicating hole 401 rotates along with the rotating member 40 to communicate the liquid transferring cavity 21 with the lysis cavity 11 or the liquid transferring cavity 21 with the amplification cavity 31, the communicating hole 401 can be a strip-shaped through hole, wherein the rotating member 40 does not fill the liquid transferring cavity 21 but leaves a certain space to contain liquid, one end of the communicating hole 401 is communicated with the space to contain liquid, and the other end can be communicated with the lysis cavity 11 or the amplification cavity 31. The pipetting member 20 can be provided with a second through hole 23, the first through hole 22 is communicated with the lysis chamber 11, and the second through hole 23 is communicated with the amplification chamber 31. The rotating member 40 can rotate, when the connecting hole 401 is communicated with the first through hole 22, the liquid in the lysis chamber 11 can enter the connecting hole 401 through the first through hole 22 and then flow into the liquid containing space in the liquid moving chamber 21 through the connecting hole 401; when the connection hole 401 communicates with the second through hole 23, the liquid in the pipetting cavity 21 can enter the connection hole 401 through the second through hole 23 and then flow into the amplification cavity 31 through the connection hole 401.
Alternatively, the lysis member 10, the pipetting member 20, the amplification member 30 and the rotation member 40 can be transparent, so that the detection personnel can directly observe the transfer liquid in the lysis chamber 11, the pipetting chamber 21 and the amplification chamber 31 and the reaction condition.
The nucleic acid detection device provided by the embodiment of the invention comprises a cracking piece, a liquid transfer piece, an amplification piece and a rotating piece, wherein the cracking piece is provided with a cracking cavity, the liquid transfer piece is provided with a liquid transfer cavity, the amplification piece is provided with an amplification cavity, and the rotating piece can rotate to open or close the liquid transfer cavity. The nucleic acid detection device provided by the embodiment of the invention has the advantages that the rotating part is controlled to rotate so as to seal the liquid transfer cavity, separate the liquid transfer cavity from the cracking cavity and the amplification cavity, open the liquid transfer cavity and communicate the cracking cavity with the liquid transfer cavity or communicate the liquid transfer cavity with the amplification cavity, the structure is simple, the operation is easy, and meanwhile, the quantitative transfer of liquid is realized through the liquid transfer cavity with constant volume.
In some embodiments, as shown in FIG. 1, the pipetting chamber 21 communicates with the lysis chamber 11 via a first channel 24 and the pipetting chamber 21 communicates with the amplification chamber 31 via a second channel 25. The first channel 24 is a tube with two open ends, one end of the first channel 24 is communicated with the lysis chamber 11, and the other end of the first channel 24 is communicated with the first through hole 22 of the pipetting chamber 21, so that the liquid in the lysis chamber 11 can be transferred to the pipetting chamber 21 through the first channel 24. The second channel 25 is a tube with two open ends, one end of the second channel 25 is communicated with the amplification part 30, and the other end of the second channel 25 is communicated with the pipetting cavity 21, so that the liquid in the pipetting cavity 21 can be transferred to the amplification cavity 31 through the second channel 25. The first channel 24 is arranged between the lysis cavity 11 and the pipetting cavity 21, the second channel 25 is arranged between the pipetting cavity 21 and the amplification cavity 31, the lysis cavity 11, the pipetting cavity 21 and the amplification cavity 31 are arranged at intervals, and therefore heat in the lysis cavity 11, the pipetting cavity 21 and the amplification cavity 31 is isolated, and the heat insulation effect is achieved.
In some embodiments, as shown in fig. 2, the rotating member 40 includes a first rotating member 41 and a second rotating member 42. The rotating member 40 can control the closing of the pipetting chamber 21 and the communication of the pipetting chamber 21 with the lysis chamber 11 or the amplification chamber 31 by controlling the first rotating member 41 and the second rotating member 42, respectively. The first rotating member 41 is at least partially located in the lysis chamber 11 and rotates to open and close the first channel 24. The first rotating member 41 is disposed in the lysis chamber 11 and adjacent to an opening at one end of the first channel 24, and by rotating the first rotating member 41, the opening can be closed to prevent the liquid in the lysis chamber 11 from entering the first channel 24, or the first rotating member 41 can be rotated to change the lysis chamber 11 and the first channel 24 from a closed state to a communicated state, so that the liquid in the lysis chamber 11 can be transferred to the liquid-transferring chamber 21 through the first channel 24. The first rotating member 41 may be disposed in the first cracking chamber 11, and extend into the cracking chamber 11 through an external device to be connected to the first rotating member 41, and provide a rotational driving force to drive the first rotating member 41 to rotate, for example, a cross-shaped connection port is disposed on the first rotating member 41, a cross-shaped rotation shaft corresponding to the external device is disposed on the external device, a through hole for passing the rotation shaft is disposed on the cracking member 11, the rotation shaft is connected to the first rotating member 41 through the through hole, and the external device drives the rotation shaft to rotate, so as to drive the first rotating member 41 to rotate. The first rotating member 41 may also partially extend into the cracking cavity 11 to seal or communicate the cracking cavity 11 and the first channel 24, and the other part of the first rotating member 41 extends out of the cracking cavity 11, and the external device may rotate the part of the first rotating member 41 extending out of the cracking cavity 11 to drive the part of the first rotating member 41 located in the cracking cavity 11, thereby realizing the partition or communication between the cracking cavity 11 and the first channel 24. It should be noted that the first rotating member 40 is located in the lysis chamber 11, and occupies a part of the space in the lysis chamber 11, so the space for the sample to undergo the lysis reaction is the whole space of the lysis chamber 11 except the space occupied by the first rotating member 40.
As shown in FIG. 2, a second rotatable member 42 is at least partially positioned within the amplification chamber 31 and is rotated to open and close the second channel 25. The second rotating member 42 is disposed in the amplification chamber 31 adjacent to an opening at one end of the second channel 25, and the opening can be closed by rotating the second rotating member 42 so that the liquid in the pipetting chamber 21 cannot enter the second channel 25, or the second rotating member 42 is rotated so that the pipetting chamber 21 and the second channel 25 are changed from a closed state to a communicated state so that the liquid in the pipetting chamber 21 can be transferred to the amplification chamber 31 through the second channel 25. The second rotating member 42 may be disposed in the amplification chamber 31, and extend into the amplification chamber 31 through an external device to connect with the second rotating member 42, and provide a rotational driving force to drive the second rotating member 42 to rotate, for example, a cross-shaped connection port is disposed on the second rotating member 42, a cross-shaped rotation shaft corresponding to the cross-shaped connection port is disposed on the external device, a through hole for passing the rotation shaft is disposed on the amplification member 31, the rotation shaft is connected with the second rotating member 42 through the through hole, and the external device drives the rotation shaft to rotate, so as to drive the second rotating member 42 to rotate. The second rotating member 42 can also partially extend into the amplification chamber 31 to seal or communicate the amplification chamber 31 and the second channel 23, another part of the second rotating member 42 extends out of the amplification chamber 31, and the external device can drive a part of the second rotating member 42 located in the amplification chamber 31 to rotate by rotating the part of the second rotating member 42 extending out of the amplification chamber 31, thereby realizing the isolation or communication between the amplification chamber 31 and the second channel 25. It should be noted that the second rotating member 42 is disposed in the amplification chamber 31 and occupies a part of the space in the amplification chamber 31, and thus the space in the amplification chamber 31 where the liquid is subjected to the amplification reaction and the detection reaction is the entire space of the amplification chamber 31 minus the space occupied by the second rotating member 42.
Through setting up first passageway intercommunication schizolysis chamber and move liquid chamber, through the setting of first rotating part in the schizolysis chamber to the first passageway of switch, thereby realize that first rotating part is rotatory to be connected or cut off schizolysis chamber and move liquid chamber. The second channel is arranged to be communicated with the liquid transferring cavity and the amplification cavity, and the second rotating part is arranged in the amplification cavity to open and close the second channel, so that the second rotating part is rotatably communicated with or separates the liquid transferring cavity and the amplification cavity.
In some embodiments, as shown in fig. 2 and 3 in combination, the first rotating member 41 includes a first rotating disk 411, a first supporting member 412 and a second rotating disk 413, which are sequentially arranged along the first direction, and two opposite ends of the first supporting member 412 are respectively connected to the first rotating disk 411 and the second rotating disk 413. Specifically, the first direction is a direction in which the length of the first support 412 extends, i.e., a direction from top to bottom in the drawing. The first rotating disk 411 and the second rotating disk 413 are substantially disc-shaped, the first supporting member 412 is a rod-shaped member, one end of the first supporting member 412 is connected with the center of the first rotating disk 411, the other end of the first supporting member 412 is connected with the center of the second rotating disk 413, and the first rotating disk 411 and the second rotating disk 413 are supported by the first supporting member 412, so that the first rotating disk 411 and the second rotating disk 413 are arranged at intervals in the extending direction of the first supporting member 412. The second rotating disk 413 is located in the cracking chamber 11, and the second rotating disk 413 is provided with a first opening 4131 penetrating through the second rotating disk 413 along the first direction, and the first opening 4131 is communicated with or isolated from the first passage 24 along with the rotation of the first rotating member 41. Specifically, the lower end of the pyrolysis chamber 11 in the first direction may communicate with the opening of the first passage 24, and the second rotating disk 413 may contact the lower end of the pyrolysis chamber 11 and may rotate with respect to the lower end of the pyrolysis chamber 11. First opening 4131 of second disk 413 is rotated with second disk 413 to oppose or offset the opening of first passage 24, e.g., second disk 413 is rotated such that first opening 4131 opposes the opening of first passage 24, thereby allowing lysis chamber 11 to communicate with first passage 24, and if second disk 413 is rotated such that first opening 4131 is offset from the opening of first passage 24, the other portion of second disk 413 covers the opening of first passage 24, thereby isolating first passage 24 and lysis chamber 11. It should be noted that the first opening 4131 and the opening of the first channel 24 are not required to be completely opposite, but only partially opposite to each other so that the liquid can pass through to communicate the lysis chamber 11 and the first channel 24.
Alternatively, the first opening 4131 may be configured as a sector-shaped opening, the sector-shaped opening is concentric with the second rotating disk 413, and the first passage 24 may also be configured as a corresponding sector-shaped structure, so that the first rotating member 41 drives the second rotating disk 413 to rotate, and the communication area between the first opening 4131 and the first passage 24 may be controlled by the rotation angle of the second rotating disk 413, so as to control the liquid flow rate.
In some embodiments, as shown in conjunction with fig. 2 and 3, an end of first rotating disk 411 distal to second rotating disk 413 protrudes out of lysis chamber 11. Specifically, the first rotating disc 411 protrudes out of the lysis chamber 11 in a first direction from top to bottom in the figure, and the portion of the first rotating disc 411 protruding out of the lysis chamber 11 can be used for the detection person to manually rotate the first rotating member 41. The first rotating disc 411 is forced to rotate to drive the second supporting member 412 to rotate, and the first supporting member 412 rotates to drive the second rotating disc 413 to rotate, so that the first opening 4131 can connect or separate the lysis chamber 11 and the pipetting chamber 21, thereby facilitating the pipetting operation.
In some embodiments, as shown in fig. 2 and 3, the first rotating disk 411 is provided with a second opening 4111 extending through the first rotating disk 411 along the first direction. Specifically, the first rotary disc 411 seals the opening of the lysis chamber 11 communicating with the external environment to prevent the liquid leakage from the lysis chamber 11, and the second opening 4111 is provided on the first rotary disc 411 to facilitate the sample to be put into the lysis chamber 11. Optionally, a sealing film may be disposed on the second opening 4111, and after the sample is placed in the cracking chamber 11, the sealing film is used to seal the second opening 4111 to prevent the cracking chamber 11 from communicating with the external environment, so that the reaction in the cracking chamber 11 is completed in a sealed environment. A closing cover may also be disposed on the first rotating disk 411, and the closing cover is used to cover the second opening 4111. The closing cover is opened to place the sample in the cracking cavity 11, and the closing cover can cut off the communication between the cracking cavity 11 and the external environment.
In some embodiments, as shown in fig. 2 and 4, the second rotating member 42 includes a third rotating disc 421, a second supporting member 422, and a fourth rotating disc 423 sequentially arranged along the first direction, and opposite ends of the second supporting member 422 are respectively connected to the third rotating disc 421 and the fourth rotating disc 4243. Specifically, the first direction is a direction in which the length of the second support 422 extends, i.e., a direction from top to bottom in the drawing. The third rotating disc 421 and the fourth rotating disc 423 are substantially disc-shaped, the second supporting member 422 is a rod-shaped member, one end of the second supporting member 422 is connected to a circle center of the third rotating disc 421, the other end of the second supporting member 422 is connected to a circle center of the fourth rotating disc 423, and the third rotating disc 421 and the fourth rotating disc 423 are supported by the second supporting member 423, so that the third rotating disc 421 and the fourth rotating disc 423 are arranged at intervals in an extending direction of the second supporting member 422. The third rotating disc 421 is located in the amplification chamber 31, and the third rotating disc 421 is provided with a third opening 4211 penetrating through the third rotating disc 421 along the first direction, and the third opening 4211 is communicated with or isolated from the second channel 25 along with the rotation of the second rotating member 42. Specifically, the lower end of the amplification chamber 31 in the first direction may communicate with the opening of the second channel 25, and the third rotating disk 421 may contact the lower end of the amplification chamber 31 and may rotate with respect to the lower end of the amplification chamber 31. For example, when the third dial 421 is rotated to make the third opening 4211 face the opening of the second passage 25, the amplification chamber 31 can communicate with the second passage 25, and when the third dial 4211 is rotated to make the third opening 4211 face the opening of the second passage 25, the other part of the third dial 4211 covers the opening of the second passage 25, thereby blocking the second passage 25 and the amplification chamber 31. It should be noted that the third opening 4211 and the opening of the second channel 25 are not required to be completely opposite, but only partially opposite to each other so that the liquid can pass through the amplification chamber 31 and the second channel 25.
Alternatively, the third opening 4211 may be a sector opening, which is concentric with the third turntable 421, and the second passage 25 may also be a corresponding sector structure, so that the second rotating member 42 drives the third turntable 421 to rotate, and the communication area between the third opening 4211 and the second passage 25 may be controlled by the rotation angle of the third turntable 421, so as to control the flow rate of the liquid.
In some embodiments, as shown in conjunction with fig. 2 and 4, an end of the fourth rotating disk 423 remote from the third rotating disk 421 protrudes out of the amplification chamber 31. Specifically, the fourth rotating disc 423 protrudes upward from the amplification chamber 31 in the first direction (the up-down direction shown in fig. 5), and the protruding portion of the fourth rotating disc 423 from the amplification chamber 31 can be used for the inspector to manually rotate the second rotating member 42. The fourth rotating disc 423 is forced to rotate to drive the second supporting member 422 to rotate, and the second supporting member 422 rotates to drive the third rotating disc 421 to rotate, so that the third opening 4211 can communicate or separate the pipetting cavity 21 and the amplification cavity 31, thereby facilitating pipetting operation.
In some embodiments, as shown in FIG. 5, the lysis member 10, the pipetting member 20 and the amplification member 30 are connected in series in a first direction. Specifically, the first direction may be a vertical direction as shown in FIG. 6, and the lysis member 10, the pipetting member 20 and the amplification member 30 are connected in sequence to form a straight line, so that the lysis chamber 11 is located above the pipetting chamber 21, and the pipetting chamber 21 is located above the amplification chamber 31, which can make the whole nucleic acid detecting apparatus compact and save materials. Under the action of gravity, the liquid in the lysis chamber 11 can flow into the pipetting chamber 21, and the liquid in the pipetting chamber 21 can also flow into the amplification chamber 31, so that the liquid can be transferred without using a pipetting pump or the like.
Alternatively, the walls of the lysis chamber 11 and the pipetting chamber 21 may be provided with a plurality of hydrophobic grooves extending in the up-down direction shown in fig. 7, and the hydrophobic grooves may be provided around the walls of the chambers to assist in the transfer of the liquid in the lysis chamber 11 and the pipetting chamber 21.
In some embodiments, as shown in FIG. 5, the first rotating member 41 and the second rotating member 42 are provided with sealing rings 50 for sealing the lysis chamber 11 and the amplification chamber 31, respectively. Specifically, the sealing rings 50 may be of a circular ring structure, the sealing rings 50 are provided with a plurality of sealing rings, and are respectively located in the cracking chamber 11 and the amplification chamber 31, for example, the sealing rings 50 located in the cracking chamber 11, wherein a certain number of the sealing rings are provided between the second rotating disc 413 and the chamber wall of the cracking chamber 11, and are simultaneously in contact with the second rotating disc 413 and the chamber wall of the cracking chamber 11, and the other sealing rings 50 are provided between the first rotating disc 411 and the chamber wall of the cracking chamber 11, and are simultaneously in contact with the first rotating disc 411 and the chamber wall of the cracking chamber 11, so as to seal gaps between the first rotating disc 411 and the second rotating disc 413 and the chamber wall of the cracking chamber 11, and avoid liquid leakage in the cracking chamber 11. A certain number of sealing rings 50 are arranged in the amplification chamber 31 between the third rotating disc 421 and the wall of the amplification chamber 31 and are simultaneously in contact with the third rotating disc 421 and the wall of the amplification chamber 31, and other sealing rings 50 are arranged between the fourth rotating disc 423 and the wall of the amplification chamber 31 and are simultaneously in contact with the fourth rotating disc 423 and the wall of the detection chamber 31, so that the gaps between the third rotating disc 421 and the fourth rotating disc 423 and the wall of the amplification chamber 31 are sealed, and the liquid in the amplification chamber 31 is prevented from leaking.
In addition to the structure of the first rotating member 41 and the second rotating member 42 in any one of the above embodiments, in other embodiments, the first rotating member 41 and the second rotating member 42 may also be a rotary pick, and the structure of the first rotating member 41 and the second rotating member 42 will be exemplarily described below with reference to fig. 6 and 7.
As shown in fig. 6, the first rotating member 41 can be a rotating shifting piece, and is disposed between the lysis member 10 and the pipetting member 20, wherein both the lysis chamber 11 and the pipetting chamber 21 are opened with openings communicating with the outside, and the openings of both the lysis chamber 11 and the pipetting chamber 21 are opposite to communicate the lysis chamber 11 and the pipetting chamber 21. The rotary paddle is rotatable about its axis to block completely the opening of the lysis chamber 11 opposite the pipetting chamber 21 or to block partially the opening of the lysis chamber 11 opposite the pipetting chamber. The rotary shifting piece can be rotated automatically or manually, for example, the rotary shifting piece can be driven by a motor which can be arranged on the rotation center of the rotary shifting piece, so that the rotary shifting piece can rotate around the rotation center at regular intervals for periodic rotation, for example, the rotary shifting piece rotates once every 60 minutes, and the time for rotating the rotary shifting piece 60 once can be 1 minute, so that the liquid in the lysis cavity 11 can be filled in the liquid transferring cavity 21. The rotary shifting piece can also provide rotary driving force through the rotary rod, one end of the rotary rod is connected with the rotary center of the rotary shifting piece, and the other end of the rotary rod can penetrate through the cracking cavity 11 and protrude out of the cracking piece 10, so that the rotary rod can be manually rotated by a detection person, and the rotary shifting piece is driven to rotate. The rotating shifting piece completely blocks the orifice of the cracking cavity 11 opposite to the liquid transferring cavity 21 to cut off the communication between the cracking cavity 11 and the liquid transferring cavity 21, so that the liquid in the cracking cavity 11 is prevented from being transferred to the liquid transferring cavity 21, and the rotating shifting piece partially shields the orifice of the cracking cavity 11 opposite to the liquid transferring cavity. So that the mouths of both the lysis chamber 11 and the pipetting chamber 21 can be communicated, and the liquid in the lysis chamber 11 can be transferred into the pipetting chamber 21.
The second rotating member 42 is also a rotatable member, and the communication or the disconnection of the amplification chamber 31 and the pipetting chamber 21 is controlled by rotating the second rotating member 42. For example, the second rotating member 42 may be a rotary paddle at least partially located in the amplification chamber 31, and the structure of the rotary paddle may be the same as that of the rotary paddle used in the first rotating member 41, which will not be described herein.
It should be noted that the first rotating member 41 and the second rotating member 42 may have other structures. The first rotating member 44 and the second rotating member 42 may have the same or different structures, and the first rotating member 41 and the second rotating member 42 are relatively independent from each other, and the rotation states of the two members do not interfere with each other. For example, as shown in fig. 7, the first rotating member 41 and the second rotating member 42 are both rotary paddles, and both rotary paddles can rotate to control the opening and closing of the pipetting chamber 21. When the first rotating member 41 and the second rotating member 42 each cover the opening of the pipetting chamber 21, the pipetting chamber 21 is closed so as not to communicate with the lysis chamber 11 and the amplification chamber 31. When the first rotating member 41 rotates, the first passage 24 is at least partially uncovered, so that the pipetting cavity 24 communicates with the lysis chamber 11 through the first passage 24. When the second rotating member 42 rotates, the second channel 25 is at least partially uncovered, so that the pipetting chamber 21 communicates with the amplification chamber 31 through the second channel 25. For another example, one of the first rotating member 41 and the second rotating member 42 may be a structure with a rotating disk in the above-mentioned still another embodiment, and the other may be a structure provided as a rotary paddle.
An embodiment of the present invention further provides a detection method for a nucleic acid detection device in any one of the above embodiments, including:
s1, the rotating member 40 rotates to connect the pipetting cavity 21 and the lysis cavity 11 and separate the pipetting cavity 21 and the amplification cavity 31.
The rotating member 40 can rotate to control the connection and disconnection between the pipetting cavity 21 and the lysis cavity 11, for example, the rotating member 40 rotates by a certain extent to connect the pipetting cavity 21 and the lysis cavity 11, and then the rotating member 40 rotates to a certain position again to connect the pipetting cavity 21 and the lysis cavity 11. When nucleic acid detection is performed, a detection sample can be added into the lysis chamber 11 first, so that the detection sample can perform lysis reaction in the lysis chamber 11, after the lysis reaction is completed, the rotating member 40 can be rotated to communicate the liquid transfer chamber 21 with the lysis chamber 11, for example, a communication hole 401 communicating with the liquid transfer chamber 21 can be arranged on the rotating member 40, the rotation of the rotating member 40 can drive the communication hole 401 to rotate, when the communication hole 401 rotates to communicate with the first through hole 22, liquid in the lysis chamber 11 can flow into the communication hole 401 through the first through hole 22 and flow into the liquid transfer chamber 21 along the communication hole 401, so that liquid can be transferred from the lysis chamber 11 to the liquid transfer chamber 21.
S2, the liquid transferring cavity 21 obtains the liquid with the set quantity entering from the cracking cavity 11.
The pipetting cavity 21 is a space capable of containing liquid, and the volume of the space for containing liquid in the pipetting cavity 21 is constant and smaller than the volume of the lysis cavity 11, so that the space for containing liquid in the pipetting cavity 21 can be filled with liquid in the lysis cavity 11, that is, the liquid obtained by the pipetting cavity 21 is quantitative.
S3, the rotor 40 rotates to separate the pipetting cavity 21 from the lysis cavity 11 and to connect the pipetting cavity 21 with the amplification cavity 31.
After the liquid transferring cavity 21 obtains a fixed amount of liquid, the rotating member 40 is rotated to separate the liquid transferring cavity 21 from the lysis cavity 11, the lysis cavity 11 cannot transfer the liquid into the liquid transferring cavity 21, the rotating member 40 rotates to a certain position, the liquid transferring cavity 21 is communicated with the amplification cavity 31, and the liquid in the liquid transferring cavity 21 can enter the amplification cavity 31.
S4, the amplification chamber 31 receives a predetermined amount of liquid from the pipette chamber 21.
When the amplification chamber 31 is in communication with the pipetting chamber 21, the liquid in the pipetting chamber 21 can be transferred to the amplification chamber 31, and the volume of the amplification chamber 31 is at least equal to the volume of the pipetting chamber 21, so that the liquid in the pipetting chamber 21 can be completely transferred to the amplification chamber 31.
According to the embodiment of the invention, the rotating part rotates to control the communication and the separation among the liquid transferring cavity, the cracking cavity and the amplification cavity, so that liquid can be controlled to be transferred from the cracking cavity to the liquid transferring cavity and then from the liquid transferring cavity to the amplification cavity.
In some embodiments, before step S1, the method further includes: s0, adjusting the rotating member 40 to be at the initial position, wherein the initial position is the position for separating the pipetting cavity 21 from the lysis cavity 11 and the amplification cavity 31.
When the rotation member 40 is in the initial position, the pipetting chamber 21 may be closed so that both the lysis chamber 11 and the amplification chamber 31 are not communicated with the pipetting chamber 21 and thus the transfer of liquid is not performed, for example, when the rotation member 40 is in the initial position, the communication hole 401 is not communicated with either the first through hole 22 or the second through hole 23. The above description is only a preferred embodiment of the present invention, and is not intended to limit the scope of the present invention.
Claims (12)
1. A nucleic acid detecting apparatus, comprising:
a cracking piece, which is internally provided with a cracking cavity;
the pipetting piece is internally provided with a pipetting cavity, and the volume of the pipetting cavity is constant;
an amplification member having an amplification chamber therein;
a rotation member rotatable to close the pipetting chamber and open the pipetting chamber to communicate one of the lysis chamber and the amplification chamber.
2. The nucleic acid detecting apparatus according to claim 1, wherein the pipetting chamber communicates with the lysis chamber via a first channel, and the pipetting chamber communicates with the amplification chamber via a second channel.
3. The nucleic acid detecting device according to claim 2, wherein the rotating member includes a first rotating member and a second rotating member, the first rotating member is at least partially located in the lysis chamber and rotates to open and close the first channel, and the second rotating member is at least partially located in the amplification chamber and rotates to open and close the second channel.
4. The nucleic acid detecting device according to claim 3, wherein the first rotating member includes a first rotating disk, a first support member, and a second rotating disk arranged in this order in the first direction, and opposite ends of the first support member are connected to the first rotating disk and the second rotating disk, respectively; the second rotating disc is positioned in the cracking cavity, a first opening penetrating through the second rotating disc along the first direction is formed in the second rotating disc, and the first opening is communicated with or separated from the first channel along with the rotation of the first rotating part.
5. The nucleic acid detecting device according to claim 4, wherein an end of the first rotating disk remote from the second rotating disk protrudes from the lysis chamber.
6. The nucleic acid detecting apparatus according to claim 5, wherein the first rotating disk has a second opening extending therethrough in the first direction.
7. The nucleic acid detecting apparatus according to claim 3, wherein the second rotating member includes a third rotating disk, a second supporting member, and a fourth rotating disk arranged in this order in the first direction, and opposite ends of the second supporting member are connected to the third rotating disk and the fourth rotating disk, respectively; the third rotating disc is positioned in the amplification cavity, a third opening penetrating through the third rotating disc along the first direction is formed in the third rotating disc, and the third opening is communicated with or separated from the second channel along with the rotation of the second rotating element.
8. The apparatus for detecting nucleic acid according to claim 7, wherein an end of the fourth rotating disk facing away from the third rotating disk protrudes from the amplification chamber.
9. The nucleic acid detecting apparatus according to claim 1, wherein the lysis member, the pipetting member, and the amplification member are connected in this order in the first direction.
10. The nucleic acid detecting device according to any one of claims 1 to 9, wherein the first rotating member and the second rotating member are provided with sealing rings for sealing the lysis chamber and the amplification chamber, respectively.
11. A method for detecting a nucleic acid according to any one of claims 1 to 10, wherein the method for detecting comprises: s1, the rotating piece rotates to communicate the pipetting cavity with the lysis cavity and separate the pipetting cavity from the amplification cavity;
s2, the pipetting cavity acquires a set amount of liquid entering from the lysis cavity;
s3, the rotating piece rotates to separate the liquid transferring cavity from the cracking cavity and communicate the liquid transferring cavity with the amplification cavity;
s4, the amplification chamber obtains the set amount of liquid entering from the liquid transferring chamber.
12. The detecting method according to claim 11, before said step S1, further comprising:
and S0, adjusting the rotating piece to be at an initial position, wherein the initial position is a position at which the pipetting cavity is separated from the lysis cavity and the amplification cavity.
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| CN202111115363.6A CN113832014A (en) | 2021-09-23 | 2021-09-23 | Nucleic acid detection device and detection method |
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| CN202111115363.6A CN113832014A (en) | 2021-09-23 | 2021-09-23 | Nucleic acid detection device and detection method |
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