CN115895879B - Nucleic acid extraction cartridge based on membrane adsorption method - Google Patents

Abstract

The application relates to the field of nucleic acid extraction instruments, in particular to a nucleic acid extraction cartridge based on a membrane adsorption method, which comprises an adsorption membrane, a support, a suction assembly and at least one reagent tube containing solution, wherein an adsorption cavity and at least one placing groove are arranged on the support, and a liquid channel is arranged between the adsorption cavity and the placing groove in a communicating manner; the adsorption film is arranged in the adsorption cavity, the reagent tube is placed in the placing groove, and the reagent tube can be communicated with the adsorption cavity through the liquid channel; the suction assembly is arranged on the support and is used for adjusting the pressure in the adsorption cavity so that the solution in the reagent tube can pass through the adsorption membrane. The utility model discloses a need not artifical injection solution many times and put into centrifuge with the reagent pipe in, can realize the process that the solution passes through the adsorption membrane to, the device can provide comparatively inclosed extraction environment for nucleic acid extraction, reduces the probability that the nucleic acid draws in-process polluted, reduces the probability that the virus gives off in the air simultaneously, improves the security when nucleic acid draws.

Description

Nucleic acid extraction cartridge based on membrane adsorption method

Technical Field

The invention relates to the field of nucleic acid extraction instruments, in particular to a nucleic acid extraction cartridge based on a membrane adsorption method.

Background

The method for extracting nucleic acid is a membrane adsorption method, the adsorption membrane is usually a silica gel membrane, the membrane adsorption method comprises the steps of cracking, washing, eluting and the like, when the method is operated, firstly, the collected sample is required to be mixed with a cracking solution to realize the separation of nucleic acid and protein, then the mixed solution passes through the adsorption membrane, the nucleic acid and part of impurities are adsorbed on the adsorption membrane, then the impurities on the adsorption membrane are washed off by washing liquid, and then the nucleic acid on the adsorption membrane is washed off by eluting liquid, so that a purified nucleic acid solution can be obtained and can be used for nucleic acid detection.

When the membrane adsorption method is used for nucleic acid detection, a solution pipe and an adsorption pipe are needed, an adsorption membrane is placed in the adsorption pipe, a small hole is formed in the bottom of the adsorption pipe, and the solution pipe is sleeved outside the adsorption pipe; during nucleic acid extraction, firstly injecting the cracked sample solution into an adsorption tube, then placing the adsorption tube into a centrifuge, and collecting the cracked solution in the solution tube through an adsorption film after centrifugation; at this time, the solution tube is replaced, and the washing solution and the eluting solution are sequentially injected into the adsorption tube, and the washing solution and the eluting solution are respectively passed through the adsorption membrane in the same manner until the purified nucleic acid solution is obtained. In the operation process, the solution needs to be injected into the adsorption tube for many times, the solution tube and the adsorption tube are placed into the centrifugal machine, the operation is troublesome, the adsorption membrane is always exposed in the air after adsorbing nucleic acid, the nucleic acid sample is easy to extract and pollute, and when the nucleic acid is extracted from some virus samples, viruses are easy to be dispersed into the air, so that a certain danger exists.

Disclosure of Invention

The purpose of this application is to provide a nucleic acid extraction card box based on membrane adsorption method, need not the manual work and pours into solution and put into centrifuge with the reagent pipe many times into, can realize the process that solution passes through the adsorption membrane into to, the device can provide comparatively inclosed extraction environment for nucleic acid extraction, reducible nucleic acid extraction in-process contaminated probability, reduces the probability that the virus gives off in the air simultaneously, improves the security when nucleic acid extraction.

The application provides a nucleic acid extraction cartridge based on membrane adsorption method adopts following technical scheme:

the nucleic acid extraction cartridge based on the membrane adsorption method comprises an adsorption membrane, a support, a suction assembly and at least one reagent tube for containing a solution, wherein an adsorption cavity and at least one placing groove are arranged on the support, and a liquid channel is arranged between the adsorption cavity and the placing groove in a communicating manner; the adsorption film is arranged in the adsorption cavity, the reagent tube is arranged in the placing groove, and the reagent tube can be communicated with the adsorption cavity through the liquid channel;

the suction assembly is arranged on the support and is used for adjusting the pressure in the adsorption cavity so that the solution in the reagent tube can pass through the adsorption membrane.

By adopting the technical scheme, the suction assembly works to form negative pressure in the adsorption cavity, so that the solution in the reagent tube enters the adsorption cavity through the liquid channel and passes through the adsorption membrane, and the suction assembly repeatedly sucks the solution in the reagent tube to repeatedly pass through the adsorption membrane and fully contact with the adsorption membrane; when the reagent tube filled with different solutions is placed in the placing groove, the different solutions can be contacted with the adsorption film, so that the steps of adsorbing, washing, eluting and the like of nucleic acid can be realized, the solution does not need to be manually injected or the reagent tube is manually placed in the centrifugal machine, and the operation is more convenient. And in addition, different solutions are directly stored in the reagent tube, manual injection is not needed, the tightness is good, the solution in the reagent tube enters the adsorption cavity through the liquid channel, the tightness is also good in the process, the probability of exposing viral nucleic acid to air can be reduced, the safety of the nucleic acid extraction process is improved, and meanwhile, the probability of being polluted during the nucleic acid sample extraction is reduced.

Optionally, the reagent tube sets up to three, is respectively for the first reagent tube of splendid attire lysate, be used for splendid attire lotion second reagent tube and be used for splendid attire eluent third reagent tube, the standing groove with reagent tube one-to-one sets up.

By adopting the technical scheme, the three reagent pipes respectively contain the lysate, the washing liquid and the eluent, and when one of the solutions is needed, the reagent pipes are communicated with the adsorption cavity, so that the operation is more convenient.

Optionally, a plurality of fixing holes are formed in the first side wall of the adsorption cavity, the plurality of fixing holes are distributed circumferentially, and the liquid channel is communicated with the fixing holes; the adsorption cavity is rotationally provided with a communication piece, the adsorption film is arranged on the communication piece, the communication piece is provided with a plurality of communication states, and the rotation of the communication piece can realize the switching of different communication states; the communication piece is provided with a communication hole, and when the communication piece is in the communication state, the communication hole is communicated with one of the fixing holes.

By adopting the technical scheme, when the communication hole is communicated with the fixed hole corresponding to the end part of the liquid channel, the solution in the corresponding reagent tube can enter the adsorption cavity through the liquid channel and pass through the adsorption film; the communicating piece rotates to enable the communicating hole to be communicated with the fixing holes at different positions, so that independent communication of different reagent pipes and the adsorption cavity can be realized, and the lysate, the washing liquid or the eluent can be respectively passed through the adsorption film, so that a plurality of steps of nucleic acid extraction are realized.

Optionally, a test tube is disposed on the support, and a collection channel is disposed between the test tube and one of the fixing holes.

By adopting the technical scheme, after the cracking, washing and eluting processes of nucleic acid extraction are completed, the obtained nucleic acid solution can be pumped into the test tube through the collecting channel, so that the subsequent nucleic acid test is more convenient.

Optionally, a mixing component for uniformly mixing the solution in the test tube with the freeze-drying reaction reagent is arranged on the test tube; the mixing component comprises a magnet and magnetic beads which are attracted by magnetism, the magnetic beads are positioned in the inspection tube, and the magnet is positioned outside the inspection tube.

By adopting the technical scheme, as the freeze-drying reaction reagent is required to be added in part of nucleic acid extraction, the freeze-drying reaction reagent can be placed in the test tube, and the purified nucleic acid solution is injected into the test tube to be mixed with the freeze-drying reaction reagent; the beads can be moved within the test tube by moving the magnet so that the mixing is more uniform.

Optionally, at least one of the fixing holes is communicated with the outside.

Through adopting above-mentioned technical scheme, when the inspection pipe is installed on the support, can exist a small amount of air in the inspection pipe, in the nucleic acid extraction process, will need carry out the evacuation to the inspection pipe before the eluent injection inspection pipe through the adsorption membrane, through setting up the fixed orifices with external intercommunication, can outwards discharge through this fixed orifices with the air that is taken out in the inspection pipe.

Optionally, a mounting groove is formed in the support and communicated with the adsorption cavity; the suction assembly comprises a cylinder body and a piston movably connected in the cylinder body, the cylinder body is arranged in the mounting groove, and the inner cavity of the cylinder body is communicated with the adsorption cavity; the cylinder is connected to the communicating member.

By adopting the technical scheme, the piston slides in the cylinder body to adjust the pressure of the adsorption cavity, so that the suction process of the solution is realized; the communicating piece is connected to the cylinder body, and the cylinder body can be operated to rotate outside the clamping box so as to drive the communicating piece to rotate, so that the communicating piece is controlled to rotate more conveniently.

Optionally, a positioning column is arranged on the support, a notch is arranged on the cylinder body, and when the notch is matched with the positioning column, the communicating piece is in the communicating state.

By adopting the technical scheme, the positioning column is clamped in the notch, so that the rotation of the cylinder body can be limited, and the processes of transportation, storage and the like of the cartridge can be facilitated; and the positioning column is clamped in the notch, the initial position of the communicating piece can be determined, and the initial position starts to rotate by a fixed angle, so that the communicating hole can be aligned with different fixing holes, and the conversion of different communicating states can be realized.

Optionally, a sealing film is disposed on a side of the reagent tube facing the placement groove, and a tip body is disposed in the placement groove.

By adopting the technical scheme, the reagent tube is inserted into the placing groove, and the sealing film can be pierced by the tip body, so that the solution in the reagent tube is released.

Optionally, a fence is arranged between the tip body and the side wall of the placing groove, a receiving bucket is arranged between the fence and the tip body, and a closed receiving cavity is formed between the receiving bucket and the reagent tube; the tip body is provided with an inlet channel, one end of the inlet channel is communicated with the liquid channel, and the other end of the inlet channel is communicated with the receiving cavity.

Through adopting above-mentioned technical scheme, tip body punctures the sealing membrane back, and solution in the reagent pipe can get into and accept the intracavity, and the inlet of tip body is submerged in solution, accepts chamber, entry passageway and absorption chamber all in airtight state this moment to when adjusting absorption chamber internal pressure, the solution in the controllable reagent pipe flows in or flows out the absorption chamber.

Drawings

FIG. 1 is a schematic diagram showing the overall structure of a nucleic acid extraction cartridge based on a membrane adsorption method according to an embodiment of the present application.

FIG. 2 is an exploded view of a nucleic acid extraction cartridge based on a membrane adsorption method according to an embodiment of the present application.

Fig. 3 is a schematic view of the internal structure of the support in the embodiment of the present application.

FIG. 4 is an overall schematic view of another angle of the membrane adsorption-based nucleic acid extraction cartridge according to the embodiment of the present application (to show the positional relationship between the reagent vessel and the pumping assembly).

FIG. 5 is a cross-sectional view taken along A-A in FIG. 4 (wherein the first reagent vessel is in a state of being inserted into the placement groove).

Reference numerals illustrate:

1. a support; 11. a placement groove; 12. an adsorption chamber; 13. a liquid channel; 14. a base; 141. a reagent section; 142. a suction unit; 15. a bottom plate; 151. an exhaust hole; 16. a mounting groove; 17. a fixing hole; 171. a first fixing hole; 172. a second fixing hole; 173. a third fixing hole; 174. a fourth fixing hole; 175. a fifth fixing hole; 18. a collection channel; 2. a reagent tube; 21. a first reagent tube; 211. sealing film; 212. a stabilizing sleeve; 22. a second reagent tube; 23. a third reagent tube; 3. an adsorption film; 4. a suction assembly; 41. a cylinder; 42. a piston; 43. a piston rod; 5. a tip body; 51. a surrounding baffle; 511. an embedding groove; 52. a receiving bucket; 521. a receiving cavity; 53. an inlet passage; 54. a liquid inlet; 6. a communication member; 61. a groove; 62. a communication hole; 7. a positioning plate; 71. a notch; 72. positioning columns; 8. a test tube; 81. a thread sleeve; 9. mixing the components uniformly; 91. magnetic beads; 92. and (3) a magnet.

Detailed Description

The present application is described in further detail below in conjunction with figures 1-5.

The embodiment of the application discloses a nucleic acid extraction cartridge based on a membrane adsorption method. Referring to fig. 1 to 3, a nucleic acid extraction cartridge based on a membrane adsorption method includes a support 1, a reagent tube 2, an adsorption membrane 3, and a suction assembly 4, a placement groove 11 and an adsorption chamber 12 are provided on the support 1, the reagent tube 2 is placed in the placement groove 11, the adsorption membrane 3 is provided in the adsorption chamber 12, a liquid channel 13 is provided between the placement groove 11 and the adsorption chamber 12, and the placement groove 11 can be communicated with the adsorption chamber 12 through the liquid channel 13. The suction component 4 is arranged on the support 1, and when the suction component 4 works, the pressure in the adsorption cavity 12 can be adjusted, so that the solution in the reagent tube 2 enters the adsorption cavity 12 through the liquid channel 13 and passes through the adsorption membrane 3, thereby realizing the processes of adsorbing, washing, eluting and the like of nucleic acid, and being more convenient to operate. In addition, the solution in the reagent tube 2 enters the adsorption cavity 12 through the liquid channel 13 to be contacted with the adsorption film 3, so that the probability of exposing the solution in the external environment can be reduced, and the possibility of being polluted during the extraction of the nucleic acid sample is reduced.

Referring to fig. 1-3, in this embodiment, three reagent tubes 2 are provided, three placement grooves 11 are provided on the support 1, and the placement grooves 11 are in one-to-one correspondence with the reagent tubes 2; the three liquid channels 13 are also arranged, the three placing grooves 11 are respectively communicated with the adsorption cavity 12 through the corresponding liquid channels 13, and the three liquid channels 13 are arranged in a staggered manner and are not communicated with each other, so that the liquid in the three reagent tubes 2 can flow into the adsorption cavity 12 through the corresponding liquid channels 13 and pass through the adsorption membrane 3. The three reagent tubes 2 are respectively a first reagent tube 21, a second reagent tube 22 and a third reagent tube 23, wherein the first reagent tube 21 is used for containing a lysate, the volume of the lysate is 200 mu L, the second reagent tube 22 is used for containing a washing solution, the volume of the washing solution is 400 mu L, the third reagent tube 23 is used for containing an eluent, and the volume of the eluent is 200 mu L; at the beginning of nucleic acid extraction, it is necessary to inject a sample into the first reagent tube 21, and then sequentially aspirate the solutions in the first reagent tube 21, the second reagent tube 22, and the third reagent tube 23 into the adsorption chamber 12. In other application scenarios, the reagent tube 2 may be set to other numbers depending on the number of reagent species required for nucleic acid extraction.

Referring to fig. 1, 3 and 4, the support 1 is made of plastic material, the support 1 includes a base 14 and a bottom plate 15, the base 14 includes an integrally formed reagent portion 141 and a suction portion 142, and three placement grooves 11 are sequentially arranged in parallel in the reagent portion 141; the suction part 142 is provided with a mounting groove 16, the length of the mounting groove 16 along the axial direction of the suction part is greater than that of the placing groove 11 along the axial direction of the suction part, the suction assembly 4 is mounted in the mounting groove 16, the placing groove 11 and the mounting groove 16 are cylindrical, the axes of the three placing grooves 11 are coplanar, the axis of the mounting groove 16 is parallel to the plane where the axes of the three placing grooves 11 are located, and the third reagent tube 23, the second reagent tube 22 and the first reagent tube 21 are sequentially placed in the three placing grooves 11 along the direction away from the mounting groove 16.

Referring to fig. 1 and 2, a side surface of the suction part 142 perpendicular to the axis of the mounting groove 16 is flush with a side surface of the reagent part 141 perpendicular to the placement groove 11, the side surface of the suction part 142 flush with the reagent part 141 is a bottom surface of the base 14, the liquid channel 13 is disposed on the bottom surface of the base 14, and the bottom plate 15 is attached to and can be fixedly connected to the bottom surface of the base 14 by gluing, screwing or the like, so that the liquid channel 13 is a closed cavity, and the support 1 can be placed on a horizontal surface through the bottom plate 15.

Referring to fig. 4 and 5, the second reagent tube 22 has a cylindrical shape, one end of which is closed, and the other end of which is covered with a sealing film 211, so that a closed chamber is formed inside the second reagent tube 22; one end of the second reagent tube 22 in a closed state is provided with a force application part which is cross-shaped so as to be convenient to take; the end of the second reagent tube 22 covered with the sealing film 211 is used for releasing the washing liquid into the placing groove 11, and when the second reagent tube 22 is installed, the end provided with the sealing film 211 is required to face the placing groove 11; the end of the second reagent vessel 22 near the placement groove 11 is integrally provided with a stabilizing sleeve 212 for cooperation with the placement groove 11, and the inner diameter of the stabilizing sleeve 212 is larger than the outer diameter of the first reagent vessel 21.

Referring to fig. 4 and 5, the third reagent vessel 23 has the same shape, structure, and size as the second reagent vessel 22.

Referring to fig. 1, 4 and 5, the first reagent vessel 21 is similar in structure to the second reagent vessel 22, except for: the end cover of the first reagent tube 21 far away from the placing groove 11 is provided with a cover body, and the cover body is removed, so that the collected nucleic acid sample liquid can be injected into the first reagent tube 21 to realize the nucleic acid cracking. In addition, two parts are cut relatively on the outer circular side wall of the stabilizing sleeve 212 connected to the first reagent tube 21 to form two planes, the inner side wall of the placing groove 11 far away from the mounting groove 16 (i.e. the placing groove 11 for inserting the first reagent tube 21) is also provided with two planes, when the first reagent tube 21 is inserted into the placing groove 11, the planes on the stabilizing sleeve 212 are attached to the planes on the placing groove 11, so that the rotation of the stabilizing sleeve 212 can be limited, the position of the first reagent tube 21 is stable, and meanwhile, a inspector can distinguish the reagent tube 2 as the first reagent tube 21, and the sample liquid needs to be injected into the reagent tube 2.

Referring to fig. 3 to 5, when the reagent vessel 2 is inserted into the placement groove 11, in order to facilitate piercing of the sealing film 211 on the reagent vessel 2 so that the solution enters the liquid passage 13, the placement groove 11 is provided through the base 14, and the tip body 5 is provided in the placement groove 11, which will be described in detail below with reference to the first reagent vessel 21 as an example. The tip body 5 is cylindrical and is coaxially arranged with the placing groove 11, the end face of the tip body 5 close to the bottom surface of the seat body 14 is flush with the bottom surface of the seat body 14, and the end face of the tip body far away from the bottom surface of the seat body 14 is conical and faces the sealing film 211; a fence 51 is further arranged between the tip end body 5 and the side wall of the placing groove 11, the fence 51 is coaxially sleeved outside the tip end, one end face of the fence 51 is flush with the bottom face of the seat body 14, and the fence and the seat body 14 are integrally formed; an embedded groove 511 is formed between the enclosure 51 and the side wall of the placement groove 11, and when the first reagent tube 21 is inserted into the placement groove 11, the stabilizing sleeve 212 can be inserted into the embedded groove 511, so that the position of the first reagent tube 21 in the placement groove 11 is more stable.

Referring to fig. 3 and 5, one end of the enclosure 51 near the first reagent tube 21 is flared in a cone shape so that one end of the enclosure 51 near the first reagent tube 21 can be abutted against the inner wall of the stabilizing sleeve 212, the first reagent tube 21 is placed in the placing groove 11, the stabilizing sleeve 212 is clamped between the enclosure 51 and the side wall of the placing groove 11, and the first reagent tube 21 can be inserted into the placing groove 11 by means of external force; and, an end of the inner wall of the stabilizing jacket 212 remote from the first reagent tube 21 is provided with a slope so that the stabilizing jacket 212 is inserted into the insertion groove 511.

Referring to fig. 4 and 5, the enclosure 51 is arranged in a clearance with the tip body 5, a receiving hopper 52 is arranged between the enclosure 51 and the tip body 5, the receiving hopper 52 is conical and is arranged with an opening facing the tip of the tip body 5, and the enclosure 51, the receiving hopper 52 and the tip body 5 are integrally formed; when the first reagent vessel 21 is completely inserted into the placement groove 11, the end of the enclosure 51 away from the bottom surface of the housing 14 is in close contact with the end surface of the first reagent vessel 21 where the sealing film 211 is provided, and at this time, the tip body 5 can pierce the sealing film 211 and be inserted into the first reagent vessel 21.

Referring to fig. 2, 3 and 5, the end surfaces of the enclosure 51, the receiving hopper 52 and the first reagent tube 21 on which the sealing film 211 is provided form a closed receiving chamber 521, and the solution in the first reagent tube 21 can flow into the receiving chamber 521. An inlet channel 53 is arranged in the tip body 5, one end of the inlet channel 53 extends to the bottom surface of the base body 14 and is communicated with the liquid channel 13, and one end of the inlet channel 53 away from the liquid channel 13 extends to the receiving cavity 521; the part of the tip body 5 in the receiving cavity 521 is provided with four liquid inlets 54, the four liquid inlets 54 are uniformly distributed along the circumferential direction of the tip body 5 and are communicated with the inlet channel 53, and the solution in the receiving cavity 521 and the first reagent tube 21 can enter the inlet channel 53 through the liquid inlets 54 and further enter the liquid channel 13.

Referring to fig. 4 and 5, the suction assembly 4 is for sucking the solution in the reagent vessel 2 into the adsorption chamber 12, and the suction assembly 4 includes a cylinder 41, a piston 42, and a piston rod 43; the cylinder 41 is penetrated in the mounting groove 16, the piston rod 43 is penetrated in the cylinder 41 in a sliding way, and the piston 42 is fixedly connected with the end part of the piston rod 43; the piston 42 can be made of rubber material, one end of the piston 42 close to the bottom surface of the base 14 is provided with a cone shape, and the inner wall of the cylinder 41 is also provided with a cone shape, so that the piston 42 can be tightly attached to the inner wall of the cylinder 41; the end of the piston rod 43 far away from the piston 42 extends out of the cylinder 41, and the pressure of the cavity inside the cylinder 41 can be adjusted by pulling the piston rod 43.

Referring to fig. 4 and 5, the adsorption cavity 12 is located at a position of the mounting groove 16 near the bottom surface of the base 14 and is disposed in communication with the mounting groove 16, and specifically, the adsorption cavity 12 may be integrally formed with the mounting groove 16; the end wall of the cylinder 41 near the bottom surface of the seat 14 is provided with a through hole, and the inner cavity of the cylinder 41 is communicated with the adsorption cavity 12 through the through hole, so that the pressure in the adsorption cavity 12 can be regulated by pulling the piston rod 43.

Referring to fig. 2 and 3, in order to enable the adsorption chamber 12 to be respectively communicated with different reagent tubes 2, five fixing holes 17 are formed in a first side wall of the adsorption chamber 12, the five fixing holes 17 are uniformly distributed circumferentially about an axis of the mounting groove 16, and are sequentially named as a first fixing hole 171, a second fixing hole 172, a third fixing hole 173, a fourth fixing hole 174 and a fifth fixing hole 175 in an arrangement order, a liquid channel 13 corresponding to the first reagent tube 21 is communicated with the first fixing hole 171, a liquid channel 13 corresponding to the second reagent tube 22 is communicated with the third fixing hole 173, and a liquid channel 13 corresponding to the third reagent tube 23 is communicated with the second fixing hole 172.

Referring to fig. 2 and 3, the adsorption chamber 12 is provided with a communicating member 6, the communicating member 6 is cylindrical and has a cylindrical groove 61 formed therein, and the adsorption film 3 is cylindrical and is embedded in the groove 61 of the communicating member 6; the communicating piece 6, the adsorption cavity 12 and the mounting groove 16 are coaxial, and the communicating piece 6 is rotatably mounted around the axis of the mounting groove 16; the side wall of the communicating piece 6 close to the bottom plate 15 is in a closed state, the closed side wall is the bottom wall of the communicating piece 6, the bottom wall of the communicating piece 6 is provided with a communicating hole 62, and the diameter of the communicating hole 62 is equal to that of the fixing hole 17; the axis of the communication hole 62 and the axes of the five fixing holes 17 are located on the same cylindrical surface, the cylindrical surface is centered on the axis of the mounting groove 16, the communication member 6 is rotated, and the communication hole 62 can communicate with the fixing holes 17 at different positions.

Referring to fig. 2 and 3, when the communication hole 62 communicates with the fixed hole 17, the communication piece 6 is in a communication state, and the position of the communication piece 6 in the communication state is a communication position, and the communication positions are named as a first communication position, a second communication position, a third communication position, a fourth communication position, and a fifth communication position according to the naming order of the fixed hole 17; when the communicating member 6 is in the first communicating position, the communicating hole 62 is aligned with the first fixing hole 171, the liquid in the first reagent tube 21 can enter the groove 61 of the communicating member 6 through the liquid channel 13, when the communicating member 6 is in the second communicating position, the communicating hole 62 is aligned with the second fixing hole 172, and so on, the communicating member 6 can realize the conversion of the adjacent communicating position every 72 degrees of rotation; the communication member 6 rotates forward from the first communication position to the second communication position, and rotates backward from the second communication position to the first communication position.

Referring to fig. 4 and 5, in order to facilitate the control of the rotation of the communicating element 6, a cylindrical sleeve integrally extends from the end surface of the cylinder 41 near the bottom plate 15, the outer diameter of the cylindrical sleeve is smaller than that of the cylinder 41, the adsorption film 3 is embedded in the cylindrical sleeve, the communicating element 6 can be made of rubber material and sleeved outside the cylindrical sleeve, and the rotating cylinder 41 can drive the communicating element 6 and the adsorption film 3 to rotate. Referring to fig. 1, in order to determine the position of the communicating element 6, a positioning disc 7 is fixedly sleeved on the cylinder 41, a circular notch 71 is arranged on the positioning disc 7, a positioning column 72 is fixedly arranged on the end surface of the suction part 142 far away from the bottom plate 15, and when the positioning column 72 is penetrated in the notch 71, the communicating element 6 is in a first communicating position; the first communicating position is the initial position of the communicating piece 6, and the communicating piece 6 rotates by a fixed angle from the initial position, so that the conversion of different communicating positions can be realized. The positioning disk 7 is provided with a fluted disk which can be connected with other equipment so as to control the rotation of the cylinder 41.

Referring to fig. 1 and 2, in order to facilitate the detection of purified nucleic acid, a detection tube 8 is provided on the side of the bottom plate 15 remote from the base 14, the portion of the detection tube 8 close to the bottom plate 15 is cylindrical, the portion remote from the bottom plate 15 is conical and the end remote from the bottom plate 15 is spherical; a threaded sleeve 81 is integrally arranged on the bottom surface of the bottom plate 15, which is far away from the base 14, and a threaded hole of the threaded sleeve 81 penetrates through the bottom plate 15; the part of the inspection tube 8 close to the support 1 is provided with external threads, and the inspection tube 8 can be threaded in the threaded sleeve 81 so as to realize the disassembly and assembly of the inspection tube 8.

Referring to fig. 2 and 3, the bottom surface of the base 14 is provided with a collecting channel 18, the collecting channel 18 extends from the fifth fixing hole 175 to the position of the inspection tube 8, the end surface of the inspection tube 8 near the end surface provided with the external thread can be abutted against the bottom surface of the base 14, at this time, the end of the collecting channel 18 is aligned with the center of the inspection tube 8, and the solution in the adsorption cavity 12 can enter the inspection tube 8 through the collecting channel 18.

Referring to FIGS. 2 and 3, for some nucleic acid extractions it is necessary to add a lyophilization reagent, such as a protease, to the purified nucleic acid to aid in detection, in which case it is necessary to add the lyophilization reagent to the assay tube 8. In order to facilitate the easy mixing of purified nucleic acid and freeze-dried reaction reagent, a mixing component 9 is arranged on the test tube 8; the mixing component 9 comprises magnetic beads 91 and a magnet 92, wherein the magnetic beads 91 are attracted by magnetism, the magnetic beads 91 are placed in the inspection tube 8, the magnet 92 is placed outside the inspection tube 8, the magnet 92 is attracted by the magnetic beads 91, the magnet 92 moves back and forth along the axial direction of the inspection tube 8, and the magnetic beads 91 can move back and forth in the inspection tube 8 along with the axial direction of the inspection tube 8 so as to achieve the mixing effect; the diameter of the magnetic beads 91 is not larger than the spherical inner diameter of the end of the test tube 8 remote from the bottom plate 15 so that the magnetic beads 91 can reach the bottom of the test tube 8.

In other application scenarios, if nucleic acid extraction does not require the addition of lyophilization reagents, the homogenization assembly 9 may not be provided.

Referring to fig. 2 and 3, since the test tube 8 has air inside at the time of installation, before the eluent is injected into the test tube 8, it is necessary to vacuumize the test tube 8 and discharge the extracted air through the fourth fixing hole 174; specifically, the bottom plate 15 is provided with the exhaust hole 151, the exhaust hole 151 is aligned with the fourth fixing hole 174, and when the communication hole 62 is in the fourth communication position, the communication hole 62 is aligned with the fourth fixing hole 174, so that the adsorption chamber 12 can communicate with the external environment. In other application scenarios, if nucleic acid extraction is performed on a viral sample, the fourth fixing hole 174 may not be provided, and the air in the extracted test tube 8 may be injected into the first reagent tube 21 or the second reagent tube 22 to reduce the possibility of virus entering the air.

The nucleic acid extraction cartridge based on the membrane adsorption method of this embodiment is used in combination with a nucleic acid detecting instrument, so that except for the fact that the injection of the sample liquid in the first reagent tube 21 is manually completed, the three reagent tubes 2 are pushed down and pierced through the sealing membrane 211, the drawing of the piston rod 43, the rotation of the cylinder 41 and the sliding process of the magnet 92 are controlled by the nucleic acid detecting instrument to act more precisely. In other application scenarios, the nucleic acid extraction cartridge based on the membrane adsorption method may be used alone, and the reagent tube 2 may be pushed down and pierced through the sealing membrane 211, the drawing of the piston rod 43, the rotation of the cylinder 41, and the sliding of the magnet 92 may be manually operated.

The initial state of a nucleic acid extraction cartridge based on a membrane adsorption method according to the embodiment of the application is: the three reagent pipes 2 are all placed in the placing groove 11, the positioning column 72 is embedded in the notch 71, the communicating piece 6 is in a first communicating position, the placing groove 11 below the first reagent pipe 21 is communicated with the adsorption cavity 12, and the method mainly comprises the following steps in actual operation:

the first step: 200. Mu.L of the sample solution was injected into the first reagent tube 21 and sealed with a cap;

and a second step of: simultaneously, the first reagent tube 21, the second reagent tube 22 and the third reagent tube 23 are pressed down, the stabilizing sleeve 212 is inserted into the embedding groove 511, the sealing films 211 on the three reagent tubes 2 are simultaneously pierced, and the three reagent tubes 2 are respectively communicated with the corresponding liquid channels 13;

and a third step of: the piston rod 43 is pulled a plurality of times, the number of times of pulling can be set to 8-15 times, and 3 seconds are required each time (the lysate is fully mixed with the sample solution and passes through the adsorption film 3 a plurality of times, and the cracked nucleic acid and part of impurities are adsorbed on the adsorption film 3);

fourth step: when the piston rod 43 is pressed down to the limit position (the pyrolysis liquid flows back to the first reagent tube 21), the cylinder 41 rotates forward for 72 degrees twice (the positioning column 72 is broken), and the communicating member 6 is converted from the first communicating position to the third communicating position;

fifth step: repeating the third step (the washing liquid passes through the adsorption film 3 for a plurality of times, so that impurities on the adsorption film 3 can be washed away);

sixth step: when the piston rod 43 is depressed to the limit position (the washing liquid flows back to the first reagent tube 21), the cylinder 41 is rotated forward by 72 ° twice, and the communicating element 6 is switched from the third communicating position to the fifth communicating position;

seventh step: pulling the piston rod 43 outwards and evacuating the test tube 8;

eighth step: the cylinder 41 reversely rotates for 72 degrees, the communicating piece 6 is converted from a fifth communicating position to a fourth communicating position, the piston rod 43 is pressed down to the limit position in the cylinder 41, and air is discharged;

ninth step: the cylinder 41 rotates forward for 72 degrees twice, and the communicating piece 6 is converted into a second communicating position from a fourth communicating position;

tenth step: repeating the third step (the eluent passes through the adsorption membrane 3 a plurality of times, so that the nucleic acid on the adsorption membrane 3 can be washed out and dissolved in the eluent);

eleventh step: sucking the eluent into the adsorption cavity 12 and the cylinder 41, reversely rotating the cylinder 41 for 72 degrees for three times, converting the communication piece 6 from the second communication position to the fifth communication position, and then pumping the eluent into the inspection tube 8;

twelfth step: the magnet 92 is reciprocally slid, and the nucleic acid solution and the lyophilized reaction reagent are uniformly mixed by the magnetic beads 91, and then the nucleic acid solution in the test tube 8 can be tested.

The foregoing are all preferred embodiments of the present application, and are not intended to limit the scope of the present application in any way, therefore: all equivalent changes in structure, shape and principle of this application should be covered in the protection scope of this application.

Claims (7)

1. A nucleic acid extraction cartridge based on a membrane adsorption method, comprising an adsorption membrane (3), characterized in that: the device comprises a support (1), a suction assembly (4) and at least one reagent tube (2) for containing a solution, wherein an adsorption cavity (12) and at least one placing groove (11) are arranged on the support (1), a liquid channel (13) is communicated between the adsorption cavity (12) and the placing groove (11), and the liquid channel (13) is a closed cavity and is arranged on the support (1); the adsorption film (3) is arranged in the adsorption cavity (12), the reagent tube (2) is arranged in the placing groove (11), and the reagent tube (2) can be communicated with the adsorption cavity (12) through the liquid channel (13);

the suction assembly (4) is arranged on the support (1), and the suction assembly (4) is used for adjusting the pressure in the adsorption cavity (12) so that the solution in the reagent tube (2) can pass through the adsorption membrane (3);

a plurality of fixing holes (17) are formed in the first side wall of the adsorption cavity (12), the plurality of fixing holes (17) are distributed circumferentially, and the liquid channel (13) is communicated with the fixing holes (17);

the adsorption cavity (12) is rotationally provided with a communication piece (6), a groove (61) is formed in the communication piece (6), the suction assembly (4) comprises a cylinder body (41) and a piston (42), the piston (42) is movably installed in the cylinder body (41), a cylinder sleeve is arranged on the end face, close to the communication piece (6), of the cylinder body (41), the adsorption film (3) is embedded in the cylinder sleeve, and the communication piece (6) is sleeved outside the cylinder sleeve through the groove (61);

the communicating piece (6) is provided with a plurality of communicating states, and the rotation of the communicating piece (6) can realize the switching of different communicating states; a communication hole (62) is formed in the communication piece (6), and when the communication piece (6) is in the communication state, the communication hole (62) is communicated with one of the fixing holes (17);

a sealing film (211) is arranged on one side, facing the placing groove (11), of the reagent tube (2), a tip body (5) is arranged in the placing groove (11), and a stabilizing sleeve (212) is arranged at one end, close to the tip body (5), of the reagent tube (2);

a fence (51) is arranged between the tip body (5) and the side wall of the placing groove (11), the stabilizing sleeve (212) is sleeved outside the fence (51), and one end, close to the reagent tube (2), of the fence (51) is abutted against the inner wall of the stabilizing sleeve (212); a receiving bucket (52) is arranged between the enclosing block (51) and the tip end body (5), and a closed receiving cavity (521) is formed between the receiving bucket (52) and the reagent tube (2); an inlet channel (53) is arranged on the tip body (5), one end of the inlet channel (53) is communicated with the liquid channel (13), and the other end of the inlet channel is communicated with the receiving cavity (521).

2. The membrane adsorption-based nucleic acid extraction cartridge of claim 1, wherein: the reagent tubes (2) are three, namely a first reagent tube (21) for containing lysate, a second reagent tube (22) for containing washing liquid and a third reagent tube (23) for containing eluent, and the placing grooves (11) are arranged in one-to-one correspondence with the reagent tubes (2).

3. The membrane adsorption-based nucleic acid extraction cartridge of claim 1, wherein: the support (1) is provided with a checking tube (8), and a collecting channel (18) is communicated between the checking tube (8) and one of the fixing holes (17).

4. The membrane adsorption-based nucleic acid extraction cartridge according to claim 3, wherein: a freeze-drying reaction reagent is arranged in the test tube (8), and a mixing component (9) for uniformly mixing the solution in the test tube (8) with the freeze-drying reaction reagent is arranged on the test tube (8); the mixing component (9) comprises a magnet (92) and magnetic beads (91) which are attracted by magnetism, the magnetic beads (91) are located in the inspection tube (8), and the magnet (92) is located outside the inspection tube (8).

5. The membrane adsorption-based nucleic acid extraction cartridge of claim 4, wherein: at least one of the fixing holes (17) is communicated with the outside.

6. The membrane adsorption-based nucleic acid extraction cartridge of claim 1, wherein: the support (1) is communicated with the adsorption cavity (12) and provided with a mounting groove (16); the cylinder (41) is mounted to the mounting groove (16).

7. The membrane adsorption-based nucleic acid extraction cartridge of claim 6, wherein: the support (1) is provided with a positioning column (72), the cylinder (41) is provided with a notch (71), and when the notch (71) is matched with the positioning column (72), the communicating piece (6) is in the communicating state.

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