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EP-3940052-B1 - NUCLEIC ACID EXTRACTION APPARATUS

EP3940052B1EP 3940052 B1EP3940052 B1EP 3940052B1EP-3940052-B1

Inventors

  • CHEN, HUI

Dates

Publication Date
20260513
Application Date
20191119

Claims (18)

  1. A type of nucleic acid extraction device, comprising a workbench (1) with a tabletop (10), characterized by further comprising: a sliding seat (21) arranged on the tabletop (10) and capable of being driven by a sliding assembly (22) to slide along the tabletop (10) back and forth; a sample support (3) arranged on the sliding seat (21) and provided with a plurality of sample holes (31) formed side by side in a direction perpendicular to a sliding direction of the sliding seat (21), wherein the sample holes (31) are used for containing sample tubes (15); a reagent strip support (213) arranged on the sliding seat (21), wherein containing grooves (213a) for containing the reagent strips (7) are formed in the reagent strip support, the number of the containing grooves (213a) is consistent with that of the sample holes (3 1), each reagent strip (7) comprises the first sucker hole (71), the second sucker holes (72a and 72 b) and a reagent hole which are formed linearly, the first sucker holes (71) are used for containing sucker assemblies, the second sucker holes (72a and 72b) are used for containing suckers (82), the reagent holes comprise the first reagent holes (73) for containing diluent, the second reagent holes (74a, 74b, 74c and 74d) for containing cleaning fluid, the third reagent holes (75) for containing eluant and the fourth reagent holes (76) for storing target substances, and the first sucker holes (71), the second sucker holes (72a and 72b) and the reagent holes in the reagent strips (7) are opposite to the corresponding sample holes (31) correspondingly when the reagent strips (7) are put in the containing grooves (213a) correspondingly; a liquid transfer table (4) horizontally arranged on a vertical frame (12) fixed to the tabletop (10) and capable of being driven by a lifting assembly (64) to go up and down relative to the sliding seat (21); piston assemblies arranged on the liquid transfer table (4), wherein the piston assemblies comprise piston parts (62) and piston driving apparatuses (61), the number of the piston parts (62) is consistent with that of the sample holes (31), the piston parts (62) comprise piston tubes (622) and pistons (621) arranged in the piston tubes (622) and capable of moving along inner walls of the piston tubes (622) up and down, the piston tubes (622) all vertically penetrate through the liquid transfer table (4), the first connectors (621a) for being in inserted connection with the suckers (82) are formed at lower ends of the piston tubes (622) correspondingly, and the piston driving apparatuses (61) can drive the pistons (621) in the piston parts (62) to move up and down relative to the corresponding piston tubes (622); and suction assemblies comprising connecting suction tubes (91) and vacuum pumps (5), wherein the number of the connecting suction tubes (91) is consistent with that of the sample holes (31), the connecting suction tubes (91) all penetrate through the liquid transfer table (4), upper ends of the connecting suction tubes (91) connect with the vacuum pumps (5) correspondingly, and the second connectors (911) for being in inserted connection with the sucker assemblies are formed at lower ends of the connecting suction tubes correspondingly, wherein the sucker assemblies comprise filter element suckers (83) and disposable capillary suction tubes (81), adsorption filter elements (830) are embedded into the filter element suckers (83), upper ends of the disposable capillary suction tubes (81) sleeve lower ends of the connecting suction tubes (91) and are embedded into upper ports of the filter element suckers (83), and the disposable capillary suction tubes (81) are located in the filter element suckers (83) and located above the absorption filter elements (830).
  2. A nucleic acid extraction device according to claim 1, characterized in that a heating device for heating the sample support (3) is further arranged on the sliding seat.
  3. A nucleic acid extraction device according to claim 1 or 2, characterized by further comprising a puncture assembly for puncturing seals on the reagent holes, wherein the puncture assembly comprises puncture frames (141) and puncture heads (14) installed on the puncture frames (141), the puncture heads (14) match the sample holes (31), the pistons (621) are all installed on piston frames (63), the piston frames (63) are fixed to output shafts of the piston driving apparatuses (61), and the puncture frames (141) are fixed to the piston frames (63).
  4. A nucleic acid extraction device according to claim 3, characterized in that a plurality of vacuum pumps (5) correspond to piston parts (62) one to one, suction connectors (92) are arranged on the piston frames (63), the suction connectors (92) correspond to the piston parts (62) one to one in number, the suction connectors (92) all penetrate through the suction connector frames (65), one ends of suction connectors (92) are connected with connectors on the corresponding vacuum pumps (5) through connecting tubes, and the other ends of the suction connectors are connected with upper-end connectors of the connecting suction tubes (91) through connecting tubes.
  5. A nucleic acid extraction device according to claim 4, characterized in that an installation frame (51) is arranged on a tabletop (10), and the vacuum pumps (5) are put on the installation frame (51).
  6. A nucleic acid extraction device according to claim 1 or 2, characterized in that a sliding assembly (22) comprises the first sliding blocks (227), the first sliding rails (226), a pressing block (225), a sliding motor (221), a synchronous belt (224), a driving belt wheel (222) and a driven belt wheel (223); the first sliding blocks (227) are fixed to the bottom of the sliding seat (21), the first sliding rails (226) are fixed to the tabletop (10), the first sliding blocks (227) can slide along the first sliding rails (226) back and forth, the sliding motor (221) is fixed to the tabletop (10), the driving belt wheel (222) is fixed to an output shaft of the sliding motor (221), the driven belt wheel (223) is installed on the tabletop (10), the synchronous belt (224) is tensioned between the driving belt wheel (222) and the driven belt wheel (223), and the pressing block (225) is fixed to one side of the sliding seat (21) and pressed on the synchronous belt (224).
  7. A nucleic acid extraction device according to claim I or 2, characterized in that the lifting assembly (64) comprises a lifting motor (641), a lifting lead screw (642) and a lifting seat (643), the vertical frame (12) comprises an n-shaped frame (121) and a vertical plate (122) fixed to a top plate of the n-shaped frame (121), the lifting motor (641) is installed at the upper end of the vertical plate (122), the second sliding rail (122b) and a sliding hole (122a) vertically stretch from the position, below the lifting motor (641), of the vertical plate (122), the lifting lead screw (642) is vertically arranged, the upper end of the lifting lead screw is connected with an output shaft of the lifting motor (641), the lower end of the lifting lead screw is connected with the top plate of the n-shaped frame (121), the lifting seat (643) penetrates through the vertical plate (122) through the sliding hole (122a), the front end of the lifting seat is fixed to the liquid transfer table (4), the rear end of the lifting seat sleeves the lead screw (642), a screw transmission pair is formed by the lifting seat and the lead screw (642), the second sliding block (644) is arranged on the lifting seat (643), and when the lifting motor (641) drives the lead screw (642) to rotate, the lead screw (642) can drive the lifting seat (643), and enables the second sliding block (644) to slide along the second sliding rail (122b) up and down, and the corresponding position of the lifting seat (643) can move along the sliding hole (122a) up and down.
  8. A nucleic acid extraction device according to claim I or 2, characterized in that the reagent strips (7) comprise carrier plates (70), a plurality of the first through holes (701) and the second through holes (702) are formed side by side in the carrier plates (70) in a length direction of the carrier plates, tube bodies (703) corresponding to the first through holes (701) one to one stretch out of one sides of the carrier plates (70); meanwhile, sleeve bodies (704) corresponding to the second through holes (702) one to one further stretch out of one sides of the carrier plates (70), tube openings of the tube bodies (703) are right opposite to the corresponding first through holes (701) correspondingly, openings of the sleeve bodies (704) are right opposite to the corresponding second through holes (701) correspondingly, and therefore the first sucker holes (71), the second sucker holes (72a and 72b) and the reagent holes are formed correspondingly.
  9. A nucleic acid extraction device according to claim 8, characterized in that the first connecting parts (78) and the second connecting parts (77) are arranged on two sides of the carrier plates (70) correspondingly, and the first connecting parts (78) and the second connecting parts (77) can be detachably connected.
  10. A nucleic acid extraction device according to claim 9, characterized in that the first connecting parts (78) are connecting columns, and the second connecting parts (77) are connecting sleeves allowing the connecting columns to be inserted therein.
  11. A nucleic acid extraction device according to claim 1, characterized in that lower ends of the disposable capillary suction tubes (81) are opposite to the adsorption filter elements (830) up and down.
  12. A nucleic acid extraction device according to claim 11, characterized in that the distance between the lower ends of the disposable capillary suction tubes (81) and the upper surface of the adsorption filter elements (830) up and down is 0-10mm.
  13. A nucleic acid extraction device according to claim 12, characterized in that the distance between the lower ends of the disposable capillary suction tubes (81) and the upper surface of the adsorption filter elements (830) up and down is 2-5mm.
  14. A nucleic acid extraction device according to claim 1, characterized in that the disposable capillary suction tubes (81) comprise the first portions (811) located at the upper ends and the second portions (812) located at the lower ends, wherein the first portions (811) are used for being connected to the connecting suction tubes (91) in a sleeving mode, and tube diameters of the second portions (812) enable liquid to smoothly pass through the disposable capillary suction tubes (81) under the action of suction force.
  15. A nucleic acid extraction device according to claim 14, characterized in that tube diameters of tube bodies of the first portions (811) are 0.5-2.0mm.
  16. A nucleic acid extraction device according to claim 1, characterized in that edges of upper ports of the disposable capillary suction tubes (81) are lower than those of the upper ports of the filter element suckers (83).
  17. A nucleic acid extraction device according to claim 1, characterized in that convex rings (831) are arranged on inner circumferential surfaces of upper ends of the filter element suckers (83) in a circumferential direction, and the convex rings (831) abut against outer circumferential surfaces of corresponding positions of the disposable capillary suction tubes (81).
  18. A nucleic acid extraction device according to claim 17, characterized in that the number of the convex rings (831) arranged vertically is at least two, and the convex rings (831) are arranged vertically at intervals.

Description

Field of the Invention The invention relates to the technical field of biotechnologies, in particular to a type of nucleic acid extraction device. Background of the Invention Nucleic acid extraction is an important technology in life science research, biotechnology application and genetic diagnosis. Early nucleic acid extraction is established on the principle of phenol chloroform extraction for impurity removal and precipitation of nucleic acid with ethanol, but this method is very complex. At present, commercial nucleic acid extraction methods are mainly established on a solid phase adsorption method, in other words, nucleic acid molecules are adsorbed onto the surface of a certain solid phase adsorbent under the specific condition (e.g. pH and specific salt), and then elution purification methods are adopted, wherein two methods are mainly included: (1) Spin-column method: Spin-columns are filled with silicon-containing materials or other solids capable of adsorbing nucleic acid serving as media, guanidine salt and other chaotropic salt serve as binding agents; when passing through the spin-columns, the nucleic acid molecules are adsorbed onto the solid phases, then impurities are washed away through a solution containing ethanol, and finally nucleic acid is eluted out with a low-salt solution; and the method is simple and convenient, high in extraction efficiency and good in extraction effect, is a nucleic acid purification method most commonly used at present, but has the defects that repeated centrifugation is needed in the extraction process, and high-throughput and automatic operation is not convenient. (2) Magnetic bead method: Magnetic microspheres with surfaces wrapped with silicon materials or other materials capable of being bound with nucleic acid are used as solid phase carriers to separate DNA and RNA from samples under the action of chaotropic salt (guanidinium chloride, guanidinium isothiocyanate and the like) and external application of magnetic fields; and the method easily achieves automatic and high-throughput operation, but has the defects that the cost is high, time is wasted in the extraction process, thus, time consumed for whole genetic detection is too long, and doctors and patients cannot obtain detection results in time. The chemical stability of magnetic beads is poor. The extraction efficiency is easily affected, and the change of magnetic bead extraction effects of different samples, different manufacturers and different batches is large. As a result, extraction and purification results are not stable. In addition, cross-contamination is easily caused during repeated cleaning of the magnetic bead in the magnetic bead extraction process, leading to false positive results. Existing nucleic acid automatic extraction apparatuses are usually based on the magnetic bead method, such as nucleic acid extraction apparatuses disclosed in the China invention patent with the patent number of ZL2016020294747.7 (published application number of CN205635634U) and the China invention patent with the patent number of ZL201610687392.2 (published application number of CN106119082A). Japanese utility model JP3220438U discloses a sample extraction device having an extraction workbench and a movable operation module, wherein the adsorption process of nucleic acids on the adsorbent medium and the washing process of impurities on the adsorption membrane are accomplished by the movement of a syringe piston to draw in and press out liquid from a regular combination column. Content of the Invention The first technical problem to be solved by the invention is to provide a nucleic acid extraction device simple in structure and convenient to use for the prior art. The second technical problem to be solved by the invention is to provide a nucleic acid extraction device capable of extracting nucleic acid of high purity for the prior art. The third technical problem to be solved by the invention is to provide a nucleic 20 acid extraction device free of cross-contamination among samples in the nucleic acid extraction process for the prior art. The technical solution adopted by the invention to solve the above technical problem is: A type of nucleic acid extraction device comprises a workbench with a tabletop, and is characterized by further including: a sliding seat arranged on the tabletop and capable of being driven by a sliding assembly to slide along the tabletop back and forth;a sample support arranged on the sliding seat and provided with a plurality of sample holes formed side by side in a direction perpendicular to a sliding direction of the sliding seat, wherein the sample holes are used for containing sample tubes;a reagent strip support arranged on the sliding seat and provided with containing grooves for containing reagent strips, wherein the number of the containing grooves corresponds to that of the sample holes,each reagent strip comprising the first sucker holes, the second sucker holes and rea