WO2016112694A1 - 一种磁珠法核酸提取装置 - Google Patents

一种磁珠法核酸提取装置 Download PDF

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WO2016112694A1
WO2016112694A1 PCT/CN2015/087791 CN2015087791W WO2016112694A1 WO 2016112694 A1 WO2016112694 A1 WO 2016112694A1 CN 2015087791 W CN2015087791 W CN 2015087791W WO 2016112694 A1 WO2016112694 A1 WO 2016112694A1
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magnet
nucleic acid
acid extraction
disposed
sample
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PCT/CN2015/087791
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English (en)
French (fr)
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章洪建
戴立忠
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湖南圣维基因科技有限公司
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Priority to EP15877607.0A priority Critical patent/EP3222710B1/en
Publication of WO2016112694A1 publication Critical patent/WO2016112694A1/zh
Priority to US15/622,673 priority patent/US9896684B2/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
    • C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
    • C12N15/1013—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by using magnetic beads
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
    • C12M47/06—Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C1/00—Magnetic separation
    • B03C1/02—Magnetic separation acting directly on the substance being separated
    • B03C1/04—Magnetic separation acting directly on the substance being separated with the material carriers in the form of trays or with tables
    • B03C1/06—Magnetic separation acting directly on the substance being separated with the material carriers in the form of trays or with tables with magnets moving during operation
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C2201/00—Details of magnetic or electrostatic separation
    • B03C2201/18—Magnetic separation whereby the particles are suspended in a liquid

Definitions

  • the invention relates to the field of nucleic acid extraction devices, in particular to a magnetic bead nucleic acid extraction device
  • molecular level detection can significantly shorten the detection window and improve detection sensitivity.
  • the quality of the analyte obtained from the specimen is an important factor affecting the test results.
  • the amount and purity of the nucleic acid extracted from the specimen directly affects the results of subsequent PCR (polymerase chain reaction) detection.
  • the extraction of nucleic acids generally involves the following steps: 1) cleavage of the membrane of the coated nucleic acid and release of the nucleic acid; 2) removal of the protein; 3) collection of the nucleic acid.
  • the classic methods are boiling lysis and phenol-chloroform extraction. The boiling lysis method firstly adds a precipitant to the sample, and then discards the supernatant to remove the small molecule inhibitor and precipitate the virus particles; then, the lysate is added and boiled to release the macromolecular inhibitor such as DNA and precipitated residual protein, and centrifuged. Clear is the DNA obtained.
  • the phenol-chloroform extraction method firstly adds the lysate to the sample, then adds chloroform and then centrifuges to release the RNA and separate it from the protein layer, and then the supernatant is added to the isopropanol to extract the RNA, and the supernatant is discarded after centrifugation.
  • the obtained RNA is added after washing with ethanol.
  • a nano-sized particle (magnetic bead) is used in the nucleic acid extraction process.
  • This kind Paramagnetic nanoparticles can specifically or non-specifically adsorb nucleic acids, and magnetic fields can be used to conveniently separate nucleic acids from the aqueous phase for the purpose of extracting nucleic acids.
  • the step of nucleic acid extraction specifically comprises: in the nucleic acid-containing sample, 1) lysis: adding the lysate, the sample is cleaved out of the nucleic acid, 2) binding: adding the magnetic beads, the magnetic beads specifically bind the nucleic acid, 3) washing: adding The washing liquid removes impurities such as proteins on the nucleic acid by a washing step, 4) elution: separating the magnetic beads and the nucleic acid by adding an eluent, and 5) separating: separating the magnetic beads under a magnetic field to obtain a purified nucleic acid.
  • lysis adding the lysate
  • binding adding the magnetic beads
  • the magnetic beads specifically bind the nucleic acid
  • washing liquid removes impurities such as proteins on the nucleic acid by a washing step
  • elution separating the magnetic beads and the nucleic acid by adding an eluent
  • a lysate, a magnetic bead, and a sample are placed in a beaker (or a sample tube), and a magnetic bar (rod magnet) and a disposable outer sleeve (Tip) placed on one end of the magnetic bar are used together to place a beaker containing the above solution.
  • the magnetic rod and its outer sleeve are slowly moved up and down to collect a combination of the magnetic beads and the nucleic acid, and then the magnetic rod and the outer sleeve are transferred into another solution (washing liquid), and the magnetic rod is first taken out, and the magnetic field disappears.
  • the jacket moves up and down rapidly in the wash solution, and the combination of magnetic beads and nucleic acids enters the solution.
  • the magnetic rod is then jacketed to collect the combination of magnetic beads and nucleic acids, and the magnetic rod and the outer casing are separated in the eluent. Magnetic beads were collected in such a manner that the solution containing the magnetic beads and the nucleic acid was returned to the beaker to obtain a pure nucleic acid solution.
  • the combination of a magnet and a disposable jacket can complete the transfer of magnetic beads from one reagent to another.
  • the above rod magnets are mainly arrayed (2 columns * 8 rows, 4 columns * 8 rows or 12 columns * 8 rows), and the array pattern of the disposable jacket is 1 * 8 or 12 * 8 .
  • the reaction tube was a 96-well deep well plate. From the way of working, it can be divided into two types: one, using a 2*8 or 4*8 magnetic rod array and a 1*8 disposable jacket to complete the extraction process of 16 or 32 samples in a deep well plate. . Second, using a 12*8 magnetic rod array and a 12*8 disposable jacket to complete an extraction step (a reagent) in a deep well plate, multiple deep well plates complete the entire extraction process together, one at a time Extraction of 96 samples.
  • Such magnetic separation devices require the cooperation of a liquid workstation to complete the dispensing of samples and reagents when fully automating the entire extraction process, or by preloading reagents in a deep well plate and manually loading the samples.
  • the above magnetic pen separation method has many shortcomings: 1) It is not convenient to track the processing of each independent sample. Due to the batch processing method, a certain number of samples are processed at the same time in each stage, and the failure conditions of each sample individual in the batch can not be correspondingly responded and processed in the process; 2) The number of samples is not In the case of certain determination, the use efficiency is not high. According to the number of magnetic pens and the design of the corresponding reaction container, it is necessary to design the processing flux to obtain the best performance. If the number of samples is less than the design throughput, it will cause waste of consumables; if the number of samples is greater than the design throughput, it will need to be re-run again.
  • Another magnetic bead nucleic acid extraction device is a magnetic separation device mounted on a liquid processing workstation.
  • Such magnetic separation devices generally consist of a square plate and a rod-shaped magnet perpendicular to the plane of the square plate in an array of distances or a strip-shaped magnet parallel to the plane.
  • the magnetic separating device can adsorb the magnetic particles in each hole of the deep hole plate on the side wall, and the reagents of the liquid working station are used to inject or transfer the reagents to complete the processing of different reagents, thereby completing one in the same hole.
  • the nucleic acid extraction process of the sample is a magnetic separation device mounted on a liquid processing workstation.
  • the object of the present invention is to solve the automation problem of applying the in vitro diagnostic reagent for reagent distribution and nano magnetic particle separation in the target substance extraction and detection process, and to provide a device for automatically dispensing the reagent and separating the nano magnetic particles.
  • the reagent can be distributed and reacted for independent samples, and the magnetic nanoparticles adsorbing the target substance can be separated into solid-liquid two-phase, which can be conveniently applied to the automation of sample processing in the in vitro diagnostic process using magnetic nanoparticles. .
  • the present invention provides a magnetic bead nucleic acid extraction device comprising a disk turntable in the shape of an inverted disk, an arc arch bridge base directly below the disk turntable, connecting the disk turntable and the circular arc a pillar in the vertical direction of the arch bridge base and located in the center of the disc, a magnet structural unit and a sample disposed on the disc turntable Reaction unit
  • the disc turret includes a top plate, and the top plate is sequentially provided with a guide rod insertion hole and a sample reaction unit jack from a circumferential direction of the center of the circle;
  • the arc arch bridge base includes a bottom plate disposed at a lowermost portion and an arc strut plate disposed on the bottom plate, wherein the arc strut plate corresponds to a position of the guide bar insertion hole in a radial direction of the disk, and the circle
  • the arc strut includes a raised area for lifting the guide rod and the magnet and a recessed area for guiding the rod and the magnet to fall back;
  • the pillar is disposed above the bottom plate
  • the magnet structure unit includes a vertically disposed guide rod, a magnet, and a magnet holder for driving the magnet to move up and down synchronously when the guide rod moves up and down; the magnet holder and the magnet are disposed under the top plate, and a magnet in the radial direction of the disc is disposed between the guide rod and the pillar;
  • the sample reaction unit includes a sample tube holder plate that can be placed on the sample reaction unit receptacle, a sample tube disposed downward from the sample tube holder plate and adjacent to the magnet in the radial direction of the disk.
  • the guide rod insertion hole and the sample reaction unit insertion hole are each 24, and are evenly arranged in the circumferential direction of the top plate.
  • both the raised area and the recessed area are two, and are evenly spaced in the circumferential direction of the bottom plate.
  • the sample reaction unit includes a sample tube and a reserve sample tube disposed at both ends in the radial direction of the disk, and a reserve tube disposed in the middle.
  • the spare tube is used, for example, to contain waste liquid during nucleic acid extraction.
  • the sample tube is a large, small, and small cone tube.
  • the magnet is a strip magnet disposed vertically upward or obliquely upward.
  • the sample reaction unit further includes a shield frame disposed above the sample tube holder plate for preventing cross-infection of the sample between the sample reaction units.
  • the disc turret further comprises a rim extending downward along the top plate, and the rim is a circular arc-shaped rim or a rim surrounded by a plurality of rectangular plates.
  • the bottom plate is a circular bottom plate or a hollow annular bottom plate.
  • the length of the magnet is 1/2 to 1 times the length of the sample tube, for example, 2/3 times.
  • the top plate is sequentially provided with a guide rod insertion hole, a sample reaction unit insertion hole and a pillar insertion hole from a circumferential direction of the center of the circle.
  • FIG. 1 is a schematic view showing the overall structure of a magnetic bead nucleic acid extraction device of the present invention
  • FIG. 1a and 1b are left and right views of Fig. 1;
  • Figure 1c is a plan view of Figure 1;
  • Figure 1d is a front view of Figure 1;
  • FIG. 2 is a schematic view showing the overall structure of the magnet structural unit of FIG. 1;
  • Figure 2a is a front view of Figure 2;
  • FIG. 3 is a schematic view showing the overall structure of the sample reaction unit of FIG. 1;
  • Figure 3a is a front view of Figure 3;
  • Figure 3b is a top view of Figure 3.
  • 1 is a disc turntable
  • 11 is a top plate
  • 111 is a guide rod jack
  • 112 is a sample reaction unit jack
  • 113 is a pillar jack
  • 12 is a rim
  • 2 is an arc arch bridge base
  • 22 is a circular struts
  • 221 is a raised area
  • 222 is a grooved area
  • 3 is a pillar
  • 4 is a magnet structural unit
  • 41 is a guiding rod
  • 42 is a magnet bracket
  • 43 is a magnet
  • 5 is a sample reaction
  • 51 is a sample tube
  • 52 is a spare tube
  • 53 is a sample tube support plate
  • 54 is a reserved sample tube
  • 55 is a shield frame.
  • the present invention provides a magnetic bead nucleic acid extraction device, which comprises a disk turntable in the shape of an inverted disk, an arc arch bridge base directly below the disk turntable, and the disk is connected a turret and an arc arch bridge base, a struts disposed in a vertical direction at a center of the disc, a magnet structural unit disposed on the disc turret, and a sample reaction unit; wherein the disc turret includes a top plate, the top plate
  • the guide rod insertion hole, the sample reaction unit insertion hole and the pillar insertion hole 113 are sequentially opened from the circumference to the center of the circle;
  • the disc turntable further includes a circumference extending downward along the top plate, and the circumference 12 is a circle
  • the arc-plate type rim or the top surface formed by the connection of a plurality of rectangular plates is a regular polygonal rim.
  • the arc arch bridge base includes a bottom plate disposed at a lowermost portion and an arc strut plate disposed on the bottom plate, wherein the arc strut plate corresponds to a position of the guide bar insertion hole in a radial direction of the disk, and the circle
  • the arc strut includes a raised area for lifting the guide rod and the magnet and a recessed area for the guide rod and the magnet to fall back; the raised area and the recessed area are both, and in the circumferential direction of the bottom plate Evenly spaced.
  • the raised regions include a B region and a D region, and the recessed regions include an A region and a C region.
  • the bottom plate is a circular bottom plate.
  • the post is disposed above the bottom plate.
  • the magnet structure unit includes a vertically disposed guide rod, a magnet, and a magnet holder for driving the magnet to move up and down synchronously when the guide rod moves up and down; the magnet holder and the magnet are disposed under the top plate, and A magnet in the radial direction of the disc is disposed between the guide rod and the strut; the magnet is a strip magnet disposed vertically upward or obliquely upward.
  • the sample reaction unit includes a sample tube holder plate that can be placed on the sample reaction unit receptacle, a sample tube disposed downward from the sample tube holder plate and adjacent to the magnet in the radial direction of the disk.
  • the sample reaction unit includes a sample tube and a reserved sample tube disposed at both ends in the radial direction of the disk, and a spare tube disposed in the middle.
  • the spare tube 52 is used, for example, to contain waste liquid during nucleic acid extraction.
  • the sample tube is a large and small conical tube.
  • the sample reaction unit further includes a shield frame 55 disposed above the sample tube holder plate for preventing cross-infection of the sample between the sample reaction units.
  • the guide rod insertion hole and the sample reaction unit insertion hole are each 24, and are evenly arranged in the circumferential direction of the top plate.
  • the length of the magnet is 2/3 times the length of the sample tube.
  • the magnetic bead nucleic acid extraction device is used and moved in such a manner that when the disk turret 1 rotates, a certain magnet structural unit 4 and the sample reaction unit 5 on the disk turret 1 are rotated to the raised area.
  • the raised area 221 pushes up the guiding rod 41, and the guiding rod 41 drives the magnet 43 to move upward, and the magnet is close to the sample tube 51, thereby applying a magnetic field to the sample tube;
  • Magnet structure unit 4 When the sample reaction unit 5 is rotated to a position corresponding to the recessed portion 222, the guide rod 41 is lowered downward, and the guide rod 41 drives the magnet 43 to move downward, and the magnet is away from the sample in the vertical direction. Tube 51, thereby removing the magnetic field from the sample tube.
  • the position of the magnetic field applied to the outer wall of the sample tube 51 can also be changed by designing the height of the raised portion 221, thereby controlling the position of the nanomagnetic beads at the enriched height of the inner wall of the sample tube 51.
  • the disk turret 1 is rotated from the position corresponding to the A zone to the B zone, the C zone and the D zone, and the nucleic acid cleavage and washing process is realized after one cycle of completion.
  • the disc turret is turned back to the A zone, the nucleic acid-magnetic bead complex can be resuspended by adding the PCR reaction solution and transferred to a PCR tube, and then detected by a PCR instrument.
  • the device provided by the invention enables pipelined automated nucleic acid extraction.

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Abstract

一种磁珠法核酸提取装置，包括呈倒扣圆盘状的圆盘转台、位于圆盘转台正下方的圆弧拱桥底座、连接圆盘转台和圆弧拱桥底座且位于圆盘中央的沿竖直方向设置的支柱、设置在圆盘转台上的磁铁结构单元和样本反应单元。圆盘转台包括顶板，顶板上从其圆周向圆心方向依次开设有导向杆插孔和样本反应单元插孔。圆弧拱桥底座包括设置在最下方的底板和设置在底板上的圆弧撑板，圆弧撑板在圆盘的径向上与所述导向杆插孔位置对应，且圆弧撑板包括用于顶起导向杆和磁铁的凸起区和用于导向杆和磁铁回落的凹槽区。本发明提供的装置能实现流水线式的自动化核酸提取。

Description

一种磁珠法核酸提取装置
本申请要求于2015年1月14日提交中国专利局、申请号为201510017498.6、发明名称为“一种磁珠法核酸提取装置”的中国专利申请的优先权，其全部内容通过引用结合在本申请中。
技术领域
本发明涉及核酸提取装置领域，具体涉及一种磁珠法核酸提取装置
背景技术
在传染性和遗传性疾病的诊断过程中，分子水平的检测(如核酸检测)能显著地缩短检测窗口期和提高检测灵敏度。通常，从标本中获得的分析物的质量(如纯度等)是影响检测结果的重要因素。就核酸检测而言，从标本中提取的核酸的量和纯度，直接影响后续PCR(聚合酶链式反应)检测的结果。
核酸的提取一般经历下述几个步骤：1)包被核酸的膜质的裂解及核酸的释放；2)蛋白的去除；3)核酸的收集。经典的方法为煮沸裂解法和酚-氯仿提取法。煮沸裂解法先在样本中加入沉淀剂后离心弃上清，以去除小分子抑制物并沉淀病毒颗粒；然后加入裂解液后煮沸，以释放DNA及沉淀残留蛋白等大分子抑制物，离心取上清即为得到的DNA。酚-氯仿提取法先在样本中加入裂解液，再加入氯仿后离心，以释放RNA并使之与蛋白层分离，再将上清液加入到异丙醇中以萃取RNA，离心后弃上清，加入乙醇洗涤后即为得到的RNA。这两种方法由于步骤比较繁琐，涉及到多次离心或加热，不便于实现自动化且手工劳动强度大。在此基础上，有一种采用膜技术进行核酸提取的方法，简化了操作，可以实现自动化。然而，在利用膜对核酸进行洗涤和收集时仍然需要离心机来完成，这也是该方法的一个较大局限。
随着材料学的发展，一种纳米尺寸的微粒(磁珠)被用于核酸的提取过程。这种 顺磁性纳米颗粒可以特异或非特异地吸附核酸，并利用磁场可以将核酸从水相中方便地分离出来，达到提取核酸的目的。这种材料一经采用，便引起了自动化核酸提取技术的研发。随即出现了多种基于纳米磁性微粒的自动化核酸提取技术和装置。例如，核酸提取的步骤具体包括：在含核酸的样本中，1)裂解：加入裂解液，样本裂解出核酸后，2)结合：加入磁珠，磁珠特异地结合核酸，3)洗涤：加入洗涤液，通过洗涤步骤把核酸上的蛋白质等杂质去除，4)洗脱：加入洗脱液将磁珠和核酸分开，5)分离：在磁场条件下分离富集磁珠，得到纯化的核酸。然而，对应该核酸提取的步骤却有相当多的操作步骤。具体地，先在烧杯(或样本管)中放入裂解液、磁珠和样本，磁棒(棒状磁铁)和套在磁棒一头的一次性外套(Tip)配合使用置入含上述溶液的烧杯中，先使磁棒和其外套缓慢地上下运动以收集磁珠和核酸的结合体，然后将磁棒连同外套转入另一种溶液(洗涤液)中，先把磁棒取出，磁场消失，外套在洗涤液中快速地做上下运动，磁珠和核酸的结合物就进入溶液中。然后磁棒又套好外套去收集磁珠和核酸的结合物，再在洗脱液中将磁棒和外套分开。得到含磁珠和核酸的溶液再回到烧杯中的方式进行磁珠收集，得到纯净的核酸溶液。
也就是说，棒状磁铁(Magnet)和一次性外套(Tip)的配合使用，可以完成将磁珠从一种试剂到另一种试剂的转移。在具体的实例中，主要采取将上述棒状磁铁进行阵列(2列*8行、4列*8行或者12列*8行)，一次性外套的阵列方式则是1*8或12*8，反应管则采用96孔深孔板。从工作方式上，可以分成2种：其一，采用2*8或4*8的磁棒阵列和1*8的一次性外套，在一块深孔板中完成16个或32个样本的提取过程。其二，采用12*8的磁棒阵列和12*8的一次性外套，在一块深孔板中完成一个提取步骤(一种试剂)，多块深孔板一起完成整个提取过程，一次可以完成96个样本的提取。
这类磁性分离装置在实现整个提取过程的全面自动化时，需要液体工作站的配合来完成样本和试剂的分配；或者采取在深孔板内预装试剂，手工装载样本来实现。此 外，上述磁笔式分离方法还有很多不足之处：1)不便于跟踪每个独立样本的处理过程。由于采用批处理方式，一定个数的样本同时进行各阶段处理，批内的每个样本个体在各阶段发生的失效情况无法在处理过程中得到相应的响应和处理；2)在样本个数不确定的场合使用效率不高，根据磁笔个数和相应反应容器的设计，必须在设计处理通量下才能获得最佳的性能。若样本个数少于设计通量，则将造成耗材的浪费；若样本个数大于设计通量，则需要重新运行一遍。
另一种磁珠法核酸提取装置是搭载在液体处理工作站上磁性分离装置。这类磁性分离装置一般由一块方板和按照一定距离阵列的垂直于方板平面的棒状磁铁或是平行于平面的条状磁铁组成。这种磁性分离装置可以将深孔板每个孔内的磁性微粒吸附在侧壁上，借助液体工作站的移液机械臂将试剂注入或转移来完成不同试剂的处理，进而在同一孔内完成一个样本的核酸提取过程。
因此，鉴于上述现有技术中的磁珠法核酸提取装置均为手动或半自动装置，只能做到核酸提取的单独处理或分批处理。因此，本领域需提供一种全自动的磁珠法核酸提取装置，使用该装置能连续化地进行核酸提取。
发明内容
本发明的目的在于解决应用体外诊断试剂进行靶物质提取和检测过程中试剂分配和纳米磁性微粒分离的自动化问题，提供一种便于自动分配试剂并分离纳米磁性微粒的装置。应用该装置，可以为独立样本进行试剂的分配、反应，并将吸附有靶物质的磁性纳米微粒进行固液两相分离，可以方便地应用于使用磁性纳米颗粒的体外诊断过程中样本处理的自动化。
因此，本发明提供一种磁珠法核酸提取装置，所述装置包括呈倒扣圆盘状的圆盘转台、位于圆盘转台正下方的圆弧拱桥底座、连接所述圆盘转台和圆弧拱桥底座且位于圆盘中央的沿竖直方向设置的支柱、设置在所述圆盘转台上的磁铁结构单元和样本 反应单元；
其中，所述圆盘转台包括顶板，所述顶板上从其圆周向圆心方向依次开设有导向杆插孔和样本反应单元插孔；
所述圆弧拱桥底座包括设置在最下方的底板和设置在底板上的圆弧撑板，所述圆弧撑板在圆盘的径向上与所述导向杆插孔位置对应，且所述圆弧撑板包括用于顶起导向杆和磁铁的凸起区和用于导向杆和磁铁回落的凹槽区；
所述支柱设置在所述底板上方；
所述磁铁结构单元包括竖直设置的导向杆、磁铁和用于在导向杆上下运动时带动所述磁铁同步上下运动的磁铁支架；所述磁铁支架和磁铁均设置在所述顶板下方，且在圆盘径向上磁铁设置在所述导向杆与所述支柱之间；
所述样本反应单元包括能放置在所述样本反应单元插孔上的样本管支座板、从样本管支座板向下设置且在圆盘径向上邻近磁铁的样本管。
优选地，所述导向杆插孔、样本反应单元插孔各为24个，且均在所述顶板的圆周方向上均匀排列。
优选地，所述凸起区和凹槽区均为2个，且在所述底板圆周方向上均匀间隔排列。
在一种具体的实施方式中，所述样本反应单元包括沿圆盘径向设置在两端的样本管和预留样本管，以及设置在中间的备用管。所述备用管例如用于盛装核酸提取过程中的废液。本领域技术人员可理解的，当所述样本反应单元沿圆盘径向水平旋转180°再放置在样本反应单元插孔上时，原来的预留样本管变为样本管。
在一种具体的实施方式中，所述样本管为上大下小的圆锥体管。
优选地，所述磁铁为竖直向上设置或斜向上设置的条形磁铁。
在一种具体的实施方式中，所述样本反应单元还包括设置在样本管支座板上方的用于防止各样本反应单元间样本交互感染的屏蔽框。
优选地，所述圆盘转台还包括沿顶板向下方延伸的围边，且所述围边为圆弧板型围边或由多块矩形板首尾连接形成的俯视面为正多边形的围边。
在一种具体的实施方式中，所述底板为圆形底板或中空的圆环形底板。
优选地，所述磁铁的长度为所述样本管长度的1/2～1倍，例如为2/3倍。
优选地，所述顶板上从其圆周向圆心方向依次开设有导向杆插孔、样本反应单元插孔和支柱插孔。
附图说明
构成本申请的一部分的说明书附图用来提供对本发明的进一步理解，本发明的示意性实施例及其说明用于解释本发明，并不构成对本发明的不当限定。在附图中：
图1为本发明中磁珠法核酸提取装置的整体结构示意图；
图1a和图1b为图1的左向和右向视图；
图1c为图1的俯视图；图1d为图1的主视图；
图2为图1中的磁铁结构单元的整体结构示意图；
图2a为图2的主视图；
图3为图1中的样本反应单元的整体结构示意图；
图3a为图3的主视图；
图3b为图3的俯视图。
在上述附图中，1为圆盘转台，11为顶板，111为导向杆插孔，112为样本反应单元插孔，113为支柱插孔，12为围边，2为圆弧拱桥底座，21为底板，22为圆弧撑板，221为凸起区，222为凹槽区，3为支柱，4为磁铁结构单元，41为导向杆，42为磁铁支架，43为磁铁，5为样本反应单元，51为样本管，52为备用管，53为样本管支座板，54为预留样本管，55为屏蔽框。
具体实施方式
本发明的目的是通过以下技术方案实现的。以下实施例用于说明本发明，但并不用于限定本发明的保护范围。
如图1所示，本发明提供一种磁珠法核酸提取装置，所述装置包括呈倒扣圆盘状的圆盘转台、位于圆盘转台正下方的圆弧拱桥底座、连接所述圆盘转台和圆弧拱桥底座且位于圆盘中央的沿竖直方向设置的支柱、设置在所述圆盘转台上的磁铁结构单元和样本反应单元；其中，所述圆盘转台包括顶板，所述顶板上从其圆周向圆心方向依次开设有导向杆插孔、样本反应单元插孔和支柱插孔113；所述圆盘转台还包括沿顶板向下方延伸的围边，且所述围边12为圆弧板型围边或由多块矩形板首尾连接形成的俯视面为正多边形的围边。
所述圆弧拱桥底座包括设置在最下方的底板和设置在底板上的圆弧撑板，所述圆弧撑板在圆盘的径向上与所述导向杆插孔位置对应，且所述圆弧撑板包括用于顶起导向杆和磁铁的凸起区和用于导向杆和磁铁回落的凹槽区；所述凸起区和凹槽区均为2个，且在所述底板圆周方向上均匀间隔排列。在图1d中，所述凸起区包括B区和D区，而所述凹槽区包括A区和C区。所述底板为圆形底板。
所述支柱设置在所述底板上方。所述磁铁结构单元包括竖直设置的导向杆、磁铁和用于在导向杆上下运动时带动所述磁铁同步上下运动的磁铁支架；所述磁铁支架和磁铁均设置在所述顶板下方，且在圆盘径向上磁铁设置在所述导向杆与所述支柱之间；所述磁铁为竖直向上设置或斜向上设置的条形磁铁。
所述样本反应单元包括能放置在所述样本反应单元插孔上的样本管支座板、从样本管支座板向下设置且在圆盘径向上邻近磁铁的样本管。所述样本反应单元包括沿圆盘径向设置在两端的样本管和预留样本管，以及设置在中间的备用管。所述备用管52例如用于盛装核酸提取过程中的废液。所述样本管为上大下小的圆锥体管。所述样本反应单元还包括设置在样本管支座板上方的用于防止各样本反应单元间样本交互感染的屏蔽框55。
其中，所述导向杆插孔、样本反应单元插孔各为24个，且均在所述顶板的圆周方向上均匀排列。磁铁的长度为所述样本管长度的2/3倍。
上述磁珠法核酸提取装置的使用方法和运动方式为：所述圆盘转台1转动时，当圆盘转台1上的某个磁铁结构单元4和样本反应单元5转动到与所述凸起区221相应的位置时，凸起区221将所述导向杆41向上顶起，导向杆41带动所述磁铁43向上运动，磁铁靠近所述样本管51，从而对样本管中施加磁场；当所述磁铁结构单元4 和样本反应单元5转动到与所述凹槽区222相应的位置时，所述导向杆41向下回落，导向杆41带动所述磁铁43向下运动，磁铁在竖直方向上远离所述样本管51，从而样本管中撤去磁场。
在本发明中，还可以通过对凸起区221的高度的设计而改变施加在所述样本管51外侧壁的磁场的位置，从而控制纳米磁珠在样本管51内壁的富集高度位置。
图1d中，圆盘转台1从与A区相应的位置转动到B区、C区和D区，完成循环一周后则实现了核酸的裂解和洗涤过程。当圆盘转台转回A区时，此时可以加入PCR反应液重悬核酸-磁珠复合物并将其转移至PCR管中，再使用PCR扩增仪进行检测。
本发明提供的装置能实现流水线式的自动化核酸提取。
以上是本发明的核心思想，为了使本领域技术人员更好地理解本发明方案，下面将结合本发明实施例中的附图，对本发明实施例中的技术方案进行清除、完整地描述，显然，所述描述的实施例仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。

Claims (10)

  1. 一种磁珠法核酸提取装置，所述装置包括呈倒扣圆盘状的圆盘转台、位于所述圆盘转台正下方的圆弧拱桥底座、连接所述圆盘转台和圆弧拱桥底座且位于所述圆盘转台中央并沿竖直方向设置的支柱、设置在所述圆盘转台上的磁铁结构单元和样本反应单元；
    其中，所述圆盘转台包括顶板，所述顶板上从其圆周向圆心方向依次开设有导向杆插孔、样本反应单元插孔和支柱插孔；
    所述磁铁结构单元包括竖直设置的导向杆、磁铁和用于在导向杆上下运动时带动所述磁铁同步上下运动的磁铁支架；所述磁铁支架和磁铁均设置在所述顶板下方，且在圆盘转台径向上磁铁设置在所述导向杆与所述支柱之间；
    所述样本反应单元包括能放置在所述样本反应单元插孔上的样本管支座板、从样本管支座板向下设置且在圆盘转台径向上邻近所述磁铁的样本管；
    所述圆弧拱桥底座包括设置在最下方的底板和设置在所述底板上的圆弧撑板，所述圆弧撑板在圆盘转台的径向上与所述导向杆插孔位置对应，且所述圆弧撑板包括用于顶起所述导向杆的凸起区和用于导向杆回落的凹槽区。
  2. 根据权利要求1所述的核酸提取装置，其特征在于，所述导向杆插孔、样本反应单元插孔各为24个，且在所述顶板的圆周方向上呈环形阵列。
  3. 根据权利要求1所述的核酸提取装置，其特征在于，所述凸起区和凹槽区均为2个，且在所述底板圆周方向上间隔阵列。
  4. 根据权利要求1所述的核酸提取装置，其特征在于，所述样本反应单元包括沿圆盘转台径向依次设置的样本管、备用管和预留样本管。
  5. 根据权利要求1所述的核酸提取装置，其特征在于，所述样本管为上大下小的圆锥体管。
  6. 根据权利要求1所述的核酸提取装置，其特征在于，所述磁铁为竖直向 上设置或斜向上设置的条形磁铁。
  7. 根据权利要求1～6中任一项所述的核酸提取装置，其特征在于，所述样本反应单元还包括设置在样本管支座板上方的用于防止各样本反应单元间样本交互感染的屏蔽框。
  8. 根据权利要求1～6中任一项所述的核酸提取装置，其特征在于，所述圆盘转台还包括沿顶板向下方延伸的围边，且所述围边为圆弧板型围边或由多块矩形板首尾连接形成的俯视面为正多边形的围边。
  9. 根据权利要求1～6中任一项所述的核酸提取装置，其特征在于，所述底板为圆形底板或中空的圆环形底板。
  10. 根据权利要求1～6中任一项所述的核酸提取装置，其特征在于，所述磁铁的长度为所述样本管长度的1/2～1倍。
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