WO2020037684A1 - 细胞及生物分子分离提取装置及测试系统 - Google Patents

细胞及生物分子分离提取装置及测试系统 Download PDF

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Publication number
WO2020037684A1
WO2020037684A1 PCT/CN2018/102346 CN2018102346W WO2020037684A1 WO 2020037684 A1 WO2020037684 A1 WO 2020037684A1 CN 2018102346 W CN2018102346 W CN 2018102346W WO 2020037684 A1 WO2020037684 A1 WO 2020037684A1
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Prior art keywords
magnetic beads
group
cells
biological sample
magnetic
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Ceased
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PCT/CN2018/102346
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English (en)
French (fr)
Inventor
赵静
江媛
章文蔚
赵霞
陈奥
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BGI Shenzhen Co Ltd
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BGI Shenzhen Co Ltd
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Priority to PCT/CN2018/102346 priority Critical patent/WO2020037684A1/zh
Priority to US17/267,452 priority patent/US12305166B2/en
Priority to CN201880095817.8A priority patent/CN112823202B/zh
Publication of WO2020037684A1 publication Critical patent/WO2020037684A1/zh
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • 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
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Definitions

  • the invention relates to the field of biotechnology, in particular to a cell and biomolecule separation and extraction device and a sequencing system.
  • Blood is composed of plasma, white blood cells, platelets, and red blood cells, and blood samples are the easiest to obtain the least invasive biopsy samples in clinical diagnosis. If it is a tumor patient, its white blood cells may be mixed with CTCs (circulating tumor cells) and CTM (circulating tumor microemboli), and its plasma may contain free ctDNA (circulating tumor DNA). ). The gene information of ctDNA, CTC and CTM is very important for the auxiliary diagnosis of tumors, the monitoring and monitoring of recurrence and metastasis, the evaluation of curative effect and the detection of drug resistance. Secondly, there are T cells and B cells in white blood cells, which are of great significance for studying autoimmune diseases and inflammatory responses.
  • gDNA genomic DNA, gene DNA
  • mRNA messenger RNA
  • miRNA miRNA
  • the invention provides a device for separating and extracting cells and biomolecules, including a first separating device.
  • the first separating device includes a sleeve for containing a biological sample solution, and the sleeve is provided with a channel for the biological sample solution to flow out. Holes, the through holes have different pore diameters; and multiple sets of magnetic beads, each set of magnetic beads used to identify and combine different cells or molecules in the biological sample, where the size of each group of magnetic beads is different The plurality of sets of magnetic beads respectively flow out from the through holes with corresponding pore diameters.
  • the sleeve includes an inner tube and an outer tube sleeved on the inner tube.
  • a first bead hole is provided on a peripheral wall of the inner tube, and a first tube hole is provided on a peripheral wall of the outer tube.
  • the first separating device further includes a first driving member, and the outer tube and the inner tube are capable of rotating relative to each other.
  • the first driving member is connected to the outer tube and the inner tube.
  • One is connected to drive one of the outer tube and the inner tube to rotate relative to the other, so as to change the overlapping area of the second bead hole and the corresponding first bead hole, thereby forming different pore sizes. Mentioned through holes.
  • the first separating device further includes a controller, the controller is connected to the first driving member, and is configured to control the first driving member to drive the outer tube or the inner tube to rotate.
  • the sleeve further includes a guide tube, and a guide hole is formed at the bottom of the outer tube, and the guide tube is in communication with the guide hole.
  • the first separation device further includes a magnetic field generator for stirring the biological sample solution contained in the sleeve, and the magnetic field generator emits a magnetic field to act on the inside of the sleeve;
  • the first separating device further includes a second driving member connected to the magnetic field generator and configured to drive the magnetic field generator into and out of the sleeve.
  • the multiple sets of magnetic beads include:
  • a third group of magnetic beads for identifying and binding T cells in the biological sample
  • a fourth group of magnetic beads for identifying and binding gDNA in the biological sample
  • a fifth set of magnetic beads for identifying and binding mRNA in said biological sample A fifth set of magnetic beads for identifying and binding mRNA in said biological sample.
  • a sixth group of magnetic beads for identifying and binding miRNAs in the biological sample
  • the second driver drives the magnetic field The generator enters the sleeve; after the magnetic field generator adsorbs the first group of magnetic beads, the second group of magnetic beads, and the third group of magnetic beads, the second driving member drives the magnetic field generator to move out of the station.
  • the magnetic field generated by the magnetic field generator is an electromagnetic field.
  • the fourth group of magnetic beads binds gDNA, the fifth group of magnetic beads binds mRNA, and the sixth group of magnetic beads binds miRNA.
  • the driving member drives the magnetic field generator to enter the sleeve again, and the electromagnetic field generated by the magnetic field generator is removed, so that the first group of magnetic beads, the second group of magnetic beads, and the third group are adsorbed on the magnetic field generator.
  • the set of magnetic beads is released in the sleeve.
  • it further includes a recovery device for recovering the first group of magnetic beads, the second group of magnetic beads, the third group of magnetic beads, the fourth group of magnetic beads, and the fifth group of magnetic beads flowing out of the sleeve. And the sixth set of magnetic beads;
  • the recovery device is provided with a plurality of channels, and the plurality of channels communicate with the plurality of through holes one by one, respectively.
  • it further includes a second separation device for separating CTCs and CTM in the biological sample;
  • the second separation device is a filter membrane.
  • a cell and biomolecule testing system includes any one of the above-mentioned cell and biomolecule separation and extraction devices, a microfluidic system, and a gene sequencing system.
  • the microfluidic system is combined with the cell and biomolecule separation and extraction device.
  • the gene sequencing system is combined with the microfluidic system.
  • the separation and extraction device provided by the present invention combines cells and biomolecules of different sizes through magnetic beads of different sizes, so that magnetic beads of different sizes can flow out through the through holes for separation, so as to separate a variety of cells and organisms in the biological sample.
  • the molecules are separated at the same time.
  • the cell and biomolecule testing system provided by the present invention realizes automation and improves processing efficiency.
  • FIG. 1 is a functional module diagram of a separation and extraction device provided by the present invention.
  • FIG. 2 is a schematic structural diagram of the sleeve shown in FIG. 1.
  • FIG. 3 is a functional module diagram of a test system provided by the present invention.
  • Separation and extraction device 100 First separation device 10 casing 12 Entrance 121 Through hole 124
  • a component when a component is considered to be “connected” to another component, it may be directly connected to another component or a centered component may exist at the same time.
  • a component When a component is considered to be “set on” another component, it can be directly set on another component or a centered component may exist at the same time.
  • FIG. 1 is a functional block diagram of a separation and extraction apparatus 100 according to a first embodiment of the present invention.
  • the separation and extraction device 100 is used to separate and extract various cells, such as B cells and T cells, and various nucleic acid molecules, such as ct / cfDNA, gDNA, mDNA, and miRNA, from a biological sample.
  • the biological sample in the present invention is a blood sample. It may be understood that the biological sample may also be a sample obtained by processing other animals, plants, tissues, cells, and the like.
  • the separation and extraction device 100 includes a first separation device 10 for separating ct / cfDNA, B cells, T cells, ctDNA, gDNA, mDNA, and miRNA in a biological sample.
  • the first separation device 10 performs separation based on an immunomagnetic separation technology.
  • the first separation device 10 includes a sleeve 12 and a plurality of sets of magnetic beads.
  • FIG. 2 is a schematic structural diagram of the sleeve 12 shown in FIG. 1.
  • the sleeve 12 is used for containing a biological sample solution.
  • One end of the sleeve 12 has an inlet 121 through which the biological sample solution flows into the sleeve 12.
  • the sleeve 12 is provided with a plurality of through holes 124 for the biological sample solution flowing into the sleeve 12 to flow out. Specifically, the through hole 124 is opened on a peripheral wall of the sleeve 12.
  • Each group of magnetic beads is used to identify and bind different cells or molecules in the biological sample, wherein the size of each group of magnetic beads is different, and is used to bind cells or molecules with different sizes, and respectively from different pore sizes.
  • the through hole 124 flows out.
  • the plurality of groups of magnetic beads includes a first group of magnetic beads, a second group of magnetic beads, a third group of magnetic beads, a fourth group of magnetic beads, a fifth group of magnetic beads, and a sixth group of magnetic beads.
  • the first set of magnetic beads is used for identifying and binding ct / cfDNA in the biological sample.
  • the second set of magnetic beads is used for identifying and binding B cells in the biological sample.
  • the third group of magnetic beads is used for identifying and binding T cells in the biological sample.
  • the fourth group of magnetic beads is used for identifying and binding gDNA in the biological sample.
  • the fifth group of magnetic beads is used for identifying and binding mRNA in the biological sample.
  • the sixth group of magnetic beads is used for identifying and binding miRNA in the biological sample.
  • each set of magnetic beads can include multiple magnetic beads of the same size.
  • the magnetic beads include surface-modified magnetic nanoparticles and specific molecules bound on the surface of the magnetic nanoparticles.
  • the surface chemical modification technology includes treatment methods such as hydroxylation, amination, and carboxylation, which are used to bond active groups on the surface of the magnetic nanoparticles to make them have good biocompatibility.
  • Ligand coupling enables recognition and binding of corresponding antigens, antibodies or nucleic acids.
  • the material of the magnetic nanoparticles is a ferromagnetic material or a paramagnetic material. Paramagnetic materials include materials such as aluminum, magnesium, and platinum that are only weakly attracted by strong magnets.
  • Ferromagnetic materials include nickel, cobalt, pure iron, iron alloys and other materials that are strongly attracted by magnets.
  • the specific molecule is a molecule capable of binding to ct / cfDNA, B cells, T cells, gDNA, mDNA, or miRNA.
  • specific molecules on magnetic beads used to bind B cells and T cells are immune antibodies, and specific molecules on magnetic beads used to bind ct / cfDNA, gDNA, mDNA, and miRNA are oligos. .
  • the sleeve 12 includes an inner tube 122 and an outer tube 123 sleeved on the inner tube 122.
  • the inner tube 122 and the outer tube 123 are substantially columnar. It can be understood that, in other embodiments, the inner tube 122 and the outer tube 123 may also be spherical, or have any other shape. Openings are formed at the same ends of the inner tube 122 and the outer tube 123, and the openings of the inner tube 122 and the outer tube 123 are substantially flush with each other.
  • the inner tube 122 is inserted into the outer tube 123 through an opening formed in the outer tube 123 and is received in the outer tube 123.
  • the opening of the inner tube 122 serves as an inlet 121 of the sleeve 12.
  • a plurality of first bead holes 125 are defined in a peripheral wall of the inner tube 122.
  • a plurality of second bead holes 126 are defined in a peripheral wall of the outer tube 123.
  • the positions of the plurality of second bead holes 126 are in one-to-one correspondence with the positions of the plurality of first bead holes 125.
  • a portion where each second bead hole 126 and a corresponding first bead hole 125 communicate with each other forms the communication hole.
  • 124 Hole 124.
  • portions of each second bead hole 126 and the corresponding first bead hole 125 overlapping each other collectively form the through hole 124.
  • the size of the second bead hole 126 and the size of the first bead hole 125 are substantially the same.
  • the first separating device 10 further includes a first driving member 14.
  • the first driving member 14 is connected to one of the inner pipe 122 and the outer pipe 123, and is used for driving one of the inner pipe 122 and the outer pipe 123 to rotate relative to the other to change
  • the overlapping area of the second bead hole 126 and the corresponding first bead hole 125 further changes the hole diameter of the through hole 124 so that the hole diameter of the through hole 124 is different from the first group of magnetic beads and the second hole.
  • the sizes of the group of magnetic beads, the third group of magnetic beads, the fourth group of magnetic beads, the fifth group of magnetic beads, and the sixth group of magnetic beads correspond.
  • the corresponding diameter of the through hole 124 and the size of the magnetic bead means that the diameter of the through hole 124 is approximately the same as the size of the corresponding cell or molecule-bound magnetic bead, or the size of the through hole 124
  • the pore size is larger than the size of the corresponding cell or molecule-bound magnetic beads but smaller than the size of other cells or molecules-bound magnetic beads.
  • the outer tube 123 is rotatably sleeved on the inner tube 122; the first driving member 14 is connected to the outer tube 123 for driving the outer tube 123 to rotate relative to the inner tube 122 .
  • the inner tube 122 is rotatably disposed in the outer tube 123; the first driving member 14 is connected to the inner tube 122 for driving the inner tube The tube 122 rotates relative to the outer tube 123.
  • the first driving member 14 drives the outer tube 123 to rotate, and changes the overlapping area of the second bead hole 126 and the corresponding first bead hole 125, so that the The pore diameter matches the size of the first set of magnetic beads bound with ct / cfDNA, so that only the first set of magnetic beads bound with ct / cfDNA flows out of the through hole 124, thereby separating one Magnetic beads.
  • the first driving member 14 drives the outer tube 123 to rotate, and changes the aperture of the through hole 124 so that only the second set of magnetic beads bound to the B cells can be transferred by the The through-hole 124 flows out; when T cells need to be separated, the first driving member 14 drives the outer tube 123 to rotate, and changes the aperture of the through-hole 124 so that only a third group of magnetic beads bound with T cells can be separated by The through-hole 124 flows out; when gDNA needs to be separated, the first driving member 14 drives the outer tube 123 to rotate, and changes the aperture of the through-hole 124 so that only the fourth set of magnetic beads bound with gDNA can be changed by The through hole 124 flows out; when mRNA is required to be separated, the first driving member 14 drives the outer tube 123 to rotate, and changes the aperture of the through hole 124 so that only the fifth group of magnetic beads bound with mRNA can be separated by The through-hole 124 flows out; when miRNA needs to be separated, the first driving
  • a plurality of through holes 124 with different apertures and controllable opening and closing can be directly opened on the sleeve 12.
  • a cover body capable of covering the through holes 124 is provided. The opening / closing of the body controls the opening and closing of the through hole 124.
  • the pore diameters of the plurality of through-holes 124 are respectively matched with the sizes of magnetic beads combined with ct / cfDNA, B cells, T cells, gDNA, mDNA, or miRNA.
  • the respective binding Magnetic beads with ct / cfDNA, B cells, T cells, gDNA, mDNA, and miRNA flow from the through hole 124 in order from small to large.
  • the number of the through holes 124 may be one or two, and the corresponding numbers of the first bead holes 125 and the second bead holes 126 are one or two, respectively.
  • the first separation device further includes a controller 16.
  • the controller 16 is connected to the first driving member 14 and is used to control the first driving member 14 to drive the outer tube 123 or the inner tube 122 to rotate, and then control to change the diameter of the through hole 124.
  • the controller 16 further controls the frequency at which the aperture of the through hole 124 changes.
  • the controller 16 controls the aperture of the through hole 124 from small to large, so that magnetic beads with different sizes can flow out of the through hole 124 separately in order from small to large.
  • the cannula 12 further includes a catheter 128.
  • a guide hole is provided at the bottom of the outer tube 123, and the conduit 128 is in communication with the guide hole, and is used for spilling a biological sample in a gap between the outer tube 123 and the inner tube 122. The solution was exported.
  • the first separation device 10 further includes a magnetic field generator 18 for generating a magnetic field.
  • the magnetic field generated by the magnetic field generator 18 acts on the inside of the inner tube 122, so that the magnetic beads are adsorbed on the magnetic field generator 18 under the force of the magnetic field.
  • the magnetic field generator 18 is used to stir the biological sample solution, so that the plurality of magnetic beads can smoothly flow out from the through hole 124.
  • the magnetic field generated by the magnetic field generator 18 is an electromagnetic field, for example, the magnetic field generator 18 is an electromagnetic rod. It can be understood that, in other embodiments, the magnetic field generated by the magnetic field generator 18 may also be a constant magnetic field, for example, the magnetic field generator 18 is a rhenium ferromagnetic rod.
  • the first separating device 10 further includes a second driving member 19.
  • the second driving member 19 is connected to the magnetic field generator 18 and is used to drive the magnetic field generator 18 into or out of the inner tube 122 and to drive the magnetic field generator 18 to stir the biological sample solution.
  • the second driving member 19 is further connected to the controller 16, and the controller 16 controls the second driving member 19 to drive the magnetic field generator 18.
  • the first set of magnetic beads, the second set of magnetic beads, and the third set of magnetic beads are injected into the biological sample solution, and the first set of magnetic beads binds ct / cfDNA, and the second set of magnetic beads
  • the second driver 19 drives the magnetic field generator 18 into the inner tube 122, and the first group of magnetic beads, the second group of magnetic beads, The group of magnetic beads and the third group of magnetic beads are attracted to the magnetic field generator 18 under the effect of the magnetic field force; the second driving member 19 drives the magnetic field generator 18 to move out of the inner tube 122.
  • the fourth group of magnetic beads, the fifth group of magnetic beads, and the sixth group of magnetic beads are injected into the biological sample solution, gDNA is bound to the fourth group of magnetic beads, and mRNA is bound to the fifth group of magnetic beads.
  • the sixth group of magnetic beads binds miRNA
  • the first group of magnetic beads bound with ct / cfDNA, the second group of magnetic beads bound with B cells, and the third group of magnetic beads bound with T cells are injected into the biological sample solution. in.
  • the magnetic field generated by the magnetic field generator 18 is an electromagnetic field
  • gDNA is bound to the fourth group of magnetic beads
  • mRNA is bound to the fifth group of magnetic beads
  • miRNA is bound to the sixth group of magnetic beads.
  • the driving member 19 drives the magnetic field generator 18 to enter the inner tube 122 again, and the electromagnetic field generated by the magnetic field generator 18 is removed, for example, by cutting off the power, so that the magnetic field generator 18 is combined with A first set of magnetic beads of ct / cfDNA, a second set of magnetic beads bound to B cells, and a third set of magnetic beads bound to T cells are released in the biological sample solution.
  • the magnetic field generator 18 When the magnetic field generated by the magnetic field generator 18 is a constant magnetic field, after the second driving member 19 drives the magnetic field generator 18 to move out of the inner tube 122, the magnetic field generator 18 is combined with The first set of magnetic beads of ct / cfDNA, the second set of magnetic beads bound to B cells, and the third set of magnetic beads bound to T cells are detached from the magnetic field generator 18 by an external force; The bead binds gDNA, the fifth group of magnetic beads binds mRNA, and after the sixth group of magnetic beads binds miRNA, the first group of magnetic beads bound to ct / cfDNA, the second group of magnetic beads bound to B cells, and A third set of magnetic beads bound with T cells is injected into the biological sample solution.
  • the separation and extraction device further includes a recovery device 30 for recovering the first group of magnetic beads, the second group of magnetic beads, the third group of magnetic beads, the fourth group of magnetic beads, the first group of magnetic beads flowing out of the sleeve 12.
  • a recovery device 30 for recovering the first group of magnetic beads, the second group of magnetic beads, the third group of magnetic beads, the fourth group of magnetic beads, the first group of magnetic beads flowing out of the sleeve 12.
  • the recovery device 30 is provided with a plurality of channels, and the plurality of channels are respectively in communication with the plurality of through holes 124 to recover the magnetic beads flowing out of the through holes 124 one by one. It can be understood that, in other embodiments, the recovery device 30 may be provided with only one channel, and the magnetic beads flowing out through the plurality of through holes 124 are recovered in the recovery device 30 through the channel.
  • the separation and extraction device 100 further includes a second separation device 40 for separating CTCs and CTM in the biological sample.
  • the second separation device 40 performs filtering based on a tumor cell size separation method (ISET, isolation by size).
  • ISET tumor cell size separation method
  • the second separation device 40 is a filter membrane, and the filter membrane is accommodated in the inner tube 122 of the sleeve 12 and divides the inner cavity of the inner tube 122 into upper and lower parts, Used to filter CTCs and CTM in the biological sample solution.
  • ct / cfDNA, B cells, T cells, ctDNA, gDNA, mDNA and miRNA were separated.
  • the filter membrane may also be disposed above or at the inlet of the sleeve 12.
  • the filter membrane may be any membrane that can be used for filtering, such as a polycarbonate (Track-etched) membrane, a micro-sieve membrane, a micro-grid (TEM grid) membrane, and the like.
  • the pore diameter of the filter membrane is 2.6-10um.
  • CTCs are generally larger than 25um, with a high nucleus and cytoplasm ratio, and abnormal morphology of chromosomes and nuclei.
  • CTM is a group of tumor cells that stick together, and its size is relatively large. After filtering through the filter membrane, CTCs and CTM with larger sizes are concentrated on the filter membrane, thereby separating CTCs and CTM. The separated CTCs and CTM can be removed for identification.
  • the second separating device 40 is a filtering device.
  • the second separation device 40 is in communication with the first separation device 10, and after the CTCs and CTMs in the biological sample are separated through the second separation device 40, the biological sample solution is injected into the first separation Device 10 for separating ct / cfDNA, B cells, T cells, ctDNA, gDNA, mDNA and miRNA in a biological sample.
  • the second separation device 40 includes a filter bottle and a filter membrane. The filter membrane is contained in the filter bottle, and the inner cavity of the filter bottle is divided into upper and lower parts for filtering CTCs and CTM in the biological sample solution.
  • the red blood cells in the blood sample are lysed, injected into the filter bottle, and filtered through the filter membrane and then flowed into the bottom of the filter bottle.
  • the filter bottle can be integrally formed, or can be assembled by detachably two parts.
  • the second separation device 40 further includes a pressure detection device and a pressure pump.
  • the pressure pump is in communication with a portion of the filter bottle below the filter membrane, and is used to adjust the pressure of the cavity in the filter bottle below the filter membrane, thereby regulating the biological sample passing through the filter membrane. Filtering speed.
  • the pressure detection device is used to detect the pressure of a cavity in the filter bottle below the filter membrane. The pressure pump automatically adjusts the pressure of the cavity in the filter bottle below the filter membrane according to the detection result of the pressure detection device.
  • the second separation device 40 can also be adsorbed by using a surface-functionalized membrane or a pipe system, thereby performing CTCs separation.
  • CTCs highly express EpCAM (epithelial cell adhesion molecule), and also express Cytokeratin's 4-6, 8, 10, 13 or 19 (cytokeratin 4-6, 8, 10, 13 or 19). Coupling anti-EpCAM, Cytokeratin's 4-6, 8, 10, 13 or 19 antibodies on the surface of the membrane or pipe. When biological samples flow through the surface of the membrane or pipe, the surface antigens of CTCs can be combined with the corresponding antibodies to achieve CTCs. Capture. Because CTCs do not express CD45, CTCs can also be isolated by CD45 negative screening. It can be understood that, in other embodiments, the second separation device 40 may also perform CTCs separation by using the method of Dean's vortex mechanics sorting method.
  • the separation and extraction device 100 provided by the present invention combines cells and biomolecules of different sizes through magnetic beads of different sizes, and drives the outer tube 123 to rotate through the first driving member 14 to adjust the aperture of the through hole 124, so that The beads can pass through the through holes 124 individually and sequentially, so as to separate multiple cells and biomolecules in the biological sample simultaneously.
  • FIG. 3 is a block diagram of a cell and biomolecule testing system 200 provided by the present invention.
  • the test system 200 includes any one of the above-mentioned separation and extraction device 100, a microfluidic system 210, and a gene sequencing system 230.
  • the microfluidic system 210 is combined with the separation and extraction device 100 and is used for constructing a sequencing library by using the nucleic acid molecules separated by the separation and extraction device 100.
  • the gene sequencing system 230 is combined with the microfluidic system 210 and is used to perform base sequence detection on different nucleic acid molecules isolated.

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Abstract

本发明提供一种细胞及生物分子分离提取装置，包括：套管，所述套管开设有供生物样品溶液流出的通孔，所述通孔具有不同孔径；以及多组磁珠，每组磁珠用于识别并结合所述生物样品溶液中的不同细胞或分子，其中各组磁珠之间的尺寸各不相同，所述多组磁珠分别从对应孔径的所述通孔流出。本发明还提供一种包括上述分离提取装置、微流控系统以及基因测序系统的细胞及生物分子测试系统。

Description

细胞及生物分子分离提取装置及测试系统 技术领域
本发明涉及生物技术领域，尤其涉及一种细胞及生物分子分离提取装置及测序系统。
背景技术
血液由血浆、白细胞、血小板和红细胞组成，血液样品是临床诊断中最易于获取造成创伤最小的活检样品。如果是肿瘤患者，其白细胞中可能混有CTCs(circulating tumor cells，循环肿瘤细胞)和CTM(circulating tumor microemboli，循环肿瘤微栓)，其血浆当中可能含有游离的ctDNA(circulating tumor DNA，循环肿瘤DNA)。ctDNA、CTC和CTM的基因信息对于肿瘤辅助诊断，术后复发转移与监测及疗效评估和耐药性检测具有非常重要的作用。其次，白细胞中还有T细胞和B细胞，其对于研究自身免疫类疾病及炎症反应具有很重要的意义。最后从剩余的白细胞和血小板中，可以通过裂解细胞获得gDNA(genomic DNA，基因DNA)，mRNA(messenger RNA，信使RNA)和miRNA(microRNA)，从而对血液提供者有一个全面的基因信息了解。然而，现有的血液样品提取分离试剂盒，一般针对一种或几种样品进行分离，较难对多种样品同时进行分离。
发明内容
鉴于上述状况，有必要提供一种能够同时提取分离多种样品的细胞及生物分子分离提取装置及测序系统。
本发明提供一种细胞及生物分子分离提取装置，包括第一分离装置，所述第一分离装置包括：套管，用于盛装生物样品溶液，所述套管开设有供生物样品溶液流出的通孔，所述通孔具有不同孔径；以及多组磁珠，每组磁珠用于识别并结合所述生物样品中的不同细胞或分子，其中各组磁珠之间的尺寸各不相同，所述多组磁珠分别从对应孔径的通孔流出。
作为一种优选方案，所述套管包括内管以及套设于所述内管上的外管，所述内管的周壁上开设有第一珠孔，所述外管的周壁上开设有第二珠孔，所述第二珠孔和所述第一珠孔的位置一一对应，每一第二珠孔和相应的第一珠孔相连通形成所述通孔。
作为一种优选方案，所述第一分离装置进一步包括第一驱动件，所述外管和所述内管能够相对转动，所述第一驱动件与所述外管和所述内管中的一个相连接，用于驱动所述外管和所述内管中的一个相对另一个转动，以改变所述第二珠孔与相应的第一珠孔的交叠面积，从而形成不同孔径的所述通孔。
作为一种优选方案，所述第一分离装置进一步包括控制器，所述控制器与所述第一驱动件相连接，用于控制所述第一驱动件驱动所述外管或内管转动。
作为一种优选方案，所述套管进一步包括导管，所述外管的底部开设有导引孔，所述导管与所述导引孔相连通。
作为一种优选方案，所述第一分离装置进一步包括磁场发生器，用于搅拌盛装于所述套管中的生物样品溶液，所述磁场发生器发出磁场作用于所述套管内部；
优选地，所述第一分离装置进一步包括第二驱动件，所述第二驱动件与所述磁场发生器相连接，用于驱动所述磁场发生器进入及移出 所述套管。
作为一种优选方案，所述多组磁珠包括：
第一组磁珠，用于识别并结合所述生物样品中的ct/cfDNA；
第二组磁珠，用于识别并结合所述生物样品中的B细胞；
第三组磁珠，用于识别并结合所述生物样品中的T细胞；
第四组磁珠，用于识别并结合所述生物样品中的gDNA；
第五组磁珠，用于识别并结合所述生物样品中的mRNA；以及
第六组磁珠，用于识别并结合所述生物样品中的miRNA；
优选地，在所述第一组磁珠结合ct/cfDNA，所述第二组磁珠结合B细胞，以及所述第三组磁珠结合T细胞后，所述第二驱动件驱动所述磁场发生器进入所述套管；在所述磁场发生器吸附所述第一组磁珠、第二组磁珠以及第三组磁珠后，所述第二驱动件驱动所述磁场发生器移出所述套管；在所述第四组磁珠结合gDNA，所述第五组磁珠结合mRNA，所述第六组磁珠结合miRNA后，结合有ct/cfDNA的第一组磁珠、结合有B细胞的第二组磁珠以及结合有T细胞的第三组磁珠被注入所述生物样品溶液中；
优选地，所述磁场发生器产生的磁场为电磁场，在所述第四组磁珠结合gDNA，所述第五组磁珠结合mRNA，所述第六组磁珠结合miRNA后，所述第二驱动件驱动所述磁场发生器再次进入所述套管，所述磁场发生器产生的电磁场被去除，使得吸附在所述磁场发生器上的第一组磁珠、第二组磁珠以及第三组磁珠被释放于所述套管中。
作为一种优选方案，进一步包括回收装置，用于回收由所述套管流出的第一组磁珠、第二组磁珠、第三组磁珠、第四组磁珠、第五组磁珠以及第六组磁珠；
优选地，所述回收装置设置有多个通道，所述多个通道分别与所 述多个通孔一一连通。
作为一种优选方案，进一步包括第二分离装置，用于分离所述生物样品中的CTCs和CTM；
优选地，所述第二分离装置为滤膜。
一种细胞及生物分子测试系统，包括上述任一细胞及生物分子分离提取装置、微流控系统以及基因测序系统，所述微流控系统与所述细胞及生物分子分离提取装置相结合，所述基因测序系统与所述微流控系统相结合。
本发明提供的分离提取装置通过不同尺寸的磁珠结合不同尺寸的细胞及生物分子，使得不同尺寸的磁珠通过所述通孔流出进行分离，从而对所述生物样品中的多种细胞及生物分子同时进行分离。本发明提供的细胞及生物分子测试系统，实现了自动化，提高了加工效率。
附图说明
图1为本发明提供的分离提取装置的功能模块图。
图2为图1所示套管的结构示意图。
图3为本发明提供的测试系统的功能模块图。
主要元件符号说明
分离提取装置 100
第一分离装置 10
套管 12
入口 121
通孔 124
内管 122
外管 123
第一珠孔 125
第二珠孔 126
第一驱动件 14
控制器 16
导管 128
磁场发生器 18
第二驱动件 19
回收装置 30
第二分离装置 40
测试系统 200
微流控系统 210
基因测序系统 230
如下具体实施方式将结合上述附图进一步说明本发明。
具体实施方式
在下面的描述中阐述了很多具体细节以便于充分理解本发明，但是本发明还可以采用其他不同于在此描述的其它方式来实施，本领域技术人员可以在不违背本发明内涵的情况下做类似应用，因此本发明不受下面公开的具体实施例的限制。
需要说明的是，当一个组件被认为是“连接”另一个组件，它可以是直接连接到另一个组件或者可能同时存在居中组件。当一个组件被认为是“设置于”另一个组件，它可以是直接设置在另一个组件上或者可能同时存在居中组件。
请参阅图1，图1为本发明第一实施方式提供的分离提取装置100的功能模块图。所述分离提取装置100用于分离提取生物样品中的多种细胞，例如B细胞和T细胞，以及多种核酸分子，例如ct/cfDNA、gDNA、mDNA和miRNA。具体地，本发明中的生物样品为血液样品，可以理解的是，所述生物样品还可以为其他动物、植物、组织、细胞等处理得到的样品。
所述分离提取装置100包括第一分离装置10，用于分离生物样品中的ct/cfDNA、B细胞、T细胞、ctDNA、gDNA、mDNA和miRNA。本实施例中，所述第一分离装置10基于免疫磁分离技术进行分离。具体地，所述第一分离装置10包括套管12以及多组磁珠。
请一并参阅图2，图2为图1所示套管12的结构示意图。所述套管12用于盛装生物样品溶液。所述套管12的一端具有入口121，生物样品溶液经由所述入口121流入所述套管12内。所述套管12上开设有多个通孔124，用于供流入所述套管12内的生物样品溶液流出。具体地，所述通孔124开设于所述套管12的周壁上。
每组磁珠用于识别并结合所述生物样品中不同的细胞或分子，其中各组磁珠之间的尺寸各不相同，用于结合具有不同尺寸的细胞或分子，并分别从不同孔径的通孔124流出。具体地，所述多组磁珠包括第一组磁珠、第二组磁珠、第三组磁珠、第四组磁珠、第五组磁珠以及第六组磁珠。所述第一组磁珠，用于识别并结合所述生物样品中的ct/cfDNA。所述第二组磁珠，用于识别并结合所述生物样品中的B细胞。所述第三组磁珠，用于识别并结合所述生物样品中的T细胞。所述第四组磁珠，用于识别并结合所述生物样品中的gDNA。所述第五组磁珠，用于识别并结合所述生物样品中的mRNA。所述第六组磁珠，用于识别并结合所述生物样品中的miRNA。所述第一组磁珠、第二 组磁珠、第三组磁珠、第四组磁珠、第五组磁珠以及第六组磁珠置于所述生物样品溶液中后，分别结合所述生物样品中的ct/cfDNA、B细胞、T细、gDNA、mDNA和miRNA，再经由所述不同孔径的通孔124流出所述套管12外，从而对所述生物样品中的ct/cfDNA、B细胞、T细、gDNA、mDNA和miRNA进行分离。
可以理解的是，每组磁珠可包括尺寸相同的多个磁珠。所述磁珠包括经过表面化学修饰的磁性纳米粒子以及结合在所述磁性纳米粒子表面的特异性分子。其中，表面化学修饰技术包括羟基化、氨基化、羧基化等处理方法，用于在所述磁性纳米粒子表面键合活性基团，使其具有良好的生物相溶性，通过与生物活性物质中的配基偶联，能够识别并结合相应的抗原、抗体或核酸。所述磁性纳米粒子的材料为铁磁性材料或顺磁性材料。顺磁性材料包括铝、镁、铂等仅较弱地被强磁体吸引的材料。铁磁性材料包括镍、钴、纯铁、铁合金等受磁体强力吸引的材料。所述特异性分子为能够结合ct/cfDNA、B细胞、T细、gDNA、mDNA或miRNA的分子。本实施例中，用于结合B细胞和T细胞的磁珠上的特异性分子为免疫抗体，用于结合ct/cfDNA、gDNA、mDNA和miRNA的磁珠上的特异性分子为寡头(Oligo)。
优选地，所述套管12包括内管122以及套设于所述内管122上的外管123。所述内管122以及外管123大致呈柱状。可以理解的是，在其他实施例中，所述内管122以及外管123还可呈球状，或其他任意形状。所述内管122以及外管123的同一端分别开设有开口，所述内管122的开口与所述外管123的开口大致平齐设置。所述内管122经由开设于所述外管123上的开口插入所述外管123中，并收容于所述外管123中。所述内管122的开口作为所述套管12的入口121。所述内管122的周壁上开设有多个第一珠孔125。所述外管123的周 壁上开设有多个第二珠孔126。所述多个第二珠孔126的位置与所述多个第一珠孔125的位置一一对应，每一第二珠孔126和相应的第一珠孔125相连通的部分形成所述通孔124。具体地，每一第二珠孔126和相应的第一珠孔125相互交叠的部分共同形成所述通孔124。本实施例中，所述第二珠孔126的尺寸和所述第一珠孔125的尺寸大致相同。
优选地，所述外管123和所述内管122能够相对转动。所述第一分离装置10进一步包括第一驱动件14。所述第一驱动件14与所述内管122和所述外管123中的一个相连接，用于驱动所述内管122和所述外管123中的一个相对另一个转动，以改变所述第二珠孔126与相应第一珠孔125的交叠面积，进而改变所述通孔124的孔径，使得所述通孔124的孔径分别与所述第一组磁珠、所述第二组磁珠、所述第三组磁珠、所述第四组磁珠、所述第五组磁珠以及第六组磁珠的尺寸相对应。需要说明的是，通孔124的孔径与磁珠的尺寸相对应指的是，所述通孔124的孔径与相应的细胞或分子结合的磁珠的尺寸大致相同，或者所述通孔124的孔径大于相应的细胞或分子结合的磁珠的尺寸但小于其他待分离的细胞或分子结合的磁珠的尺寸。具体地，所述外管123转动套设于所述内管122上；所述第一驱动件14与所述外管123相连接，用于驱动所述外管123相对所述内管122转动。可以理解的是，在其他实施例中，所述内管122可转动地设置于所述外管123中；所述第一驱动件14与所述内管122相连接，用于驱动所述内管122相对所述外管123转动。
当需要分离ct/cfDNA时，所述第一驱动件14驱动所述外管123转动，改变所述第二珠孔126与相应第一珠孔125的交叠面积，使得所述通孔124的孔径与结合有ct/cfDNA的第一组磁珠的尺寸相配合， 从而使得仅结合有ct/cfDNA的第一组磁珠由所述通孔124流出，从而分离出结合有ct/cfDNA的一磁珠。类似地，当需要分离B细胞时，所述第一驱动件14驱动所述外管123转动，改变所述通孔124的孔径，使得仅结合有B细胞的第二组磁珠能够由所述通孔124流出；当需要分离T细胞时，所述第一驱动件14驱动所述外管123转动，改变所述通孔124的孔径，使得仅结合有T细胞的第三组磁珠能够由所述通孔124流出；当需要分离gDNA时，所述第一驱动件14驱动所述外管123转动，改变所述通孔124的孔径，使得仅结合有gDNA的第四组磁珠能够由所述通孔124流出；当需要分离mRNA时，所述第一驱动件14驱动所述外管123转动，改变所述通孔124的孔径，使得仅结合有mRNA的第五组磁珠能够由所述通孔124流出；当需要分离miRNA时，所述第一驱动件14驱动所述外管123转动，改变所述通孔124的孔径，使得仅结合有miRNA的第六组磁珠能够由所述通孔124流出。可以理解的是，为使结合有ct/cfDNA、B细胞、T细胞、gDNA、mDNA以及miRNA的磁珠能够按照从小到大的顺序依次从所述通孔124流出，应依照先B细胞、T细胞、miRNA、ct/cfDNA、mRNA，最后gDNA的顺序进行分离。
可以理解的是，在其他实施例中，可以直接在所述套管12上开设具有不同孔径且可控制开闭的多个通孔124，例如设置可封盖通孔124的盖体，通过盖体的打开/关闭控制通孔124的开闭。所述多个通孔124的孔径分别与结合有ct/cfDNA、B细胞、T细胞、gDNA、mDNA或miRNA的磁珠的尺寸相配合，通过控制不同孔径通孔124的开闭，使得分别结合有ct/cfDNA、B细胞、T细胞、gDNA、mDNA以及miRNA的磁珠按照从小到大的顺序依次从所述通孔124流出。
可以理解的是，在其他实施例中，所述通孔124的数量可为一个 或两个，相应的所述第一珠孔125和第二珠孔126的数量分别为一个或两个。
优选地，所述第一分离装置进一步包括控制器16。所述控制器16与所述第一驱动件14相连接，用于控制所述第一驱动件14驱动所述外管123或内管122转动，进而控制改变所述通孔124的孔径。所述控制器16进一步控制所述通孔124的孔径变化的频率。所述控制器16控制所述通孔124的孔径由小变大，从而使得具有不同尺寸的磁珠能够按照从小到大的顺序依次单独从所述通孔124流出。
优选地，所述套管12进一步包括导管128。所述外管123的底部开设有导引孔，所述导管128与所述导引孔相连通，用于将溢出于所述外管123与所述内管122之间的缝隙中的生物样品溶液导出。
优选地，所述第一分离装置10进一步包括磁场发生器18，用于产生磁场。所述磁场发生器18产生的磁场作用于所述内管122内部，使得所述磁珠在所述磁场的作用力下吸附于所述磁场发生器18上。所述磁场发生器18用于搅拌所述生物样品溶液，使得所述多个磁珠能够由所述通孔124顺畅地流出。本实施例中，所述磁场发生器18产生的磁场为电磁场，例如所述磁场发生器18为电磁棒。可以理解的是，在其他实施例中，所述磁场发生器18产生的磁场还可为恒磁场，例如所述磁场发生器18为铷铁磁棒。
优选地，所述第一分离装置10进一步包括第二驱动件19。所述第二驱动件19与所述磁场发生器18相连接，用于驱动所述磁场发生器18进入或移出所述内管122，以及驱动所述磁场发生器18搅拌所述生物样品溶液。所述第二驱动件19进一步与所述控制器16相连接，所述控制器16控制所述第二驱动件19驱动所述磁场发生器18。
使用时，所述第一组磁珠、第二组磁珠以及第三组磁珠被注入所 述生物样品溶液中，在所述第一组磁珠结合ct/cfDNA，所述第二组磁珠结合B细胞，以及所述第三组磁珠结合T细胞后，所述第二驱动件19驱动所述磁场发生器18进入所述内管122中，所述第一组磁珠、第二组磁珠以及第三组磁珠在磁场力的作用下被吸附于所述磁场发生器18上；所述第二驱动件19驱动所述磁场发生器18移出所述内管122。所述第四组磁珠、第五组磁珠以及第六组磁珠被注入所述生物样品溶液中，在所述第四组磁珠结合gDNA，所述第五组磁珠结合mRNA，所述第六组磁珠结合miRNA后，结合有ct/cfDNA的第一组磁珠、结合有B细胞的第二组磁珠以及结合有T细胞的第三组磁珠被注入所述生物样品溶液中。当所述磁场发生器18产生的磁场为电磁场时，在所述第四组磁珠结合gDNA，所述第五组磁珠结合mRNA，所述第六组磁珠结合miRNA后，所述第二驱动件19驱动所述磁场发生器18再次进入所述内管122，所述磁场发生器18产生的电磁场被去除，例如通过切断电源的方式，使得吸附在所述磁场发生器18上的结合有ct/cfDNA的第一组磁珠、结合有B细胞的第二组磁珠以及结合有T细胞的第三组磁珠被释放于所述生物样品溶液中。当所述磁场发生器18产生的磁场为恒磁场时，在所述第二驱动件19驱动所述磁场发生器18移出所述内管122后，吸附于所述磁场发生器18上的结合有ct/cfDNA的第一组磁珠、结合有B细胞的第二组磁珠以及结合有T细胞的第三组磁珠在外力作用下脱离所述磁场发生器18；在所述第四组磁珠结合gDNA，所述第五组磁珠结合mRNA，所述第六组磁珠结合miRNA后，直接将结合有ct/cfDNA的第一组磁珠、结合有B细胞的第二组磁珠以及结合有T细胞的第三组磁珠注入生物样品溶液中。
优选地，所述分离提取装置进一步包括回收装置30，用于回收 由所述套管12流出的第一组磁珠、第二组磁珠、第三组磁珠、第四组磁珠、第五组磁珠以及第六组磁珠。具体地，所述回收装置30设置有多个通道，所述多个通道分别与所述多个通孔124一一连通，以对由所述通孔124流出的磁珠一一进行回收。可以理解的是，在其他实施例中，所述回收装置30可仅设置一个通道，经由所述多个通孔124流出的磁珠经所述通道回收于所述回收装置30中。
优选地，所述分离提取装置100进一步包括第二分离装置40，用于分离所述生物样品中的CTCs和CTM。所述第二分离装置40基于肿瘤细胞大小的分离法(ISET，isolation by size of epithelial tumor cells)进行过滤。具体地，所述第二分离装置40为滤膜，所述滤膜容置于所述套管12的内管122中，并将所述内管122的内腔分隔为上、下两部分，用于过滤所述生物样品溶液中的CTCs和CTM。在滤掉CTCs和CTM后，对所述生物样品中的溶液中的ct/cfDNA、B细胞、T细胞、ctDNA、gDNA、mDNA和miRNA进行分离。可以理解的是，在其他实施例中，所述滤膜还可以设置于所述套管12的上方或者入口处。所述滤膜可以为聚碳酸酯(Track-etched)膜、微筛(Micro Sieve)膜、微栅(TEM Grid)膜等任何可用于过滤的薄膜。本实施例中，所述滤膜的孔径为2.6～10um。CTCs一般是大于25um，核质比例很高，染色体及细胞核的形态异常的细胞。CTM是一团黏连在一起的肿瘤细胞，其尺寸较大。经过所述滤膜过滤后，尺寸较大的CTCs和CTM富集于所述滤膜上，从而分离出CTCs和CTM。分离后的CTCs和CTM可取出，用于进行染色鉴定。
优选地，在其他实施例中，所述第二分离装置40为过滤装置。所述第二分离装置40与所述第一分离装置10相连通，在所述生物样品中的CTCs和CTM经由所述第二分离装置40分离后，所述生物样 品溶液注入所述第一分离装置10以对生物样品中的ct/cfDNA、B细胞、T细胞、ctDNA、gDNA、mDNA和miRNA进行分离。所述第二分离装置40包括滤瓶以及滤膜。所述滤膜容置于所述滤瓶中，并将所述滤瓶的内腔分隔为上、下两部分，用于过滤所述生物样品溶液中的CTCs和CTM。将血液样品中的红细胞裂解后，注入所述滤瓶中，并通过所述滤膜过滤后流入所述滤瓶的底部。可以理解的是，所述滤瓶可一体成型，也可由可拆卸地两部分组装形成。
优选地，所述第二分离装置40进一步包括压力检测装置以及压力泵。所述压力泵与所述滤瓶位于所述滤膜下方的部分相连通，用于调节所述滤瓶内位于所述滤膜下方的腔体的压力，进而调节生物样品经由所述滤膜的过滤速度。所述压力检测装置用于检测所述滤瓶内位于所述滤膜下方的腔体的压力。所述压力泵依据所述压力检测装置的检测结果对所述滤瓶内位于所述滤膜下方的腔体的压力进行自动调节。
可以理解的是，在其他实施例中，所述第二分离装置40还可通过使用表面功能化的膜或管道系统吸附，从而进行CTCs分离。CTCs高表达EpCAM(epithelial cell adhesion molecule，上皮细胞黏附分子)，同时也表达Cytokeratin’s 4-6，8，10，13或19(细胞角蛋白4-6，8，10，13或19)。在膜或管道表面偶联抗EpCAM、Cytokeratin’s 4-6，8，10，13或19抗体，当生物样品流经膜或管道表面时，CTCs的表面抗原即可与相应的抗体结合，从而实现CTCs的捕获。由于CTCs不表达CD45，还可利用CD45负向筛选的方法进行CTCs分离。可以理解的是，在其他实施例中，所述第二分离装置40还可以利用迪恩涡流力学原理分选的方法进行CTCs分离。
本发明提供的分离提取装置100通过不同尺寸的磁珠结合不同 尺寸的细胞及生物分子，且通过第一驱动件14驱动外管123转动以调节所述通孔124的孔径，使得不同尺寸的磁珠能够依次单独通过所述通孔124，从而对所述生物样品中的多种细胞及生物分子同时进行分离。
请一并参阅图3，图3为本发明提供的细胞及生物分子测试系统200的模块图。所述测试系统200包括上述任一种的分离提取装置100、微流控系统210以及基因测序系统230。所述微流控系统210与所述分离提取装置100相结合，用于利用所述分离提取装置100分离出的核酸分子进行测序文库构建。所述基因测序系统230与所述微流控系统210相结合，用于对分离出的不同核酸分子进行碱基序列检测。
本发明提供的细胞及生物分子测试系统200操作中，无需人工的上样、上机等操作，实现了自动化，提高了加工效率。
以上实施方式仅用以说明本发明的技术方案而非限制，对这些实施例的多种修改对本领域的专业技术人员来说将是显而易见的，本文中所定义的一般原理可以在不脱离本发明的精神或范围的情况下，在其它实施例中实现。因此，本发明将不会被限制于本文所示的这些实施例，而是要符合与本文所公开的原理和新颖特点相一致的最宽的范围。

Claims (10)

  1. 一种细胞及生物分子分离提取装置，包括第一分离装置，其特征在于，所述第一分离装置包括：
    套管，用于盛装生物样品溶液，所述套管上开设有供生物样品溶液流出的通孔，所述通孔具有不同孔径；以及
    多组磁珠，每组磁珠用于识别并结合所述生物样品中的不同细胞或分子，其中各组磁珠之间的尺寸各不相同，所述多组磁珠分别从对应孔径的所述通孔流出。
  2. 如权利要求1所述的细胞及生物分子分离提取装置，其特征在于，所述套管包括内管以及套设于所述内管上的外管，所述内管的周壁上开设有第一珠孔，所述外管的周壁上开设有第二珠孔，所述第二珠孔和所述第一珠孔的位置一一对应，每一第二珠孔和相应的第一珠孔相连通形成所述通孔。
  3. 如权利要求2所述的细胞及生物分子分离提取装置，其特征在于，所述第一分离装置进一步包括第一驱动件，所述外管和所述内管能够相对转动，所述第一驱动件与所述外管和所述内管中的一个相连接，用于驱动所述外管和所述内管中的一个相对另一个转动，以改变所述第二珠孔与相应的第一珠孔的交叠面积，从而形成不同孔径的所述通孔。
  4. 如权利要求3所述的细胞及生物分子分离提取装置，其特征在于，所述第一分离装置进一步包括控制器，所述控制器与所述第一驱动件相连接，用于控制所述第一驱动件驱动所述外管或内管转动。
  5. 如权利要求2所述的细胞及生物分子分离提取装置，其特征在于，所述套管进一步包括导管，所述外管的底部开设有导引孔，所述导管与所述导引孔相连通。
  6. 如权利要求1所述的细胞及生物分子分离提取装置，其特征在于，所述第一分离装置进一步包括磁场发生器，用于搅拌盛装于所述套管中的生物样品溶液，所述磁场发生器发出磁场作用于所述套管内部；
    优选地，所述第一分离装置进一步包括第二驱动件，所述第二驱动件与所述磁场发生器相连接，用于驱动所述磁场发生器进入及移出所述套管。
  7. 如权利要求6所述的细胞及生物分子分离提取装置，其特征在于，所述多组磁珠包括：
    第一组磁珠，用于识别并结合所述生物样品中的ct/cfDNA；
    第二组磁珠，用于识别并结合所述生物样品中的B细胞；
    第三组磁珠，用于识别并结合所述生物样品中的T细胞；
    第四组磁珠，用于识别并结合所述生物样品中的gDNA；
    第五组磁珠，用于识别并结合所述生物样品中的mRNA；以及
    第六组磁珠，用于识别并结合所述生物样品中的miRNA；
    优选地，在所述第一组磁珠结合ct/cfDNA，所述第二组磁珠结合B细胞，以及所述第三组磁珠结合T细胞后，所述第二驱动件驱动所述磁场发生器进入所述套管；在所述磁场发生器吸附所述第一组磁珠、第二组磁珠以及第三组磁珠后，所述第二驱动件驱动所述磁场发生器移出所述套管；在所述第四组磁珠结合gDNA，所述第五组磁珠结合mRNA，所述第六组磁珠结合miRNA后，结合有ct/cfDNA的第一组磁珠、结合有B细胞的第二组磁珠以及结合有T细胞的第三组磁珠被注入所述生物样品溶液中；
    优选地，所述磁场发生器产生的磁场为电磁场，在所述第四组磁珠结合gDNA，所述第五组磁珠结合mRNA，所述第六组磁珠结合 miRNA后，所述第二驱动件驱动所述磁场发生器再次进入所述套管，所述磁场发生器产生的电磁场被去除，使得吸附在所述磁场发生器上的第一组磁珠、第二组磁珠以及第三组磁珠被释放于所述套管中。
  8. 如权利要求1所述的细胞及生物分子分离提取装置，其特征在于，进一步包括回收装置，用于回收由所述套管流出的第一组磁珠、第二组磁珠、第三组磁珠、第四组磁珠、第五组磁珠以及第六组磁珠；
    优选地，所述回收装置设置有多个通道，所述多个通道分别与所述多个通孔一一连通。
  9. 如权利要求1所述的细胞及生物分子分离提取装置，其特征在于，进一步包括第二分离装置，用于分离所述生物样品中的CTCs和CTM；
    优选地，所述第二分离装置为滤膜。
  10. 一种细胞及生物分子测试系统，其特征在于，包括权利要求1-9中任一项所述的细胞及生物分子分离提取装置、微流控系统以及基因测序系统，所述微流控系统与所述细胞及生物分子分离提取装置相结合，所述基因测序系统与所述微流控系统相结合。
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