WO2019174468A1 - 一种核酸提取纯化正压装置 - Google Patents

一种核酸提取纯化正压装置 Download PDF

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WO2019174468A1
WO2019174468A1 PCT/CN2019/076453 CN2019076453W WO2019174468A1 WO 2019174468 A1 WO2019174468 A1 WO 2019174468A1 CN 2019076453 W CN2019076453 W CN 2019076453W WO 2019174468 A1 WO2019174468 A1 WO 2019174468A1
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nucleic acid
positive pressure
column
purification
acid extraction
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PCT/CN2019/076453
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English (en)
French (fr)
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易初丽
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武汉医蒂生物科技有限公司
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    • 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
    • C12M1/00—Apparatus for enzymology or microbiology
    • 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
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/12—Apparatus for enzymology or microbiology with sterilisation, filtration or dialysis means
    • 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
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/26—Inoculator or sampler
    • 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
    • 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

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  • the invention relates to the field of experimental devices, in particular to a nucleic acid extraction and purification positive pressure device.
  • nucleic acid cleaning and nucleic acid elution processes There are many methods for extracting or purifying nucleic acids from biological tissues, but the more common method at present is the column method, which requires a centrifuge or a centrifuge plus a negative pressure pump to complete most of its processes such as debris filtration. , nucleic acid cleaning and nucleic acid elution processes.
  • the solution cannot pass the resistance of the membrane at atmospheric pressure, so it is necessary to use centrifugal force or negative The pressure causes the solution of the extracted or purified nucleic acid to flow out through the resistance of the membrane.
  • the nucleic acid filter column or the adsorption column containing the nucleic acid solution to be extracted is required to be placed in a centrifuge tube, and then centrifuged for several minutes in the centrifuge, and then they are taken out from the centrifuge and the solution in the centrifuge tube is poured out.
  • the nucleic acid extraction or purification process needs to repeat the above process 8-10 times.
  • the centrifugation method requires the nucleic acid filtration column or the adsorption column to be centrifuged in the centrifuge repeatedly, which is troublesome and time consuming.
  • the centrifugal method plus the negative pressure method simplifies the operation and reduces the time, but the efficiency is still Very low, the reason why the negative pressure method can not completely replace the centrifugal method is because the suction pipe of the negative pressure pump cannot be directly connected to the water outlet of the filter column or the adsorption column, but must be connected with a negative pressure tank (the solution tray is inside the tank).
  • the solution tray is arranged below the liquid outlet of the filter column or the adsorption column, and the negative pressure tank has a certain blocking effect on the negative pressure of the negative pressure pump, so that the negative pressure pump acts on the negative pressure of the filter column or the adsorption column. It is obviously reduced. Furthermore, the negative pressure pump that can be purchased to the maximum can not exceed the negative pressure of -100 kPa, so some steps of nucleic acid extraction or purification must be completed with a centrifugal force of a large force.
  • the embodiments of the present invention provide a nucleic acid extraction and purification positive pressure device.
  • the technical solution is as follows:
  • the embodiment of the invention provides a nucleic acid extraction and purification positive pressure device
  • the nucleic acid extraction and purification positive pressure device comprises: a sealing plugging column, a connecting tube, a positive pressure provider, and a sequentially connected filter column and an adsorption column, the filter column
  • the filter column For the round table structure, the top of the filter column is provided with a filter column inlet, the bottom of the filter column is provided with a filter column outlet, the filter column inlet and the filter column outlet
  • a filter membrane is disposed on an inner wall of the filter column, the adsorption column is a round table structure, a top of the adsorption column is provided with an adsorption column inlet, and a bottom of the adsorption column is provided with an adsorption column.
  • An adsorption film is disposed on an inner wall of the adsorption column between the liquid inlet of the adsorption column and the liquid outlet of the adsorption column, and the bottom of the filter column may be clamped on the liquid inlet of the adsorption column;
  • the sealing plug column is a round table structure respectively sealingly matched with the filter column inlet port and the adsorption column inlet port, and the sealing plug column is open to communicate with the filter column or the adsorption column Connected hole
  • a communication tube sleeve is disposed on the communication hole, and one end of the communication tube is sleeved on the communication tube sleeve, and the other end of the communication tube is in sealing communication with an air outlet of the positive pressure provider.
  • the sealing plug is an elastic sealing plug.
  • sealing plug is a plug of silica gel.
  • the positive pressure provider provides a pressure of 100-500 kPa.
  • the positive pressure provider provides a pressure of 250 kPa.
  • the positive pressure provider is an air compression pump, a piston air pump, a diaphragm pump or a liquid nitrogen bottle.
  • the communication pipe is a rubber hose.
  • the nucleic acid extraction and purification positive pressure device further comprises a test tube rack, and the test tube rack is provided with a test tube hole matched with the adsorption column, and the adsorption column is installed in the test tube hole.
  • the nucleic acid extraction and purification positive pressure device further includes a waste liquid tray and a collection tube, wherein the waste liquid tray and the collection tube are both located in the test tube rack and are respectively disposed under the two test tube holes respectively.
  • the technical solution provided by the embodiments of the present invention has the beneficial effects that the nucleic acid extraction and purification positive pressure device uses a positive pressure provider to provide positive pressure on the filter column and the adsorption column, and the positive pressure provider can generate higher pressure. So that it can replace the centrifuge or the combination of the centrifuge and the negative pressure pump to complete the whole process of nucleic acid extraction and purification, so that the operation of nucleic acid extraction and purification is compared with the use of a centrifuge or a combination of a centrifuge and a negative pressure pump.
  • FIG. 1 is a schematic view showing the structure of a nucleic acid extraction and purification positive pressure device equipped with a sealing plug column according to an embodiment of the present invention
  • FIG. 2 is a schematic structural view of a sealing plugging column and an adsorption column provided by an embodiment of the present invention.
  • the embodiment of the invention provides a nucleic acid extraction and purification positive pressure device.
  • the nucleic acid extraction and purification positive pressure device comprises: a sealing plug 3, a communication tube 4, a positive pressure provider 5 and a filter column connected in sequence. 1 and adsorption column 2.
  • the filter column 1 has a round table structure, the top of the filter column 1 is provided with a filter column inlet port 1a, and the bottom of the filter column 1 is provided with a filter column outlet port 1b, the filter column inlet port 1a and the filter column outlet port 1b.
  • a filter membrane (not shown) is disposed on the inner wall of the filter column 1 , the adsorption column 2 is a round table structure, the top of the adsorption column 2 is provided with a suction column inlet 2a, and the bottom of the adsorption column 2 is provided with An adsorption film (not shown) is disposed on the inner wall of the adsorption column 2b between the adsorption column inlet port 2a and the adsorption column inlet port 2b, and the bottom of the filter column 1 is attached to the adsorption column. On the liquid inlet 2a.
  • the sealing plug 3 is a round table structure which is respectively sealed with the filter column inlet port 1a and the adsorption column inlet port 2a, and the sealing plug column 3 is provided with a communication hole 3a communicating with the filter column 1 or the adsorption column 2;
  • a communication tube sleeve 8 is disposed on the hole 3a, and one end of the communication tube 4 is sealingly mounted on the communication tube sleeve 8, and the other end of the communication tube 4 is in sealing communication with the air outlet of the positive pressure provider 5.
  • the filter membrane can be overlaid above the filter column outlet 1b.
  • the adsorption film may be covered above the adsorption column outlet 2b.
  • the sealing plug 3 can be an elastic sealing plug.
  • the elastic sealing plug can be a silicone sealing plug or a rubber sealing plug, which makes the sealing plug 3 have good elasticity, and is used for further improving the sealing effect of the sealing plug 3 .
  • the sealing plug 3 can be a silicone plug.
  • the silicone plug has low cost and good flexibility, and can achieve a good sealing effect.
  • the pressure provided by the positive pressure supply device 5 may be 100 to 500 kPa.
  • the positive pressure provider 5 provides a pressure greater than 250 kPa, the nucleic acid-containing solution can be quickly passed through a filtration membrane or adsorption membrane having a pore size of 0.2 to 25 ⁇ m.
  • the pressure provided by the positive pressure provider 5 may be 110 to 250 kPa. At this pressure range, the solution can be smoothly passed through the filtration column 1 or the adsorption column 2.
  • the positive pressure provider 5 provides a pressure of 250 kPa.
  • the conventional filter column 1 and the adsorption column 2 can be fitted with a positive pressure provider 5 having a pressure of 250 kPa. If the pore size of the filtration membrane or the adsorption membrane is small, and the volume of the solution to be extracted and purified is large, the pressure provided by the positive pressure provider 5 may be increased, for example, greater than 250 kPa, and if the solution contains impurities such as blood clots, The pressure provided by the positive pressure provider 5 is appropriately increased to 400 to 500 kPa.
  • the positive pressure provider 5 may be an air compression pump, a piston air pump, a diaphragm pump, or a liquid nitrogen bottle.
  • the positive pressure provider 5 may be a miniature piston air pump, and the volume of the miniature piston air pump is about 40 mm ⁇ 58 mm ⁇ 120 mm (width ⁇ height ⁇ length), the weight is about 483 g, and the flow rate is 7 ⁇ . 10L/min, the cost is about 400 yuan. Due to the low cost, small size and light weight of the miniature piston air pump, the cost, volume and weight of the nucleic acid extraction and purification positive pressure device are far lower than the combination of a centrifuge or a centrifuge and a negative pressure pump.
  • the positive pressure provider 5 can be simultaneously performed for a plurality of filter columns 1 or a plurality of adsorption columns 2.
  • positive pressure can be increased.
  • the power and flow of the provider 5 can also be equipped with a gas storage tank for the air compression pump or the piston air pump.
  • the communication tube 4 may be a rubber hose.
  • the rubber hose has good toughness and is not easy to be broken, and the rubber hose can arbitrarily arrange the position of the positive pressure provider 5 without being limited by space.
  • the nucleic acid extraction and purification positive pressure device may further include a test tube rack 9 on which a test tube hole 9a that cooperates with the adsorption column 2 is opened, and the adsorption column 2 is installed in the test tube hole 9a.
  • the test tube rack 9 may be provided with two layers, the first layer is provided with a plurality of test tube holes 9a and used for mounting the adsorption column 2, and the second layer is for placing the collection tube 10 and the waste liquid tray 11 and the like.
  • the nucleic acid extraction and purification positive pressure device may further include a collection tube 10 and a waste liquid tray 11, and the collection tube 10 and the waste liquid tray 11 are both located in the second layer of the test tube rack 9 and are respectively arranged correspondingly in the two test tube holes 9a.
  • the waste tray 11 is used to collect the waste liquid
  • the collection tube 10 is used to collect the nucleic acid solution.
  • the respective components are connected in position as shown in Fig. 1: the adsorption column 2 is inserted into the test tube hole 9a of the test tube rack 9; the filter column 1 is snapped onto the adsorption column inlet 2a of the adsorption column 2 to filter The column liquid outlet 1b is aligned with the adsorption column inlet port 2a; the communication tube sleeve 8 is inserted into the communication hole 3a from the bottom of the sealing plug column 3, and the communication tube 4 connected to the positive pressure provider 5 is tightly inserted into the communication tube sleeve. Inside the barrel 8.
  • the nucleic acid solution to be extracted or purified is injected into the filter column 1; the sealing plug 3 is sealed and installed on the filter column outlet port 1b, and the high pressure gas is introduced into the filter column 1 through the communication tube 4 to make the high pressure The gas acts on the upper surface of the solution, and the solution is rapidly passed through the filter membrane by the high pressure.
  • the impurities are filtered out, and the nucleic acid flows out of the filter column outlet port 1b to the adsorption column 2 as the solution passes through the filter membrane.
  • the sealing plug 3 is taken out from the filter column inlet 1a and the filter column 1 is discarded;
  • Adsorbing nucleic acid as shown in FIG. 2, the sealing plug 3 is sealed and installed on the adsorption column inlet 2a, and a high-pressure gas is introduced into the adsorption column 2 through the communication pipe 4, so that the high-pressure gas acts on the upper surface of the solution, and the solution The high-pressure action rapidly passes through the adsorption membrane. After the solution passes through the adsorption membrane, the nucleic acid is adsorbed on the adsorption membrane, and the other solution flows out from the adsorption column outlet 2b and flows into the waste liquid tray 11;
  • the sealing plug 3 is removed from the adsorption column inlet 2a, and the cleaning liquid is injected into the adsorption column 2, and then the sealing plug 3 is sealed and installed on the adsorption column inlet 2a, through the communication tube 4
  • the high-pressure gas is introduced into the adsorption column 2, and the cleaning liquid rapidly passes through the adsorption membrane to clean the nucleic acid in the membrane under the action of the high-pressure gas, and then flows from the adsorption column outlet port 2b to the waste liquid tray 11 below it, repeating the cleaning process.
  • the nucleic acid in the adsorption membrane is thoroughly washed 2-3 times, and the nucleic acid is retained on the adsorption membrane;
  • the sealing plug 3 is removed from the adsorption column inlet 2a, and the eluent is injected into the adsorption column 2, and then the sealing plug 3 is sealed and installed on the adsorption column inlet 2a, and at the same time,
  • the waste liquid tray 11 is removed and the collection tube 10 is moved to the lower side of the nucleic acid adsorption column outlet port 2b, and a high-pressure gas is introduced into the adsorption column 2 through the communication tube 4, so that the nucleic acid eluent is rapidly passed by the high pressure of the high pressure gas.
  • the adsorption film allows the nucleic acid adsorbed on the adsorption film to be quickly eluted, and is discharged from the adsorption column outlet 2b to the collection tube 10 to complete the extraction and purification process of the nucleic acid.
  • the embodiment of the invention provides a nucleic acid extraction and purification positive pressure device, which uses a positive pressure provider to provide positive pressure on the filter column and the adsorption column, and the positive pressure provider can generate high pressure. So that it can replace the centrifuge or the combination of the centrifuge and the negative pressure pump to complete the whole process of nucleic acid extraction and purification, and make the operation of nucleic acid extraction and purification relative to the use of a centrifuge or a combination of a centrifuge and a negative pressure pump. It is much simpler and the pressure generated by the positive pressure provider is large, which makes the nucleic acid extraction purification process using the nucleic acid extraction and purification positive pressure device faster than the centrifuge 10 times higher than the centrifuge plus negative pressure pump. Five times this, the efficiency of nucleic acid extraction or purification is greatly improved. At the same time, the rounded table type sealing plug can adapt to a variety of different specifications of the filter column or adsorption column, so that the sealing block has better adaptability.

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Abstract

一种核酸提取纯化正压装置，包括：密封堵柱、连通管、正压提供器以及顺序连接的过滤柱和吸附柱。该核酸提取纯化正压装置能够快速高效提取纯化核酸。

Description

一种核酸提取纯化正压装置
本申请要求于2018年03月13日提交中国专利局、申请号为201810206062.6、发明名称为“一种核酸提取纯化正压装置”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
本发明涉及实验装置领域，特别涉及一种核酸提取纯化正压装置。
背景技术
从生物组织里提取或纯化核酸的方法有多种，但目前较常用的方法是柱式法，这种方法需要借用离心机或离心机加负压泵才能完成其大部分的过程如杂物过滤、核酸清洗和核酸洗脱的过程。
由于常用的核酸提取过滤柱子和吸附柱子内的过滤膜和吸附膜的孔径都很小，一般在0.7～5.0微米左右，大气压下溶液是无法通过这种膜的阻力的，故需要借助离心力或负压力使提取或纯化核酸的溶液通过膜的阻力流出。
利用离心力时需要将装有待提取核酸溶液的核酸过滤柱或吸附柱外套装一个离心管后放入离心机内离心数分钟，然后又要将它们从离心机内取出倒掉离心管内的溶液，整个核酸提取或纯化过程需要重复上述过程8-10次左右。
为了简化上述使用离心机的操作麻烦和时间的消耗，通常在核酸提取或纯化时的过程中有些不需要较大力量的步骤便利用负压力代替离心力：将过滤柱或吸附柱插入到一个负压箱上，负压箱与一负压泵相连，通过负压力时柱子内的溶液流出。
在实现本发明的过程中，发明人发现现有技术至少存在以下问题：
离心法需要将核酸过滤柱或吸附柱反复多次地放入离心机内离心，这样操作较麻烦且耗时较长；离心法加负压法虽然简化操作麻烦和减少了时间，但是其效率仍然很低下，负压法不能完全替代离心法的原因是因为负 压泵的吸气管子不能直接与过滤柱或吸附柱的出水口相连，而必须需与一负压箱(箱内有溶液盘)连接，溶液盘布置在过滤柱或吸附柱的出液口的下方，负压箱对负压泵的负压力有一定的阻隔作用，这样使得负压泵作用在过滤柱或吸附柱上的负压力就明显地减小了，再者，目前能购达到最大的实验用的负压泵不能超过-100kPa的负压，故核酸提取或纯化的有些步骤必须要用较大力量的离心力来完成。
发明内容
为了解决现有技术中需要利用离心机和负压泵所带来的操作上的麻烦和时间的浪费，本发明实施例提供了一种核酸提取纯化正压装置。所述技术方案如下：
本发明实施例提供了一种核酸提取纯化正压装置，所述核酸提取纯化正压装置包括：密封堵柱、连通管、正压提供器以及顺序连接的过滤柱和吸附柱，所述过滤柱为倒圆台型结构，所述过滤柱的顶部设置有过滤柱进液口，所述过滤柱的底部设置有过滤柱出液口，所述过滤柱进液口与所述过滤柱出液口之间的所述过滤柱的内壁上设置有过滤膜，所述吸附柱为倒圆台型结构，所述吸附柱的顶部设置有吸附柱进液口，所述吸附柱的底部设置有吸附柱出液口，所述吸附柱进液口与所述吸附柱出液口之间的所述吸附柱的内壁上设置有吸附膜，所述过滤柱的底部可卡装在所述吸附柱进液口上；
所述密封堵柱为分别与所述过滤柱进液口和所述吸附柱进液口密封配合的倒圆台型结构，所述密封堵柱上开设有与所述过滤柱或所述吸附柱相通的连通孔；
所述连通孔上设置有连通管套筒，所述连通管的一端套装在所述连通管套筒上，所述连通管的另一端与所述正压提供器的出气口密封连通。
具体地，所述密封堵柱为弹性密封堵柱。
进一步地，所述密封堵柱为硅胶堵柱。
进一步地，所述正压提供器提供的压强为100-500kPa。
优选地，所述正压提供器提供的压强为250kPa。
更进一步地，所述正压提供器为空气压缩泵、活塞气泵、隔膜泵或液 氮瓶。
具体地，所述连通管为橡胶软管。
具体地，所述核酸提取纯化正压装置还包括试管架，所述试管架上开设有与所述吸附柱相配合的试管孔，所述吸附柱安装在所述试管孔内。
具体地，所述核酸提取纯化正压装置还包括废液盘和收集管，所述废液盘和所述收集管均位于所述试管架内并分别对应布置在两个所述试管孔的下方。
本发明实施例提供的技术方案带来的有益效果是：该核酸提取纯化正压装置采用正压提供器作用在过滤柱和吸附柱上提供正压，由于正压提供器可产生较高的压力，使得其能取代离心机或离心机与负压泵的组合完成核酸的提取纯化的全过程，这样使得核酸提取纯化的操作过程相对于采用离心机或用离心机与负压泵的组合的方式要简单得多，且正压提供器所产生的压力大，这使得采用该核酸提取纯化正压装置进行核酸提取纯化的速度高于用离心机的10倍，高于用离心机加负压泵的5倍，这样使得核酸提取或纯化的效率得到大大地提高。
附图说明
为了更清楚地说明本发明实施例中的技术方案，下面将对实施例描述中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图仅仅是本发明的一些实施例，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他的附图。
图1是本发明实施例提供的核酸提取纯化正压装置装配有密封堵柱的结构示意图；
图2是本发明实施例提供的密封堵柱配合吸附柱的结构示意图。
具体实施方式
为使本发明的目的、技术方案和优点更加清楚，下面将结合附图对本 发明实施方式作进一步地详细描述。
实施例一
本发明实施例提供了一种核酸提取纯化正压装置，如图1所示，该核酸提取纯化正压装置包括：密封堵柱3、连通管4、正压提供器5以及顺序连接的过滤柱1和吸附柱2。过滤柱1为倒圆台型结构，过滤柱1的顶部设置有过滤柱进液口1a，过滤柱1的底部设置有过滤柱出液口1b，过滤柱进液口1a与过滤柱出液口1b之间的过滤柱1的内壁上设置有过滤膜(图中未示)，吸附柱2为倒圆台型结构，吸附柱2的顶部设置有吸附柱进液口2a，吸附柱2的底部设置有吸附柱出液口2b，吸附柱进液口2a与吸附柱出液口2b之间的吸附柱2的内壁上设置有吸附膜(图中未示)，过滤柱1的底部卡装在吸附柱进液口2a上。
密封堵柱3为分别与过滤柱进液口1a和吸附柱进液口2a密封配合的倒圆台型结构，密封堵柱3上开设有与过滤柱1或吸附柱2相通的连通孔3a；连通孔3a上设置有连通管套筒8，连通管4的一端密封安装在连通管套筒8上，连通管4的另一端与正压提供器5的出气口密封连通。
使用时，过滤膜可以覆盖在过滤柱出液口1b上方。吸附膜可以覆盖在吸附柱出液口2b上方。
具体地，密封堵柱3可以为弹性密封堵柱。弹性密封堵柱可以为硅胶密封堵柱或橡胶密封堵柱，这使得密封堵柱3具有良好的弹性，用于进一步提高密封堵柱3的密封效果。
进一步地，密封堵柱3可以为硅胶堵头。硅胶堵头成本低、柔韧性好，可实现良好的密封效果。
具体地，正压提供装器5提供的压强可以为100～500kPa。当正压提供器5提供压强大于250kPa时，能够使含有核酸的溶液快速地穿过孔径在0.2～25微米的过滤膜或吸附膜。
进一步地，正压提供器5提供的压强可以为110～250kPa。在该压强范围下能够使溶液顺利通过过滤柱1或吸附柱2。
优选地，正压提供器5提供的压强可以为250kPa。常规的过滤柱1和吸附柱2可以配合的压强为250kPa的正压提供器5。若过滤膜或吸附膜的孔径较小，且需要提取纯化的溶液体积较多时，可以增加正压提供器5提 供的压强，如大于250kPa，此外，若溶液中含有血凝块等杂质时也可以适当增加正压提供器5提供的压强至400～500kpa。
具体地，该正压提供器5可以为空气压缩泵、活塞气泵、隔膜泵或液氮瓶。在本实施例中，正压提供器5可以为微型的活塞气泵，且该微型的活塞气泵的体积约为40mm×58mm×120mm(宽×高×长)，重量约为483g，流量为7～10L/min，成本为400元左右。由于微型的活塞气泵成本低、体积小且重量轻，这使得该核酸提取纯化正压装置的成本、体积和重量都远远低于离心机或离心机与负压泵的组合。
另外，正压提供器5可以针对多个过滤柱1或多个吸附柱2同时进行。当需要提取纯化的核酸的数量较多时，例如每次需要提取纯化的核酸数量多于十个时，或者，同时为多个过滤柱1或多个吸附柱2提供正压时，可以增加正压提供器5的功率和流量，还可以为空气压缩泵或活塞气泵配备储气罐。
具体地，连通管4可以为橡胶软管。橡胶软管韧性好，不易折断，橡胶软管可以使正压提供器5的位置任意布置，而不受空间限制。
如图1所示，具体地，该核酸提取纯化正压装置还可以包括试管架9，试管架9上开设有与吸附柱2相配合的试管孔9a，吸附柱2安装在试管孔9a内。在实现时，试管架9可以设置2层，第一层开设有多个试管孔9a并用于安装吸附柱2，第二层用于放置收集管10和废液盘11等用。
具体地，该核酸提取纯化正压装置还可以包括收集管10和废液盘11，收集管10和废液盘11均位于试管架9的第二层内并分别对应布置在两个试管孔9a的下方，废液盘11用于收集废液，收集管10用于收集核酸溶液。
下面简单介绍一下本发明实施例提供的核酸提取纯化正压装置的工作原理，具体如下：
参见图1，首先将各个部件如图1连接到位：将吸附柱2插入到试管架9的试管孔9a内；将过滤柱1卡装在吸附柱2的吸附柱进液口2a上，使过滤柱出液口1b对准吸附柱进液口2a；将连通管套筒8从密封堵柱3的底部插入连通孔3a内并将与正压提供器5相连的连通管4紧密插入连通管套筒8内。
过滤杂物：向过滤柱1内注入待提取或纯化的核酸溶液；将密封堵柱3密封安装在过滤柱出液口1b上，通过连通管4向过滤柱1内通入高压气体，使高压气体作用在溶液的上表面，溶液受到高压作用快速通过过滤膜，溶液经过过滤膜时杂物被过滤掉，而核酸则随溶液通过过滤膜后由过滤柱出液口1b流出至吸附柱2内，此时过滤柱1的工作完成。然后将密封堵柱3从过滤柱进液口1a处取出并扔弃过滤柱1；
吸附核酸：如图2所示，将密封堵柱3密封安装在吸附柱进液口2a上，通过连通管4向吸附柱2内通入高压气体，使高压气体作用在溶液的上表面，溶液受到高压作用快速通过吸附膜，溶液经过吸附膜后，核酸被吸附在吸附膜上，其它溶液由吸附柱出液口2b流出，并流入废液盘11中；
清洗核酸：将密封堵柱3由吸附柱进液口2a上移开，向吸附柱2内注入清洗液后再次将密封堵柱3密封安装在吸附柱进液口2a上，通过连通管4向吸附柱2内通入高压气体，清洗液在高压气体的作用下快速通过吸附膜清洗膜内的核酸后由吸附柱出液口2b流到其下方的废液盘11内，重复这种清洗过程2-3次彻底清洗吸附膜内的核酸，核酸滞留在吸附膜上；
洗脱核酸：将密封堵柱3由吸附柱进液口2a上移开，向吸附柱2内注入洗脱液后再次将密封堵柱3密封安装在吸附柱进液口2a上，同时，将废液盘11移开并将收集管10移动到核酸吸附柱出液口2b的下面，通过连通管4向吸附柱2内通入高压气体，使核酸洗脱液受到高压气体的高压作用快速通过吸附膜，使吸附在吸附膜上的核酸能够被快速洗脱下来，并由吸附柱出液口2b流出至收集管10中，完成核酸的提取纯化过程。
本发明实施例提供了一种核酸提取纯化正压装置，该核酸提取纯化正压装置采用正压提供器作用在过滤柱和吸附柱上提供正压，由于正压提供器可产生较高的压力，使得其能取代离心机或离心机与负压泵的组合完成核酸的提取纯化的全过程，并使得核酸提取纯化的操作过程相对于采用离心机或用离心机与负压泵的组合的方式要简单得多，且正压提供器所产生的压力大，这使得采用该核酸提取纯化正压装置进行核酸提取纯化的速度高于用离心机的10倍，高于用离心机加负压泵的5倍，这样使得核酸提取或纯化的效率得到大大地提高。同时，倒圆台型的密封堵柱能够适应多种 不同规格的过滤柱或吸附柱，从而使密封堵柱具有更好的适应性。
以上所述仅为本发明的较佳实施例，并不用以限制本发明，凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在发明的保护范围之内。
Figure PCTCN2019076453-appb-000001

Claims (2)

  1. 根据权利要求1所述的核酸提取纯化正压装置，其特征在于，所述核酸提取纯化正压装置还包括试管架，所述试管架上开设有与所述吸附柱相配合的试管孔，所述吸附柱安装在所述试管孔内。
  2. 根据权利要求7所述的核酸提取纯化正压装置，其特征在于，所述核酸提取纯化正压装置还包括废液盘和收集管，所述废液盘和所述收集管均位于所述试管架内并分别对应布置在两个所述试管孔的下方。
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