WO2024222956A1 - 全自动一体化核酸提取设备 - Google Patents

全自动一体化核酸提取设备 Download PDF

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WO2024222956A1
WO2024222956A1 PCT/CN2024/090407 CN2024090407W WO2024222956A1 WO 2024222956 A1 WO2024222956 A1 WO 2024222956A1 CN 2024090407 W CN2024090407 W CN 2024090407W WO 2024222956 A1 WO2024222956 A1 WO 2024222956A1
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hole
negative pressure
nucleic acid
acid extraction
reaction chamber
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PCT/CN2024/090407
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French (fr)
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糜军
张洁莹
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糜军
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Publication of WO2024222956A1 publication Critical patent/WO2024222956A1/zh

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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
    • C12M1/42—Apparatus for the treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
    • 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

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  • the present application relates to a nucleic acid extractor, and in particular to a fully automatic integrated nucleic acid extraction device.
  • the detection methods for pathogenic microorganisms mainly include microscopy, culture, immunological testing, and nucleic acid amplification testing.
  • the microscopy method is low-cost, simple, and convenient, but the positive rate is low and it is easy to miss detection; the results of the culture method are highly reliable, but the cycle is long and the cost is high; the operation of immunological testing is relatively simple, but there is a window period, so it is easy to miss detection.
  • nucleic acid testing are equivalent to those of the culture method, but there are also the following shortcomings: 1 Sample preparation (nucleic acid extraction) and nucleic acid amplification/detection are carried out separately, the operation steps are cumbersome, and professional personnel are required to perform, which increases the operation error; 2 Open nucleic acid extraction is also prone to cross-contamination of samples, resulting in false positive test results; 3 Although the operation cycle of nucleic acid testing is shorter than that of pathogenic microorganism culture, it is also difficult to meet the requirements of bedside testing (POCT). How to quickly amplify nucleic acids, shorten detection time, and reduce patient waiting time has become a top priority.
  • POCT bedside testing
  • the purpose of the present application is to provide a fully automatic integrated nucleic acid extraction device to integrate the device and reduce cross contamination.
  • the present application discloses a fully automatic integrated nucleic acid extraction device, which comprises: a shell, a sample injection hole, a first negative pressure hole, a second negative pressure hole, a magnetic rod insertion hole, a sampling hole, a first negative pressure pipeline and a second negative pressure pipeline;
  • the shell is square, the sample injection hole, the magnetic rod insertion hole, the first negative pressure hole and the second negative pressure hole are arranged on the top of the shell, the sampling hole is arranged on the side of the shell,
  • the shell comprises a waste chamber and a reaction chamber, the waste chamber and the reaction chamber are separated by a partition, a one-way valve is arranged inside the reaction chamber, the reaction chamber is separated into an upper reaction chamber and a lower reaction chamber, the upper reaction chamber and the first negative pressure pipeline are connected to the waste chamber, and the second negative pressure pipeline is connected to the lower reaction chamber through the second negative pressure hole.
  • the sample injection hole includes a sample addition hole and a solution injection hole.
  • the solution injection hole is connected to a multi-directional pump, and the nucleic acid extraction of pathogenic microorganisms is completed by controlling the injection of different liquids.
  • magnetic beads and a magnetic sleeve are arranged inside the reaction chamber.
  • the sampling hole is connected to the lower reaction chamber.
  • the first negative pressure pipeline and the second negative pressure pipeline are connected to a negative pressure pump.
  • the device is equipped with a syringe pump, and the syringe pump is connected to the solution injection hole.
  • the device is equipped with an ultrasonic generator and a heating module, and the ultrasonic generator and the heating module are closely attached to the side and back of the nucleic acid extraction device.
  • the magnetic rod insertion hole is arranged between the sample addition hole and the solution injection hole.
  • a sampling rod is provided at the sampling hole.
  • feature A+B+C is disclosed
  • feature A+B+D+E is disclosed
  • features C and D are equivalent technical means that play the same role.
  • Feature E can be combined with feature C technically. Then, the solution of A+B+C+D should not be deemed to have been recorded because it is technically infeasible, and the solution of A+B+C+E should be deemed to have been recorded.
  • FIG1 is a schematic diagram of the overall structure of a nucleic acid extraction box according to an embodiment of the present invention.
  • FIG2 is a side cross-sectional view of a nucleic acid extraction box according to an embodiment of the present invention.
  • FIG3 is a side view of a nucleic acid extraction box according to an embodiment of the present invention.
  • FIG. 4 is a side cross-sectional view of a nucleic acid extraction device from another angle according to an embodiment of the present invention.
  • the device has no cross contamination during the detection process and has high nucleic acid extraction efficiency; it has a simple structure and relatively low cost, which is convenient for integration with nucleic acid amplification equipment; it is simple to operate, not prone to errors, has good repeatability of results, and has extremely low requirements for operators and hardware facilities used, and has good applicability, which can simultaneously meet the sample detection needs of clinical outpatient clinics and primary medical institutions; it saves the most time, and the entire nucleic acid extraction operation time does not exceed 10 minutes.
  • FIGS. 1 to 4 An embodiment of the present invention is shown in FIGS. 1 to 4 , which includes a housing, a sample injection hole, a first negative pressure hole 4, a second negative pressure hole 5, a magnetic rod insertion hole 2, a sampling hole 6, a first negative pressure pipeline 9 and a second negative pressure pipeline 10; wherein:
  • the shell is square, the sample injection hole 6, the magnetic rod insertion hole 2, the first negative pressure hole 4 and the second negative pressure hole 5 are arranged on the top of the shell, and the sampling hole 6 is arranged on the side of the shell.
  • the sample injection hole includes a sample addition hole 1 and a solution injection hole 3.
  • the solution injection hole 3 is connected to a multi-directional pump to control the injection of different liquids to complete the nucleic acid extraction of pathogenic microorganisms.
  • the interior of the shell includes a waste chamber 11 and a reaction chamber 7.
  • magnetic beads are arranged inside the reaction chamber 7.
  • the waste chamber 11 and the reaction chamber 7 are separated by a partition.
  • a one-way valve 8 is arranged inside the reaction chamber 7 to separate the reaction chamber 7 into an upper reaction chamber and a lower reaction chamber.
  • the upper reaction chamber and the first negative pressure pipe 9 are connected to the waste chamber 11, and the second negative pressure pipe 10 is connected to the lower reaction chamber through the second negative pressure hole 5.
  • the first negative pressure pipe 9 and the second negative pressure pipe 10 are connected to a negative pressure pump connect.
  • the nucleic acid extraction device is further configured with an ultrasonic generator and a heating module, and the ultrasonic generator and the heating module are closely attached to the side and back of the nucleic acid extraction device to provide ultrasonic vibration and heating during use.
  • a sampling rod is provided at the sampling hole 6.
  • the nucleic acid extraction device further comprises a magnetic rod 12, which is inserted into the reaction chamber 7 through the magnetic rod insertion hole 2 for nucleic acid extraction.
  • the magnetic rod is provided with a magnetic sleeve, which is embedded in the reaction chamber.
  • the process of this embodiment during use is as follows: first, a certain amount of sputum or mouthwash containing pathogenic microorganisms is added through the sample addition hole, the sample is mixed with the pre-filled lysis solution, and pathogenic microorganisms such as viruses and bacteria are lysed by ultrasound and heating methods to release nucleic acid substances such as DNA or RNA, and the nucleic acid will be combined with the magnetic beads in the solution. Then, the magnetic rod 12 is inserted to adsorb the nucleic acid combined with the magnetic beads. The lysis solution is then transferred to the waste liquid chamber 11 by the negative pressure formed in the waste liquid chamber by the first negative pressure pipeline 8. Then, a certain amount of cleaning solution is injected through the solution injection port to clean the magnetic beads.
  • the cleaning solution in the upper reaction chamber is sucked into the waste liquid chamber by negative pressure. Then, a certain volume of eluent is injected through the solution injection port, and finally the second negative pressure pipeline 10 is started to form a negative pressure in the lower reaction chamber, so that the sample in the upper reaction chamber enters the lower reaction chamber through the one-way valve 8 and is collected through the sampling hole, completing a complete set of nucleic acid collection processes.
  • an action is performed according to an element, it means that the action is performed at least according to the element, which includes two situations: performing the action only according to the element, and performing the action according to the element and other elements.
  • Expressions such as multiple, multiple, and multiple include 2, 2 times, 2 kinds, and more than 2, more than 2 times, and more than 2 kinds.

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Abstract

公开了一种全自动一体化核酸提取设备，其包括：外壳、投样孔、第一负压孔、第二负压孔、磁棒插入孔、取样孔、第一负压管道和第二负压管道；所述外壳呈方形，所述投样孔、磁棒插入孔、第一负压孔和第二负压孔被布置在所述外壳顶部，所述取样孔被布置在所述外壳侧面，所述外壳内部包括废料腔和反应腔，所述废料腔和所述反应腔通过隔板隔开，所述反应腔内部设置有单向阀。与现有技术相比，检测过程中无交叉污染，提取效率高；本身结构精简，成本相对较低，便于和核酸扩增设备整合；操作简单，不容易出错，结果重复性好且对操作者及使用的硬件设施要求不高，适用性好，可同时满足临床门诊和基层医疗机构的样品检测需求。

Description

全自动一体化核酸提取设备 技术领域
本本申请涉及核酸提取仪，特别涉及一种全自动的一体化核酸提取设备。
背景技术
准确诊断、精准治疗是临床诊疗的国际发展大趋势。无论是病因复杂的肿瘤或代谢疾病的诊疗需求，还是结构简单的病原微生物感染后的筛查，都会遵守这一准则。而临床对于诊断或筛查的追求，则以准确、快速为其永恒的目标。在确保灵敏度的前提下的，检测过程越快、检测结果越早完成，越能及时地帮助临床医生精准用药，这也是精准治疗的前提。
临床上病菌和病毒等病原微生物的检测方法主要包括镜检法、培养法、免疫学检测和核酸扩增检测等。镜检法的成本低、简单、方便，但是阳性率低，容易漏检；培养法的结果可信度高，但周期长、成本高；免疫学检测的操作相对简单，但有窗口期，故容易漏检。核酸检测灵敏度特异性与培养法相当，但也有以下的不足：①样品制备(核酸提取)、核酸扩增/检测分开进行，操作步骤繁琐，需要专业人员进行，增加了操作误差；②开放式核酸提取也容易造成样品的交叉污染，出现检测结果的假阳性；③虽然核酸检测相对于病原微生物培养操作周期较短，但也很难达到床边检测(POCT)的要求。如何使快速扩增核酸、缩短检测时间、减少病人等候时间成为目前的当务之急。
发明内容
本申请的目的在于提供一种全自动一体化核酸提取设备，用以使得设备一体化，并减少交叉污染。
本申请公开了一种全自动一体化核酸提取设备，所述提取设备包括：外壳、投样孔、第一负压孔、第二负压孔、磁棒插入孔、取样孔、第一负压管道和第二负压管道；所述外壳呈方形，所述投样孔、磁棒插入孔、第一负压孔和第二负压孔被布置在所述外壳顶部，所述取样孔被布置在所述外壳侧面，所述外壳内部包括废料腔和反应腔，所述废料腔和所述反应腔通过隔板隔开，所述反应腔内部设置有单向阀，将所述反应腔隔为上部反应腔和下部反应腔，所述上部反应腔以及所述第一负压管道与所述废料腔连接，所述第二负压管道通过所述第二负压孔与所述下部反应腔连接。
在一个优选例中，所述，投样孔包括加样孔和溶液注入孔。
在一个优选例中，所述溶液注入孔与多向泵相连，通过控制不同液体的注入以完成病原微生物的核酸提取。
在一个优选例中，所述反应腔内部设置有磁珠和磁套。
在一个优选例中，所述取样孔与所述下部反应腔连接。
在一个优选例中，所述第一负压管道和所述第二负压管道与负压泵连接。
在一个优选例中，所述设备配置有注射泵，所述注射泵与所述溶液注入孔连接。
在一个优选例中，所述设备配置有超声发生器和加热模块，所述超声发生器和加热模块紧贴所述核酸提取设备的侧面和后面。
在一个优选例中，所述磁棒插入孔被配置在所述加样孔和溶液注入孔的中间。
在一个优选例中，所述取样孔处设置有取样棒。
本发明的优点在于：
1)检测过程中无交叉污染，检测准确度高；
2)仪器本身结构精简，成本相对较低；
3)操作简单，不容易出错，结果重复性好；
4)对操作者及使用的硬件设施要求极低，适用性好，可同时满足临床门诊和基层医疗机构的样品检测需求；
5)节省时间，整个核酸提取完成操作时间不超过10分钟
本申请的说明书中记载了大量的技术特征，分布在各个技术方案中，如果要罗列出本申请所有可能的技术特征的组合(即技术方案)的话，会使得说明书过于冗长。为了避免这个问题，本申请上述发明内容中公开的各个技术特征、在下文各个实施方式和例子中公开的各技术特征、以及附图中公开的各个技术特征，都可以自由地互相组合，从而构成各种新的技术方案(这些技术方案均因视为在本说明书中已经记载)，除非这种技术特征的组合在技术上是不可行的。例如，在一个例子中公开了特征A+B+C，在另一个例子中公开了特征A+B+D+E，而特征C和D是起到相同作用的等同技术手段，技术上只要择一使用即可，不可能同时采用，特征E技术上可以与特征C相组合，则，A+B+C+D的方案因技术不可行而应当不被视为已经记载，而A+B+C+E的方案应当视为已经被记载。
附图说明
下面将对实施例或现有技术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是根据本发明的一实施例的核酸提取盒整体结构示意图；
图2是根据本发明的一实施例的核酸提取盒的侧面剖视图；
图3是根据本发明的一实施例的核酸提取盒的侧视图；
图4是根据本发明的一实施例的另一角度的核酸提取设备侧面剖视图。
附图说明：
1-加样孔，2-磁棒插入孔，3-溶液注入孔，4-第一负压孔；5-第二负压孔，6-取样孔，7-反应腔，8-单向阀，9-第一负压管道；10-第二负压管道，11-废液腔；12-磁棒。
具体实施方式
本发明的发明人经过深入研究，开发了一种全自动一体化核酸提取设备。该设备检测过程中无交叉污染，核酸提取效率高；本身结构精简，成本相对较低，便于和核酸扩增设备整合；操作简单，不容易出错，结果重复性好且对操作者及使用的硬件设施要求极低，适用性好，可同时满足临床门诊和基层医疗机构的样品检测需求；最节省时间，整个核酸提取完成操作时间不超过10分钟。
为使本申请的目的，技术方案和优点更加清楚，下面将结合附图对本申请的实施方式作进一步地详细描述。
实施例
本发明的一实施例如图1至图4所示，其包括外壳、投样孔、第一负压孔4、第二负压孔5、磁棒插入孔2、取样孔6、第一负压管道9和第二负压管道10；其中：
所述外壳呈方形，所述投样孔6、磁棒插入孔2、第一负压孔4和第二负压孔5被布置在所述外壳顶部，所述取样孔6被布置在所述外壳侧面。可选的，在一实施例中，所述投样孔包括加样孔1和溶液注入孔3。可选的，在另一实施例中，所述溶液注入孔3与多向泵相连，通过控制不同液体的注入以完成病原微生物的核酸提取。
所述外壳内部包括废料腔11和反应腔7，可选的，在一实施例中，所述反应腔7内部设置有磁珠。所述废料腔11和所述反应腔7通过隔板隔开，所述反应腔7内部设置有单向阀8，将所述反应腔7隔为上部反应腔和下部反应腔，所述上部反应腔以及所述第一负压管道9与所述废料腔11连接，所述第二负压管道10通过所述第二负压孔5与所述下部反应腔连接。可选的，在一实施例中，所述第一负压管道9和所述第二负压管道10与负压泵 连接。
可选的，在一实施例中，所述核酸提取设备还配置有配置有超声发生器和加热模块，所述超声发生器和加热模块紧贴所述核酸提取设备的侧面和后面用以在使用过程中提供超声震动和加热。
可选的，在一实施例中，所述取样孔6处设置有取样棒
可选地，在一实施例中，所述核酸提取设备还包括磁棒12，磁棒12通过所述磁棒插入孔2插入所述反应腔7,用以核酸提取,可选地，在另一实施例中，所述磁棒配置有磁套，所述磁套嵌入在所述反应腔中。
本实施例在使用过程中的流程为：先通过加样孔加入一定量的含病原微生物的痰液或漱口液等，样品与预灌装的裂解液混合，通过超声和加热的方法裂解病毒和细菌等病原微生物，释放其DNA或RNA等核酸物质，核酸将于溶液中的磁珠结合。然后，伸入磁棒12，用以吸附和磁珠结合的核酸。裂解液随后通过第一负压管道8在废液腔形成的负压将裂解液转移至废液腔11中。然后通过溶液注入口注入一定量的清洗溶液，清洗磁珠。一段时间后再通过负压使得上部反应腔中的清洗液吸入到废液腔中。然后，通过溶液注入口注入一定体积的洗脱液，最后启动第二负压管道10在下部反应腔形成负压，使得上部反应腔中的样本通过单向阀8进入下部反应腔，并通过取样孔收集，完成一整套的核酸采集流程。
需要说明的是，在本专利的申请文件中，诸如第一和第二等之类的关系术语仅仅用来将一个实体或者操作与另一个实体或操作区分开来，而不一定要求或者暗示这些实体或操作之间存在任何这种实际的关系或者顺序。而且，术语“包括”、“包含”或者其任何其他变体意在涵盖非排他性的包含，从而使得包括一系列要素的过程、方法、物品或者设备不仅包括那些要素，而且还包括没有明确列出的其他要素，或者是还包括为这种过程、方法、物品或者设备所固有的要素。在没有更多限制的情况下，由语句“包括一个”限定的要素，并不排除在包括所述要素的过程、方法、物品或者设备中还存在另外的相同要素。本专利的申请文件中，如果提到根据某要素执行某行为，则是指至少根据该要素执行该行为的意思，其中包括了两种情况：仅根据该要素执行该行为、和根据该要素和其它要素执行该行为。多个、多次、多种等表达包括2个、2次、2种以及2个以上、2次以上、2种以上。
在本申请提及的所有文献都被认为是整体性地包括在本申请的公开内容中，以便在必要时可以作为修改的依据。此外应理解，在阅读了本申请的上述公开内容之后，本领域技术人员可以对本申请作各种改动或修改，这些等价形式同样落于本申请所要求保护的范围。

Claims (10)

  1. 一种全自动一体化核酸提取设备，其特征在于，所述提取设备包括：外壳、投样孔、第一负压孔、第二负压孔、磁棒插入孔、取样孔、第一负压管道和第二负压管道；所述外壳呈方形，所述投样孔、磁棒插入孔、第一负压孔和第二负压孔被布置在所述外壳顶部，所述取样孔被布置在所述外壳侧面，所述外壳内部包括废料腔和反应腔，所述废料腔和所述反应腔通过隔板隔开，所述反应腔内部设置有单向阀，将所述反应腔隔为上部反应腔和下部反应腔，所述上部反应腔以及所述第一负压管道与所述废料腔连接，所述第二负压管道通过所述第二负压孔与所述下部反应腔连接。
  2. 如权利要求1所述的核酸提取设备，其特征在于，所述，投样孔包括加样孔和溶液注入孔。
  3. 根据权利要求2所述的核酸提取设备，其特征在于，所述溶液注入孔与多向泵相连，通过控制不同液体的注入以完成病原微生物的核酸提取。
  4. 根据权利要求1所述的核酸提取设备，其特征在于，所述反应腔内部设置有磁珠和磁套。
  5. 根据权利要求1所述的核酸提取设备，其特征在于，所述取样孔与所述下部反应腔连接。
  6. 根据权利要求1所述的核酸提取设备，其特征在于，所述第一负压管道和所述第二负压管道与负压泵连接。
  7. 根据权利要求2所述的核酸提取设备，其特征在于，所述设备配置有注射泵，所述注射泵与所述溶液注入孔连接。
  8. 根据权利要求1所述的核酸提取设备，其特征在于，所述设备配置有超声发生器和加热模块，所述超声发生器和加热模块紧贴所述核酸提取设备的侧面和后面。
  9. 根据权利要求2所述的核酸提取设备，其特征在于，所述磁棒插入孔被配置在所述加样孔和溶液注入孔的中间。
  10. 根据权利要求1所述的核酸提取设备，其特征在于，所述取样孔处设置有取样棒。
PCT/CN2024/090407 2023-04-28 2024-04-28 全自动一体化核酸提取设备 WO2024222956A1 (zh)

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