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WO2023272800A1 - 一种包含提取扩增检测全流程的分子诊断卡及其使用工艺方法 - Google Patents

一种包含提取扩增检测全流程的分子诊断卡及其使用工艺方法 Download PDF

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WO2023272800A1
WO2023272800A1 PCT/CN2021/106981 CN2021106981W WO2023272800A1 WO 2023272800 A1 WO2023272800 A1 WO 2023272800A1 CN 2021106981 W CN2021106981 W CN 2021106981W WO 2023272800 A1 WO2023272800 A1 WO 2023272800A1
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chamber
extraction
waste liquid
detection
delivery pipeline
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肖博
陈铖
曾玉祥
王东伟
康建成
陆沙
闫露露
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Lepu Medical Technology Beijing Co Ltd
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Lepu Medical Technology Beijing Co Ltd
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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
    • C12M1/34—Measuring or testing with condition measuring or sensing means, e.g. colony counters
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions

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  • the invention relates to a molecular diagnosis card including the whole process of extraction, amplification and detection and its application process, belonging to the technical field of molecular diagnosis.
  • Molecular diagnosis refers to the technique of using molecular biological methods to detect changes in the structure or expression level of genetic material in patients to make a diagnosis.
  • Molecular diagnosis is the main method of predictive diagnosis, which can be used for diagnosis of individual genetic diseases and prenatal diagnosis.
  • Molecular diagnosis mainly refers to the detection of genes encoding various structural proteins, enzymes, antigen antibodies, and immune active molecules related to diseases.
  • This patent realizes a molecular diagnosis card including the whole process of extraction, amplification and detection, which has a simple structure and low cost, and can realize the whole process of molecular diagnosis with only simple operation.
  • the present invention provides a molecular diagnostic card including the whole process of extraction, amplification and detection and its application process.
  • the molecular diagnostic card has a simple structure, low cost, and greatly reduces the cost. The requirements are low, and the whole process of molecular diagnosis can be realized with simple operation.
  • a molecular diagnostic card including the whole process of extraction, amplification and detection, the molecular diagnostic card includes the upper layer of the chip, the middle layer of the chip and the lower layer of the chip, and the rear end surface of the upper layer of the chip is fixedly connected The middle layer of the chip, the rear end surface of the middle layer of the chip is fixedly connected to the lower layer of the chip;
  • the upper layer of the chip is provided with a sample chamber, an elution chamber, a cleaning chamber, a cleaning chamber, an extraction chamber, a magnetic bead chamber and a waste liquid chamber.
  • the upper end of the sample chamber is plugged with a sampling tube.
  • the sample Chamber, elution chamber, cleaning chamber 2, cleaning chamber 1, extraction chamber, magnetic bead chamber and waste liquid chamber are all connected with delivery pipelines, the front end of the upper layer of the chip is equipped with a rotary valve, and the middle layer of the chip is equipped with a detection valve. hole, and the detection hole is connected with a delivery pipeline.
  • the sample chamber is used to add samples
  • the eluent is stored in the elution chamber
  • the cleaning solution 2 is stored in the cleaning chamber 2
  • the cleaning liquid 1 is stored in the cleaning chamber 1
  • the extraction chamber is used as the extraction operation area
  • the magnetic bead liquid is stored in the magnetic bead chamber
  • the waste liquid chamber is used for waste liquid discharge
  • each delivery pipeline is controlled by a rotary valve to realize the on-off between each chamber.
  • the front end of the upper layer of the chip is clamped with a reaction reagent sealing layer, and the upper end of the upper layer of the chip is clamped with an extraction reagent sealing layer.
  • a detection sealing layer is clamped at the detection hole of the middle layer of the chip.
  • the detection hole includes a first detection hole, a second detection hole and a third detection hole, and the arrangement of multiple detection holes can realize the simultaneous detection of different parameters of the same sample and improve work efficiency.
  • a sealing gasket is provided between the rotary valve and the upper layer of the chip, and a plurality of through holes are arranged on the sealing gasket, and the through holes are communicated with the delivery pipeline, and rotating the rotary valve can control the flow of each channel on the sealing gasket.
  • the rotary valve includes an air valve and a liquid valve
  • the liquid valve can control the delivery of liquid between the delivery pipelines
  • the air valve controls the on-off of the gas to provide driving force for the delivery of the liquid.
  • the middle layer of the chip is provided with three air holes, and the three air holes communicate with the extraction chamber, the rotary valve and the waste liquid chamber respectively, and a filter membrane is attached on the surface of the air holes.
  • the air hole communicates with the extraction chamber to provide gas pressure for the extraction chamber to facilitate liquid transportation; the air hole communicates with the air valve of the rotary valve to provide gas pressure for the air valve; the air hole communicates with the waste liquid chamber to facilitate the discharge of waste gas; in addition
  • a layer of filter membrane is attached to the surface of the vent hole, and different filter membrane pore sizes can be used to filter aerosols or pathogens of different sizes.
  • the delivery pipeline connected to the detection hole is provided with a quantitative sensor
  • the delivery pipeline connected to the detection hole is provided with a reaction reagent hole
  • the reaction reagent hole is located between the detection hole and the quantitative sensor on the delivery pipeline.
  • the nucleic acid is quantified by the quantitative sensor before entering the detection hole, which is more conducive to the accuracy and convenience of relevant data detection; the reaction reagent is stored in the reaction reagent hole, which can directly react with the quantified nucleic acid, which is more convenient and does not need to manually add the reaction reagent .
  • the present invention also discloses a process method for using the molecular diagnostic card including the whole process of extraction, amplification and detection.
  • the process method includes the following steps:
  • the molecular diagnosis card has a simple structure, low cost, greatly reduces the cost, has low requirements for supporting instruments, and can realize the whole process of molecular diagnosis only by simple operation. All the reagents used in the reaction are included in the molecular diagnostic card, and no operator needs to add them separately.
  • Fig. 1 is the structural representation of the molecular diagnostic card described in the embodiment
  • Fig. 2 is the front view of the molecular diagnostic card described in the embodiment
  • Fig. 3 is the rear view of the molecular diagnostic card described in the embodiment.
  • Fig. 4 is the front view of the upper layer of the chip described in the embodiment.
  • Fig. 5 is the front view of the middle layer of the chip described in the embodiment.
  • Fig. 6 is the structural schematic diagram of gasket described in the embodiment.
  • Fig. 7 is a flow chart of the use process of the molecular diagnostic card described in the embodiment.
  • a molecular diagnostic card including the whole process of extraction, amplification and detection is characterized in that the molecular diagnostic card includes a chip upper layer 1, a chip middle layer 2 and a chip lower layer 3, and the rear end surface of the chip upper layer 1 is fixedly connected to the middle layer 2 of the chip, and the rear end surface of the middle layer 2 of the chip is fixedly connected to the lower layer 3 of the chip;
  • the upper layer 1 of the chip is provided with a sample chamber 4, an elution chamber 5, a cleaning chamber 6, a cleaning chamber 7, an extraction chamber 8, a magnetic bead chamber 9 and a waste liquid chamber 10.
  • the upper end of the sample chamber 4 is inserted A sampling tube 11 is connected, and the sample chamber 4, the elution chamber 5, the second cleaning chamber 7, the first cleaning chamber 6, the extraction chamber 8, the magnetic bead chamber 9 and the waste liquid chamber 10 are all connected with a delivery pipeline 12 (Fig. 4, the delivery pipeline 12 connected to the extraction cavity 8 is located at the rear side of the extraction cavity 8, and Fig.
  • FIG. 4 is a front view of the upper layer 1 of the chip, so the delivery pipeline 12 connected to the extraction cavity 8 cannot be seen), the upper layer of the chip A rotary valve 13 is installed on the front end of the chip 1, and a detection hole is provided in the middle layer 2 of the chip, and a delivery pipeline 12 is connected to the detection hole.
  • the front end of the upper chip layer 1 is clamped with a reaction reagent sealing layer 14 , and the upper end of the chip upper layer 1 is clamped with an extraction reagent sealing layer 15 .
  • a detection sealing layer 16 is clamped at the detection hole of the middle layer 2 of the chip.
  • the detection holes include a first detection hole 17 , a second detection hole 18 and a third detection hole 19 .
  • a gasket 21 is provided between the rotary valve 13 and the chip upper layer 1 , and a plurality of through holes 22 are provided on the gasket 21 , and the through holes 22 communicate with the delivery pipeline 12 .
  • the rotary valve 13 includes a gas valve 13-1 and a liquid valve 13-2.
  • the middle layer 2 of the chip is provided with three air holes 20, and the three air holes 20 communicate with the extraction chamber 8, the rotary valve 13 and the waste liquid chamber 10 respectively, and a filter membrane is attached on the surface of the air holes 20.
  • a quantitative sensor 23 is provided on the delivery pipeline 12 connected to the detection hole, and a reaction reagent hole 24 is provided on the delivery pipeline 12 connected to the detection hole, and the reaction reagent hole 24 is located between the detection hole and the detection hole. On the delivery pipeline 12 between the quantitative sensors 23 .

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Abstract

一种包含提取扩增检测全流程的分子诊断卡，包括芯片上层(1)、芯片中层(2)和芯片下层(3)，芯片上层(1)内部设有样本腔(4)、洗脱腔(5)、清洗一腔(6)、清洗二腔(7)、提取腔(8)、磁珠腔(9)和废液腔(10)，样本腔(4)上端插接有采样管(11)，样本腔(4)、洗脱腔(5)、清洗二腔(7)、清洗一腔(6)、提取腔(8)、磁珠腔(9)和废液腔(10)均连接有输送管路(12)，芯片上层(1)的前端面安装有旋转阀(13)；芯片中层(2)设有检测孔(17-19)，检测孔(17-19)连接有输送管路(12)。分子诊断卡结构简单，成本低，极大降低成本，对配套仪器要求低，仅需简易的操作便可实现全流程的分子诊断。所有反应用试剂均包含在分子诊断卡内，无需操作人员单独添加。

Description

一种包含提取扩增检测全流程的分子诊断卡及其使用工艺方法 技术领域
本发明涉及一种包含提取扩增检测全流程的分子诊断卡及其使用工艺方法，属于分子诊断技术领域。
背景技术
分子诊断是指应用分子生物学方法检测患者体内遗传物质的结构或表达水平的变化而做出诊断的技术。分子诊断是预测诊断的主要方法，既可以进行个体遗传病的诊断，也可以进行产前诊断。分子诊断主要是指编码与疾病相关的各种结构蛋白、酶、抗原抗体、免疫活性分子基因的检测。
现有的分子诊断需要大量的人工操作步骤，市场上急需便于操作使用的分子诊断产品，但是市面上已有的产品往往产品复杂导致成本高昂，或者依然需要人工操作，无法实现自动化。原材料成本高，生产难度大，配套仪器复杂。例如专利US6783934中公开的技术，需要通过机械运行活塞驱动进行核酸提取，配套仪器需要机械运动，检测试剂需要外加，无法实现自动化。
本专利实现了一种包含提取扩增检测全流程的分子诊断卡，结构简单，成本低，仅需简易的操作便可实现全流程的分子诊断。
发明内容
本发明针对现有技术存在的不足，提供一种包含提取扩增检测全流程的分子诊断卡及其使用工艺方法，所述的分子诊断卡结构简单，成本低，极大降低成本，对配套仪器要求低，仅需简易的操作便可实现全流程的分子诊断。
本发明解决上述技术问题的技术方案如下：一种包含提取扩增检测全流程的分子诊断卡，所述的分子诊断卡包括芯片上层、芯片中层和芯片下层，所述的芯片上层后端面固定连接所述芯片中层，所述芯片中层的后端面固定连接所述芯片下层；
所述的芯片上层内部设有样本腔、洗脱腔、清洗一腔、清洗二腔、提取腔、磁珠腔和废液腔，所述的样本腔上端插接有采样管，所述的样本腔、洗脱腔、清洗二腔、清洗一腔、提取腔、磁珠腔和废液腔均连接有输送管路，所述芯片上层的前端面安装有旋转阀，所述芯片中层设有检测孔，所述检测孔连接有输送管路。样本腔内用于加入样本，洗脱腔内存放洗脱液，清洗二腔内存放清洗液二，清洗一腔内存放清洗液一，提取腔作为提取运行区域，磁珠腔内存放磁珠液，废液腔用于废液排入，通过旋转阀控制各个输送管路实现各个腔体之间的通断。
进一步的，所述的芯片上层的前端面卡接有反应试剂密封层，所述芯片上层的上端卡接有提取试剂密封层。
进一步的，所述芯片中层的检测孔处卡接有检测密封层。
进一步的，所述的检测孔包括第一检测孔、第二检测孔和第三检测孔，多个检测孔的设置可以实现同一样本不同参数的同时检测，提高工作效率。
进一步的，所述旋转阀与所述芯片上层之间设有密封垫，密封垫上设有若干通孔，所述通孔与所述输送管路相通，旋转所述旋转阀可以控制密封垫上各个通孔与相应的输送管路的通断。
进一步的，所述旋转阀包括气阀和液阀，液阀可以控制输送管路之间液体的输送，气阀控器气体的通断，为液体输送提供推动力。
进一步的，所述的芯片中层设有三个透气孔，三个所述透气孔分别与所述提取腔、旋转阀和废液腔相通，所述透气孔表面附有过滤膜。透气孔与提取腔相通，为提取腔提供气体压力，便于液体的输送；透气孔与旋转阀的气阀相通，为气阀提供气体压力；透气孔与废液腔相通，便于废气的排出；另外透气孔表面附加一层过滤膜，使用不同的过滤膜孔径可以用于过滤不同大小的气溶胶或者病原体。
进一步的，与所述检测孔连接的输送管路上设有定量传感器，所述检测孔连接的输送管路上设有反应试剂孔，所述反应试剂孔位于所述检测孔和所述定量传感器之间的输送管路上。核酸进入检测孔前经过定量传感器进行定量，更利于相关数据检测的准确性和便捷性；反应试剂孔内存放有反应试剂，可以直接与定量后的核酸进行反应，更加便捷，无需人工加入反应试剂。
本发明还公开了包含提取扩增检测全流程的分子诊断卡的使用工艺方法，所述的工艺方法包括如下步骤：
(1)调节旋转阀，所述提取腔与所述磁珠腔通过输送管路导通，磁珠进入提取腔；
(2)调节旋转阀，所述提取腔与所述废液腔通过输送管路导通，提取腔内的废液排入废液腔；
(3)调节旋转阀，所述提取腔与所述样本腔通过输送管路导通，样本进入提取腔，磁珠吸附样本核酸；
(4)调节旋转阀，所述提取腔与所述废液腔通过输送管路导通，提取腔内的废液排入废液腔；
(5)调节旋转阀，所述提取腔与所述清洗一腔通过输送管路导通，所述清洗一腔 内的清洗液进入提取腔，清洗磁珠吸附的非核酸物质；
(6)调节旋转阀，所述提取腔与所述废液腔通过输送管路导通，提取腔内的废液排入废液腔；
(7)调节旋转阀，所述提取腔与所述清洗二腔通过输送管路导通，所述清洗二腔内的清洗液进入提取腔，再次清洗磁珠吸附的非核酸物质；
(8)调节旋转阀，所述提取腔与所述废液腔通过输送管路导通，提取腔内的废液排入废液腔；
(9)调节旋转阀，所述提取腔与所述洗脱腔通过输送管路导通，洗脱液进入所述提取腔洗脱磁珠吸附的目的核酸物质；
(10)调节旋转阀，所述提取腔与所述检测孔通过输送管路导通，洗脱的核酸进入检测孔前进行定量检测；
(11)调节旋转阀，所述提取腔与所述废液腔通过输送管路导通，剩余的核酸排入废液腔；
(12)调节旋转阀，所述提取腔与所述检测孔通过输送管路导通，定量后核酸进入检测孔，同时加入反应试剂，与仪器配合进行核酸扩增、检测。
本发明的有益效果是：所述的分子诊断卡结构简单，成本低，极大降低成本，对配套仪器要求低，仅需简易的操作便可实现全流程的分子诊断。所有反应用的试剂均包含在分子诊断卡内，无需操作人员单独添加。
附图说明
图1为实施例中所述分子诊断卡的结构示意图；
图2为实施例中所述分子诊断卡的主视图；
图3为实施例中所述分子诊断卡的后视图；
图4为实施例中所述芯片上层的主视图；
图5为实施例中所述芯片中层的主视图；
图6为实施例中所述密封垫的结构示意图；
图7为实施例中所述分子诊断卡的使用工艺流程图；
图中，1芯片上层，2芯片中层，3芯片下层，4样本腔，5洗脱腔，6清洗一腔，7清洗二腔，8提取腔，9磁珠腔，10废液腔，11采样管，12输送管路，13旋转阀，13-1气阀，13-2液阀，14反应试剂密封层，15提取试剂密封层，16检测密封层，17第一检测孔，18第二检测孔，19第三检测孔，20透气孔，21密封垫，22通孔，23定量传 感器，24反应试剂孔。
具体实施方式
为使本发明的上述目的、特征和优点能够更加明显易懂，下面对本发明的具体实施方式做详细的说明。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。但是本发明能够以很多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改进，因此本发明不受下面公开的具体实施例的限制。
除非另有定义，本文所使用的所有的技术和科学术语与属于本发明的技术领域的技术人员通常理解的含义相同。本文中在本发明的说明书中所使用的术语只是为了描述具体的实施方式的目的，不是旨在于限制本发明。
如图所示，一种包含提取扩增检测全流程的分子诊断卡，其特征在于，所述的分子诊断卡包括芯片上层1、芯片中层2和芯片下层3，所述的芯片上层1后端面固定连接所述芯片中层2，所述芯片中层2的后端面固定连接所述芯片下层3；
所述的芯片上层1内部设有样本腔4、洗脱腔5、清洗一腔6、清洗二腔7、提取腔8、磁珠腔9和废液腔10，所述的样本腔4上端插接有采样管11，所述的样本腔4、洗脱腔5、清洗二腔7、清洗一腔6、提取腔8、磁珠腔9和废液腔10均连接有输送管路12(图4中提取腔8连接的输送管路12位于所述提取腔8的后侧面，图4是芯片上层1的主视图，所以看不到提取腔8连接的输送管路12)，所述芯片上层1的前端面安装有旋转阀13，所述芯片中层2设有检测孔，所述检测孔连接有输送管路12。
所述的芯片上层1的前端面卡接有反应试剂密封层14，所述芯片上层1的上端卡接有提取试剂密封层15。所述芯片中层2的检测孔处卡接有检测密封层16。所述的检测孔包括第一检测孔17、第二检测孔18和第三检测孔19。所述旋转阀13与所述芯片上层1之间设有密封垫21，密封垫21上设有若干通孔22，所述通孔22与所述输送管路12相通。所述旋转阀13包括气阀13-1和液阀13-2。所述的芯片中层2设有三个透气孔20，三个所述透气孔20分别与所述提取腔8、旋转阀13和废液腔10相通，所述透气孔20表面附有过滤膜。与所述检测孔连接的输送管路12上设有定量传感器23，所述检测孔连接的输送管路12上设有反应试剂孔24，所述反应试剂孔24位于所述检测孔和所述定量传感器23之间的输送管路12上。
所述包含提取扩增检测全流程的分子诊断卡的使用工艺方法，包括如下步骤：
(1)调节旋转阀13，所述提取腔8与所述磁珠腔9通过输送管路12导通，磁珠进入提取腔8；
(2)调节旋转阀13，所述提取腔8与所述废液腔10通过输送管路12导通，提取腔8内的废液排入废液腔10；
(3)调节旋转阀13，所述提取腔8与所述样本腔4通过输送管路12导通，样本进入提取腔8，磁珠吸附样本核酸；
(4)调节旋转阀13，所述提取腔8与所述废液腔10通过输送管路12导通，提取腔8内的废液排入废液腔10；
(5)调节旋转阀13，所述提取腔8与所述清洗一腔6通过输送管路12导通，所述清洗一腔6内的清洗液进入提取腔8，清洗磁珠吸附的非核酸物质；
(6)调节旋转阀13，所述提取腔8与所述废液腔10通过输送管路12导通，提取腔8内的废液排入废液腔10；
(7)调节旋转阀13，所述提取腔8与所述清洗二腔7通过输送管路12导通，所述清洗二腔7内的清洗液进入提取腔8，再次清洗磁珠吸附的非核酸物质；
(8)调节旋转阀13，所述提取腔8与所述废液腔10通过输送管路12导通，提取腔8内的废液排入废液腔10；
(9)调节旋转阀13，所述提取腔8与所述洗脱腔5通过输送管路12导通，洗脱液进入所述提取腔8洗脱磁珠吸附的目的核酸物质；
(10)调节旋转阀13，所述提取腔8与所述第三检测孔19通过输送管路12导通，洗脱的核酸进入第三检测孔19前进行定量检测；
(11)调节旋转阀13，所述提取腔8与所述第二检测孔18通过输送管路12导通，洗脱的核酸进入第二检测孔18前进行定量检测；
(12)调节旋转阀13，所述提取腔8与所述第一检测孔17通过输送管路12导通，洗脱的核酸进入第一检测孔17前进行定量检测；
(13)调节旋转阀13，所述提取腔8与所述废液腔10通过输送管路12导通，剩余的核酸排入废液腔10；
(14)调节旋转阀13，所述提取腔8与所述检测孔通过输送管路12导通，定量后核酸进入第一检测孔17检测孔，同时加入反应试剂，与仪器配合进行核酸扩增、检测；
(15)调节旋转阀13，所述提取腔8与所述检测孔通过输送管路12导通，定量后核酸进入第二检测孔18检测孔，同时加入反应试剂，与仪器配合进行核酸扩增、检测；
(16)调节旋转阀13，所述提取腔8与所述检测孔通过输送管路12导通，定量后核酸进入第三检测孔19检测孔，同时加入反应试剂，与仪器配合进行核酸扩增、检测。
以上所述实施例的各技术特征可以进行任意的组合，为使描述简洁，未对上述实施例中的各个技术特征所有可能的组合都进行描述，然而，只要这些技术特征的组合不存在矛盾，都应当认为是本说明书记载的范围。
以上所述实施例仅表达了本发明的几种实施方式，其描述较为具体和详细，但并不能因此而理解为对发明专利范围的限制。应当指出的是，对于本领域的普通技术人员来说，在不脱离本发明构思的前提下，还可以做出若干变形和改进，这些都属于本发明的保护范围。因此，本发明专利的保护范围应以所附权利要求为准。

Claims (9)

  1. 一种包含提取扩增检测全流程的分子诊断卡，其特征在于，所述的分子诊断卡包括芯片上层(1)、芯片中层(2)和芯片下层(3)，所述的芯片上层(1)后端面固定连接所述芯片中层(2)，所述芯片中层(2)的后端面固定连接所述芯片下层(3)；
    所述的芯片上层(1)内部设有样本腔(4)、洗脱腔(5)、清洗一腔(6)、清洗二腔(7)、提取腔(8)、磁珠腔(9)和废液腔(10)，所述的样本腔(4)上端插接有采样管(11)，所述的样本腔(4)、洗脱腔(5)、清洗二腔(7)、清洗一腔(6)、提取腔(8)、磁珠腔(9)和废液腔(10)均连接有输送管路(12)，所述芯片上层(1)的前端面安装有旋转阀(13)；所述芯片中层(2)设有检测孔，所述检测孔连接有输送管路(12)。
  2. 根据权利要求1所述的一种包含提取扩增检测全流程的分子诊断卡，其特征在于，所述的芯片上层(1)的前端面卡接有反应试剂密封层(14)，所述芯片上层(1)的上端卡接有提取试剂密封层(15)。
  3. 根据权利要求1所述的一种包含提取扩增检测全流程的分子诊断卡，其特征在于，所述芯片中层(2)的检测孔处卡接有检测密封层(16)。
  4. 根据权利要求3所述的一种包含提取扩增检测全流程的分子诊断卡，其特征在于，所述的检测孔包括第一检测孔(17)、第二检测孔(18)和第三检测孔(19)。
  5. 根据权利要求1所述的一种包含提取扩增检测全流程的分子诊断卡，其特征在于，所述旋转阀(13)与所述芯片上层(1)之间设有密封垫(21)，密封垫(21)上设有若干通孔(22)，所述通孔(22)与所述输送管路(12)相通。
  6. 根据权利要求5所述的一种包含提取扩增检测全流程的分子诊断卡，其特征在于，所述旋转阀(13)包括气阀(13-1)和液阀(13-2)。
  7. 根据权利要求1所述的一种包含提取扩增检测全流程的分子诊断卡，其特征在于，所述的芯片中层(2)设有三个透气孔(20)，三个所述透气孔(20)分别与所述提取腔(8)、旋转阀(13)和废液腔(10)相通，所述透气孔(20)表面附有过滤膜。
  8. 根据权利要求1所述的一种包含提取扩增检测全流程的分子诊断卡，其特征在于，与所述检测孔连接的输送管路(12)上设有定量传感器(23)，所述检测孔连接的输送管路(12)上设有反应试剂孔(24)，所述反应试剂孔(24)位于所述检测孔和所述定量传感器(23)之间的输送管路(12)上。
  9. 一种根据权利要求1-8任意一项所述的一种包含提取扩增检测全流程的分子诊断卡的使用工艺方法，其特征在于，所述的工艺方法包括如下步骤：
    1)调节旋转阀(13)，所述提取腔(8)与所述磁珠腔(9)通过输送管路(12)导通，磁珠进入提取腔(8)；
    2)调节旋转阀(13)，所述提取腔(8)与所述废液腔(10)通过输送管路(12)导通，提取腔(8)内的废液排入废液腔(10)；
    3)调节旋转阀(13)，所述提取腔(8)与所述样本腔(4)通过输送管路(12)导通，样本进入提取腔(8)，磁珠吸附样本核酸；
    4)调节旋转阀(13)，所述提取腔(8)与所述废液腔(10)通过输送管路(12)导通，提取腔(8)内的废液排入废液腔(10)；
    5)调节旋转阀(13)，所述提取腔(8)与所述清洗一腔(6)通过输送管路(12)导通，所述清洗一腔(6)内的清洗液进入提取腔(8)，清洗磁珠吸附的非核酸物质；
    6)调节旋转阀(13)，所述提取腔(8)与所述废液腔(10)通过输送管路(12)导通，提取腔(8)内的废液排入废液腔(10)；
    7)调节旋转阀(13)，所述提取腔(8)与所述清洗二腔(7)通过输送管路(12)导通，所述清洗二腔(7)内的清洗液进入提取腔(8)，再次清洗磁珠吸附的非核酸物质；
    8)调节旋转阀(13)，所述提取腔(8)与所述废液腔(10)通过输送管路(12)导通，提取腔(8)内的废液排入废液腔(10)；
    9)调节旋转阀(13)，所述提取腔(8)与所述洗脱腔(5)通过输送管路(12)导通，洗脱液进入所述提取腔(8)洗脱磁珠吸附的目的核酸物质；
    10)调节旋转阀(13)，所述提取腔(8)与所述检测孔通过输送管路(12)导通，洗脱的核酸进入检测孔前进行定量检测；
    11)调节旋转阀(13)，所述提取腔(8)与所述废液腔(10)通过输送管路(12)导通，剩余的核酸排入废液腔(10)；
    12)调节旋转阀(13)，所述提取腔(8)与所述检测孔通过输送管路(12)导通，定量后核酸进入检测孔，同时加入反应试剂，与仪器配合进行核酸扩增、检测。
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN118546760A (zh) * 2023-02-27 2024-08-27 上海微创惟微诊断技术有限公司 一种核酸检测试剂盒

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102459565A (zh) * 2009-06-02 2012-05-16 尹特根埃克斯有限公司 具有隔膜阀的流控设备
CN105349401A (zh) * 2015-10-14 2016-02-24 安徽易康达光电科技有限公司 一种多功能的集成化微流控核酸分析芯片及制备和分析方法
WO2016077364A2 (en) * 2014-11-11 2016-05-19 Genmark Diagnostics, Inc. Instrument and cartridge for performing assays in a closed sample preparation and reaction system
US9599610B2 (en) * 2012-12-19 2017-03-21 Dnae Group Holdings Limited Target capture system
CN110452802A (zh) * 2019-08-07 2019-11-15 深圳市刚竹医疗科技有限公司 全提取分子诊断微流控芯片及微流控系统
CN110964715A (zh) * 2019-12-05 2020-04-07 广州万孚生物技术股份有限公司 体外诊断分析装置及试剂卡
CN112063512A (zh) * 2020-09-01 2020-12-11 北京乐普智慧医疗科技有限公司 一种包含提取扩增检测全流程的分子诊断卡及其使用工艺方法

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102459565A (zh) * 2009-06-02 2012-05-16 尹特根埃克斯有限公司 具有隔膜阀的流控设备
US9599610B2 (en) * 2012-12-19 2017-03-21 Dnae Group Holdings Limited Target capture system
WO2016077364A2 (en) * 2014-11-11 2016-05-19 Genmark Diagnostics, Inc. Instrument and cartridge for performing assays in a closed sample preparation and reaction system
CN105349401A (zh) * 2015-10-14 2016-02-24 安徽易康达光电科技有限公司 一种多功能的集成化微流控核酸分析芯片及制备和分析方法
CN110452802A (zh) * 2019-08-07 2019-11-15 深圳市刚竹医疗科技有限公司 全提取分子诊断微流控芯片及微流控系统
CN110964715A (zh) * 2019-12-05 2020-04-07 广州万孚生物技术股份有限公司 体外诊断分析装置及试剂卡
CN112063512A (zh) * 2020-09-01 2020-12-11 北京乐普智慧医疗科技有限公司 一种包含提取扩增检测全流程的分子诊断卡及其使用工艺方法

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN118546760A (zh) * 2023-02-27 2024-08-27 上海微创惟微诊断技术有限公司 一种核酸检测试剂盒

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