WO2023272800A1 - Carte de diagnostic moléculaire contenant un processus complet d'extraction, d'amplification et de détection, et son procédé d'utilsation - Google Patents

Carte de diagnostic moléculaire contenant un processus complet d'extraction, d'amplification et de détection, et son procédé d'utilsation Download PDF

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Publication number
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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WO
WIPO (PCT)
Prior art keywords
chamber
extraction
waste liquid
detection
delivery pipeline
Prior art date
Application number
PCT/CN2021/106981
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English (en)
Chinese (zh)
Inventor
肖博
陈铖
曾玉祥
王东伟
康建成
陆沙
闫露露
Original Assignee
北京乐普智慧医疗科技有限公司
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Application filed by 北京乐普智慧医疗科技有限公司 filed Critical 北京乐普智慧医疗科技有限公司
Publication of WO2023272800A1 publication Critical patent/WO2023272800A1/fr

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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

Definitions

  • 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 .

Abstract

Carte de diagnostic moléculaire contenant un processus complet d'extraction, d'amplification et de détection, la carte comprenant une couche supérieure de puce (1), une couche intermédiaire de puce (2) et une couche inférieure de puce (3). La couche supérieure de puce (1) est munie en son sein d'une cavité d'échantillon (4), d'une cavité d'élution (5), d'une première cavité de nettoyage (6), d'une seconde cavité de nettoyage (7), d'une cavité d'extraction (8), d'une cavité de bille magnétique (9) et d'une cavité de liquide résiduaire (10), l'extrémité supérieure de la cavité d'échantillon (4) étant munie d'un tube d'échantillonnage (11) de manière insérée ; et la cavité d'échantillon (4), la cavité d'élution (5), la deuxième cavité de nettoyage (7), la première cavité de nettoyage (6), la cavité d'extraction (8), la cavité de bille magnétique (9) et la cavité de liquide résiduaire (10) sont toutes reliées à des conduites de transport (12). La face frontale de la couche supérieure de puce (1) est dotée d'une vanne rotative (13). La couche intermédiaire de puce (2) est pourvue de trous de détection (17 à 19), les trous de détection (17 à 19) étant en liaison avec les conduites de transport (12). La carte de diagnostic moléculaire se caractérise par une structure simple et des coûts réduits, les coûts étant fortement réduits, et elle n'exige que peu d'instruments adaptés, si bien que le processus complet de diagnostic moléculaire peut être réalisé au moyen d'une simple opération. Tous les réactifs de réaction sont inclus dans la carte de diagnostic moléculaire sans avoir besoin d'être ajoutés séparément par un opérateur.
PCT/CN2021/106981 2021-07-02 2021-07-19 Carte de diagnostic moléculaire contenant un processus complet d'extraction, d'amplification et de détection, et son procédé d'utilsation WO2023272800A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202110748581.7 2021-07-02
CN202110748581 2021-07-02

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Publication Number Publication Date
WO2023272800A1 true WO2023272800A1 (fr) 2023-01-05

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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 (fr) * 2014-11-11 2016-05-19 Genmark Diagnostics, Inc. Instrument et cartouche pour effectuer des dosages dans un système de réaction et de préparation d'échantillon fermé
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 (fr) * 2014-11-11 2016-05-19 Genmark Diagnostics, Inc. Instrument et cartouche pour effectuer des dosages dans un système de réaction et de préparation d'échantillon fermé
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

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