Kit for nucleic acid analysis under totally-enclosed condition, and device and analysis method for nucleic acid analysis

A nucleic acid analysis, fully enclosed technology, applied in the field of microfluidic analysis, can solve the problems of high process requirements, complex structure of disposable reagent kits, limited promotion and application, etc.

Active Publication Date: 2020-08-07
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these products still have the problems of complex structure of disposable reagent kits, high process requirements, and high prices. The

Method used

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  • Kit for nucleic acid analysis under totally-enclosed condition, and device and analysis method for nucleic acid analysis
  • Kit for nucleic acid analysis under totally-enclosed condition, and device and analysis method for nucleic acid analysis
  • Kit for nucleic acid analysis under totally-enclosed condition, and device and analysis method for nucleic acid analysis

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Experimental program
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Effect test

Embodiment 1

[0102] Such as figure 1 Shown is a device designed for nucleic acid analysis under fully enclosed conditions in a preferred embodiment of the present invention. The main components such as the box body 1, the flexible upper cover 2, the sealing cover of the sample inlet 3 and the nucleic acid extraction probe 4 constitute the entire sealed system. The sample introduction-extraction container 6 preloaded with reagents, the nucleic acid cleaning container 7 and the nucleic acid elution-amplification container 8 are directly arranged on the box body 1, and a sealed cavity is formed between the box body 1 and the flexible upper cover 2, and the nucleic acid The extraction probe, the sample introduction-extraction container preloaded with reagents, the nucleic acid cleaning container and the nucleic acid elution-amplification container, the box body and the flexible upper cover together constitute a single-use kit. The box body 1 and the nucleic acid extraction probe 4 are made of...

Embodiment 2

[0117] Figure 2bIt is a structural schematic diagram of a typical structure of the nucleic acid extraction probe 4. The probe includes four structural and functional modules, namely the skeleton part, the mobile station joint part 41, the closed cavity joint part 42, the container sealing part 43, and the nucleic acid joint part 44. And the reagent sealing film breaking part 45. A membrane material capable of binding to DNA is bound to the nucleic acid binding portion 44 . Reagent sealing film breaking part 45 is formed by the tapered structure at the lower end of the probe skeleton itself, and is used to puncture the sealing films on the sample introduction-extraction container 6, nucleic acid cleaning container 7, and nucleic acid elution-amplification container 8. The mobile platform coupling part 41 is used to realize the combination of the probe skeleton and the mobile platform, realize the mutual fixation between the probe and the mobile platform, and the mobile platfo...

Embodiment 3

[0120] Such as Figure 5 As shown, Embodiment 3 is a further simplification in structure of Embodiment 1. The main improvement is that the nucleic acid extraction probe 4 has added an integrated sealing cover 43a of a container with a closed function. The sealing cover 43a includes 3 subunits, respectively It is used to seal the sample introduction-extraction container 6, nucleic acid cleaning container 7, and acid elution-amplification container 8 during the storage and transportation of the kit and the amplification reaction process, so no additional sealing film is needed to seal the container in this embodiment. The catchy mouth is closed. The usage method of this embodiment is basically the same as that of Embodiment 1, and the probe is placed in the probe storage container 15 before testing. Only in the nucleic acid analysis process, the step of tearing down the sealing membrane of the container is omitted, and the sealing membrane 80 in the nucleic acid elution-amplifi...

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Abstract

The invention discloses a kit capable of carrying out nucleic acid analysis under a totally-enclosed condition. The kit comprises a box body provided with a sample inlet and a sealing cover, a flexible upper cover in sealed butt joint with the box body to form a sealed cavity, one or more probes, and one or more functional containers. The sample inlet is positioned outside the sealed cavity; the sampling ends of the one or more probes are arranged in the sealed cavity, and the peripheral sides of the probes are connected with the flexible upper cover in a sealed mode; and at least one functional container is communicated with the sample inlet; and corresponding functional reagents are contained in one or more containers. The invention also provides a device and a method for nucleic acid analysis by using the kit. The nucleic acid extraction probe, the analysis reagent, the corresponding functional container and the like are sealed in independent space, so that a problem of cross contamination in the processes of sample pretreatment, transfer, amplification reaction and the like can be effectively solved, and the kit is suitable for multiple application fields of disease diagnosis,water quality detection, air monitoring, gene analysis, microbial research, pathogen detection and the like.

Description

technical field [0001] The invention relates to the field of microfluidic analysis, in particular to a device and an analysis method for nucleic acid analysis using microfluidic technology under fully enclosed conditions. Background technique [0002] Nucleic acid analysis technology, as a powerful molecular diagnostic technology, has important applications in pathogen diagnosis, infectious disease prevention, disease monitoring and treatment, etc. Compared with conventional immunological diagnostic methods, nucleic acid analysis has outstanding advantages such as high specificity and high sensitivity. Due to the low concentration of most target genes, it is difficult to directly analyze and detect, so nucleic acid analysis methods such as PCR and LAMP based on the principle of nucleic acid amplification have become the main means of nucleic acid analysis. The complete process usually includes three steps: nucleic acid extraction, nucleic acid amplification, and nucleic aci...

Claims

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Application Information

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IPC IPC(8): C12M1/00C12M1/34C12M1/36
CPCC12N15/10C12Q1/68
Inventor 方群潘建章石倩暖王慧峰王宇
Owner ZHEJIANG UNIV
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