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Anti-pollution consumable and method for carrying out CRISPR molecular diagnosis by applying anti-pollution consumable

An anti-pollution and consumable technology, applied in the field of anti-pollution consumables and CRISPR molecular diagnosis, can solve problems such as laboratory aerosol pollution, and achieve the effect of improving efficiency and simplifying operation steps

Inactive Publication Date: 2021-06-01
XI AN JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In view of this, the object of the present invention is to solve the problem of laboratory aerosol pollution caused by operations such as opening the cover, transferring sample nucleic acid, and closing the cover during the nucleic acid detection process based on CRISPR technology; the anti-pollution provided by the present invention The consumables can ensure that the nucleic acid detection based on CRISPR technology does not reduce the sensitivity and cause high false positives, so that the detection system has high robustness, reduces experimental operations, prevents pollution, and improves the efficiency of nucleic acid detection

Method used

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  • Anti-pollution consumable and method for carrying out CRISPR molecular diagnosis by applying anti-pollution consumable

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Embodiment 1

[0062] The specific structure of each part of the anti-pollution consumables provided in this embodiment includes size and preparation method.

[0063] 1. The material of the PCR outer tube is made of high-quality polypropylene material. Put the high-quality polypropylene material into the injection molding machine, heat and melt it through the injection molding machine, squeeze it into the cavity of the mold with screw pressure, and process it through cooling and molding. Suitable for conventional PCR reactions, its specific size is as follows figure 1 As shown, the unit is mm; the entity diagram is shown in figure 2 shown.

[0064] 2. The material of the reaction inner tube is made of high-quality PMMA material. Put the high-quality PMMA material into the injection molding machine, heat and melt it through the injection molding machine, squeeze it into the cavity of the mold with screw pressure, and process it through cooling and molding. There is a drain hole on one side o...

Embodiment 2

[0068] The structure of the anti-pollution consumables provided in this embodiment and the schematic diagram of each step are as follows Figure 9 ~ Figure 14 As shown, the operation steps are as follows: first, add the prepared CRISPR reagent 6 into the outer reaction tube 2, and then add the sleeve 4 into the outer reaction tube 2. The tube sleeve 4 will be stuck in the inside of the reaction outer tube 2 and fixed. Add the configured RPA nucleic acid amplification reagent 7 without sample nucleic acid into the reaction inner tube 3. During this process, try to add the RPA nucleic acid amplification reagent 7 to the bottom of the reaction inner tube 3. Due to the droplet surface tension and atmospheric pressure Strong effect, although there is a drainage hole at the bottom of the reaction inner tube 3, the RPA nucleic acid amplification reagent 7 will not flow out along the drainage hole 5. Subsequently, the reaction inner tube 3 is put into the reaction outer tube 2 to rea...

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Abstract

The invention provides an anti-pollution consumable and a method for carrying out CRISPR molecular diagnosis nucleic acid detection by applying the anti-pollution consumable, and belongs to the technical field of nucleic acid detection and molecular diagnose. The anti-pollution consumable comprises a reaction outer tube, a sleeve and a reaction inner tube; the reaction inner tube sequentially comprises a second hollow cylindrical upper body and a second conical lower body from top to bottom; the top end of the second hollow cylindrical upper body extends outwards to be connected with a fixed circular ring perpendicular to the second hollow cylindrical upper body; a plurality of liquid drainage holes are formed in the bottom of the second-type conical lower body; the diameter of the liquid discharge hole is 0.01 to 0.8 mm; the drainage holes are subjected to hydrophobic treatment; and the reaction inner tube is fixed inside the reaction outer tube through a sleeve. According to the anti-pollution consumable provided by the invention, on the premise of ensuring that nucleic acid detection based on a CRISPR technology does not reduce sensitivity and does not cause relatively high false positive, experimental operations are reduced, pollution is prevented, and the efficiency and accuracy of nucleic acid detection are improved.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid molecular diagnosis, and in particular relates to an anti-pollution consumable and a method for CRISPR molecular diagnosis using the anti-pollution consumable. Background technique [0002] Rapid, accurate, sensitive, and quantitative detection of specific nucleic acid sequences is playing an increasingly important role in the diagnosis of human infectious diseases, food security, determination of pathogens, global biosecurity, and tracking of biological pollution in environmental analysis and monitoring of environmental quality. . The epidemic of the new coronavirus has brought a huge threat to people's life safety. The reason for the rapid outbreak of the new coronavirus is the lack of effective detection methods; the current standard detection method recognized by the world is PCR detection, but this method has certain defects, the detection cycle is long, and it takes about two to three ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/24C12M1/34C12Q1/6844
CPCC12Q1/6844C12Q2521/327C12Q2525/161C12Q2531/113C12N9/22C12Q1/6806B01L7/52B01L3/5082B01L3/50825B01L2300/0854B01L2300/161B01L2400/0409C12Q2521/507C12Q2523/32C12Q2527/101C12Q2531/10C12N2310/20B01L3/5021B01L2200/141B01L2300/043B01L2300/0832B01L2300/12B01L2300/165C12N15/11C12N2800/80C12Q1/6876
Inventor 胡飞刘艳飞彭年才
Owner XI AN JIAOTONG UNIV
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