Front-end treatment device for tumor sample pretreatment and RNA (Ribonucleic Acid) extraction

Abstract

The utility model discloses a front-end treatment device for tumor sample pretreatment and RNA (Ribonucleic Acid) extraction. The front-end treatment device comprises a centrifugal tube and an extraction component, the extraction assembly comprises an extraction pipe group and a liquid injector; the extraction tube group comprises a first tube body, a second tube body and a third tube body, pores are formed in the bottom of the first tube body and the bottom of the second tube body, and a first filter membrane of 20-40 microns is fixedly coated at the bottom of the first tube body; the bottom of the second tube body is coated and fixed with a second filter membrane of 100-300nm; the first pipe body is arranged on the inner side of the second pipe body in a hollow embedded mode, and a first sealing ring is arranged between the upper portion of the outer side of the first pipe body and the upper portion of the inner side of the second pipe body. According to the front-end treatment device for tumor sample pretreatment and RNA extraction, the centrifugal tube is adopted for dewaxing and enzymolysis of a tissue sample, then step-by-step splitting decomposition and filtration are conducted through the extraction assembly, operation is convenient, and the RNA extraction efficiency can be improved.

Description

Technical Field

[0001] The utility model specifically relates to a front-end processing device for tumor sample preprocessing and RNA extraction, and belongs to the technical field of RNA extraction. Background Art

[0002] Traditional methods for detecting gene mutations, for example, mainly detect gene mutations based on the comparison results of DNA sequencing data of the sample to be tested; RNA editing occurs after DNA transcription, and RNA can then be translated into protein, and not all gene mutations are transcribed, so there is a certain difference between the gene mutations detected based on DNA sequencing data and the gene mutations actually expressed; in addition, since RNA is more difficult to preserve than DNA, and there are a variety of alternative splicing in RNA, it is difficult to accurately detect somatic gene mutations at the RNA level. In existing technologies, tissue samples are usually preserved by formalin fixation and paraffin embedding; in this way, the morphological characteristics of the original tissue sample are completely preserved; with the development of molecular biology technology, more and more molecular diagnostic services use formalin fixed paraffin embedded (FFPE) tissue samples; FFPE samples contain a lot of clinically relevant information, but it is very difficult to extract RNA from tissue samples. Utility Model Content

[0003] In order to solve the above problems, the utility model proposes a front-end processing device for tumor sample pretreatment and RNA extraction, which is easy to operate and can simplify the RNA extraction efficiency by dewaxing, enzymatic hydrolysis and lysis of tissue samples.

[0004] The utility model discloses a front-end processing device for tumor sample pretreatment and RNA extraction, comprising a centrifuge tube and an extraction component; the extraction component comprises an extraction tube set and a liquid injector; the extraction tube set comprises

[0005] A first tube body, wherein a hole is opened at the bottom of the first tube body and a first filter membrane of 20-40 μm is fixedly coated thereon;

[0006] A second tube body, wherein the bottom of the second tube body is provided with a hole and is covered and fixed with a second filter membrane of 100-300 nm; the first tube body is hollow and embedded in the inner side of the second tube body, and a first sealing ring is arranged between the upper part of the outer side of the first tube body and the upper part of the inner side of the second tube body;

[0007] A third tube body, wherein the second tube body is hollow and embedded inside the third tube body, and a second sealing ring is arranged between the upper outer portion of the second tube body and the upper inner portion of the third tube body;

[0008] The first tube body is provided with a first liquid injection port above the second tube body, and the second tube body is provided with a second liquid injection port above the third tube body; rubber plugs are provided in the first liquid injection port and the second liquid injection port.

[0009] The injectors include a dewaxing injector, an enzymolysis injector and a lysis solution injector.

[0010] Furthermore, the top of the first tube body is embedded in the sealing plug.

[0011] Furthermore, the centrifuge tube, extraction assembly and injector are arranged in the same packaging box.

[0012] Furthermore, it also includes an oscillation device and a centrifugal device.

[0013] The working process of the front-end processing device is as follows: first, cut multiple pieces of paraffin-embedded tumor samples, send them into a centrifuge tube, and inject D-limonene dewaxing solution into the centrifuge tube through a dewaxing injector, first oscillate for 20 seconds through an oscillating device, and then centrifuge for 2 minutes at room temperature through a centrifuge device; after the centrifuge tube is allowed to stand, the supernatant in the centrifuge tube is extracted by filtration; then, the protease solution is injected into the centrifuge tube through an enzymatic hydrolysis injector, first enzymatically hydrolyze at room temperature for 30 minutes, and then the centrifuge tube is sent into the centrifuge device, and the centrifugal temperature is controlled at 50°C and centrifuged for 10 minutes; after the centrifugation is completed, the sealing plug is pulled out, the mixed solution in the centrifuge tube is put into the first tube body, the rubber plug is inserted through the lysate injector, and the lysate is injected The liquid injector injects the lysate into the first tube body through the first liquid injection port, and controls the centrifugal temperature to 50°C for centrifugal extraction. When the liquid in the first tube body penetrates into the second tube body through the first filter membrane, the rubber plug is inserted through the lysate injector, and the lysate injector injects the lysate into the second tube body through the second liquid injection port, and controls the centrifugal temperature to 50°C for centrifugal extraction. When the liquid in the second tube body penetrates into the third tube body through the first filter membrane, the centrifugation is completed; the first tube body and the second tube body are discarded; the liquid obtained in the third tube body is used as the front-end processing liquid for RNA extraction to complete the front-end processing of RNA extraction; the front-end processing liquid is analyzed after being adsorbed by the later adsorption process, or the RNA is analyzed by nanopore sequencing.

[0014] Compared with the prior art, the front-end processing device for tumor sample pretreatment and RNA extraction of the utility model uses a centrifuge tube to dewax and enzymatically hydrolyze tissue samples, and then performs step-by-step lysis and filtration through an extraction component, which is easy to operate and can simplify RNA extraction efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 It is a schematic diagram of the overall structure of the utility model.

[0016] Figure 2 It is a schematic diagram of the matching structure of the centrifuge tube and the liquid injector of the utility model.

[0017] Figure 3 It is a schematic diagram of the matching structure of the extraction component and the liquid injector of the utility model. DETAILED DESCRIPTION

[0018] Example:

[0019] like Figures 1 to 3 The front-end processing device for tumor sample pretreatment and RNA extraction shown in the figure includes a centrifuge tube 1 and an extraction component; the extraction component includes an extraction tube group and an injector; the extraction tube group includes

[0020] A first tube body 2, wherein the first tube body 2 has a hole at the bottom and is covered and fixed with a first filter membrane 3 of 20-40 μm;

[0021] A second tube body 4, wherein the second tube body 4 has a hole at the bottom and is covered and fixed with a second filter membrane 5 of 100-300 nm; the first tube body 2 is hollow and embedded inside the second tube body 4, and a first sealing ring 6 is arranged between the upper outer part of the first tube body 2 and the upper inner part of the second tube body 4;

[0022] A third tube body 7, wherein the second tube body 4 is hollow and embedded inside the third tube body 7, and a second sealing ring 8 is provided between the upper outer portion of the second tube body 4 and the upper inner portion of the third tube body 7;

[0023] The first tube body 2 is provided with a first liquid injection port 9 above the second tube body 4 , and the second tube body 4 is provided with a second liquid injection port 10 above the third tube body 7 ; rubber plugs 11 are provided in the first liquid injection port 9 and the second liquid injection port 10 .

[0024] The injectors include a dewaxing injector 12 , an enzymolysis injector 13 and a lysis solution injector 14 .

[0025] The top of the first tube body 2 is embedded in the sealing plug 15. The centrifuge tube 1, the extraction assembly and the injector are arranged in the same packaging box. An oscillating device and a centrifugal device are also included.

[0026] The working process of the front-end processing device is as follows: first, cut a plurality of paraffin-embedded tumor samples, send them into the centrifuge tube 1, and inject D-limonene dewaxing liquid into the centrifuge tube 1 through the dewaxing liquid injector 12, first oscillate for 20 seconds through the oscillation device, and then centrifuge for 2 minutes at room temperature through the centrifuge device; after the centrifuge tube 1 is allowed to stand, the supernatant in the centrifuge tube 1 is extracted by filtration; then, the protease solution is injected into the centrifuge tube 1 through the enzymatic liquid injector 13, first enzymolyze at room temperature for 30 minutes, and then the centrifuge tube 1 is sent into the centrifuge device, and the centrifugal temperature is controlled at 50°C and centrifuged for 10 minutes; after the centrifugation is completed, the sealing plug 15 is pulled out, and the mixed liquid in the centrifuge tube 1 is put into the first tube body 2, and the rubber plug 11 is inserted through the lysate liquid injector 14, and the lysate liquid injector is inserted into the rubber plug 11. 14 injects the lysate into the first tube body 2 through the first injection port 9, and controls the centrifugal temperature to 50°C for the centrifugal extraction component. When the liquid in the first tube body 2 penetrates into the second tube body 4 through the first filter membrane 3, the rubber plug 11 is inserted through the lysate injector 14. The lysate injector 14 injects the lysate into the second tube body 4 through the second injection port 10, and controls the centrifugal temperature to 50°C for the centrifugal extraction component. When the liquid in the second tube body 4 penetrates into the third tube body 7 through the first filter membrane 3, the centrifugation is completed; the first tube body 2 and the second tube body 4 are discarded; the liquid obtained in the third tube body 7 is used as the front-end processing liquid for RNA extraction to complete the front-end processing of RNA extraction; the front-end processing liquid is analyzed after being adsorbed by the later adsorption process, or the RNA is analyzed by nanopore sequencing.

[0027] The above embodiments are only preferred implementations of the present invention, so any equivalent changes or modifications made according to the structures, features and principles described in the scope of application of the present invention are included in the scope of application of the present invention.

Claims

1. A front-end processing device for tumor sample pretreatment and RNA extraction, characterized in that: It includes a centrifuge tube and an extraction component; the extraction component includes an extraction tube set and a liquid injector; the extraction tube set includes A first tube body, wherein a hole is opened at the bottom of the first tube body and a first filter membrane of 20-40 μm is fixedly coated thereon; A second tube body, wherein the bottom of the second tube body is provided with a hole and is covered and fixed with a second filter membrane of 100-300 nm; the first tube body is hollow and embedded in the inner side of the second tube body, and a first sealing ring is arranged between the upper part of the outer side of the first tube body and the upper part of the inner side of the second tube body; A third tube body, wherein the second tube body is hollow and embedded inside the third tube body, and a second sealing ring is arranged between the upper outer portion of the second tube body and the upper inner portion of the third tube body; The first tube body is provided with a first liquid injection port above the second tube body, and the second tube body is provided with a second liquid injection port above the third tube body; rubber plugs are provided in the first liquid injection port and the second liquid injection port; The injectors include a dewaxing injector, an enzymolysis injector and a lysis solution injector.

2. The front-end processing device for tumor sample pretreatment and RNA extraction according to claim 1, characterized in that: The top of the first tube is embedded in the sealing plug.

3. The front-end processing device for tumor sample pretreatment and RNA extraction according to claim 1, characterized in that: The centrifuge tube, the extraction component and the liquid injector are arranged in the same packaging box.

4. The front-end processing device for tumor sample pretreatment and RNA extraction according to claim 1, characterized in that: Also includes shaking equipment and centrifugal equipment.

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