CN212504862U

CN212504862U - Totally enclosed sample nucleic acid extraction purification device

Info

Publication number: CN212504862U
Application number: CN202020895891.2U
Authority: CN
Inventors: 李康
Original assignee: Yantai Ausbon Biological Research And Development Co ltd
Current assignee: Yantai Ausbon Biological Research And Development Co ltd
Priority date: 2020-05-25
Filing date: 2020-05-25
Publication date: 2021-02-09
Anticipated expiration: 2030-05-25

Abstract

The utility model discloses a totally enclosed sample nucleic acid extraction and purification device, which comprises a closed sample cracking container and a sample purification container; one surface of the sample cracking container is inwards provided with a cracking liquid chamber, a sample chamber, an extracting liquid chamber and a combining liquid chamber which are sequentially communicated through a gas channel; a filter I, a filter II and a filter III are respectively arranged in the lysate chamber, the sample chamber and the binding solution chamber; the top end of the sample chamber is provided with a sample inlet extending out of the sample cracking container; the top ends of the lysate cavity and the bonding solution cavity are both sealed by tinfoil; the top end of the extracting solution chamber is provided with a pierceable heat sealing film. The utility model discloses an all operations are all gone on in airtight environment, have avoided the pollution problem of sample and nucleic acid extract, when handling the sample that has the infection risk, have fine guard action to operating personnel and laboratory environment.

Description

Totally enclosed sample nucleic acid extraction purification device

Technical Field

The utility model belongs to the technical field of nucleic acid draws, especially, relate to a totally enclosed sample nucleic acid draws purification device.

Background

When a solid biological detection material (including a sample cotton swab, a cast-off cell adhesive device, a cut clothes spot, a rope and the like) is used for nucleic acid extraction, one inevitable process is that the lysis, centrifugation and carrier removal are carried out, and then a lysate is absorbed for subsequent treatment. The currently mainstream technology of the treatment process is to operate by using a centrifugal sleeve, and both manual operation and automatic operation need to separate an inner tube filled with a sample carrier and an outer tube filled with a centrifugally recovered product so as to take out a lysate. While a bidirectional tube suitable for manual operation is disclosed in patent No. CN208260788U, the centrifuge can be transferred to another tube after inversion, and the tube is not removed, but an automatic procedure cannot be realized, and the lysate is taken out by manually unscrewing the cover.

At present, the magnetic bead method sample nucleic acid extraction and purification technology is mainly divided into the following two types:

the other method is a liquid transfer method, after the cracking product is combined with the magnetic bead, the magnetic bead is adsorbed by a magnetic device, the magnetic bead is left in situ, the waste liquid is transferred and discarded by a liquid processing channel, then new washing liquid is added to wash the magnetic bead, the waste liquid is discarded, finally eluent is added to elute the nucleic acid combined with the magnetic bead, and the eluted product is transferred away to be used as a nucleic acid sample. The method is long in time consumption, high in material consumption, and high in heating requirement in the final elution process, and has the risks of liquid evaporation and aerosol generation.

The other method is a magnetic bead transferring method, wherein magnetic beads are transferred between different washing solutions and different eluents by matching a magnetic rod with a magnetic sleeve. Although the method is short in time consumption, the elution and heating cannot be closed, cross contamination is easily caused by shaking or rotating and uniformly mixing the magnetic sleeve, and the risk of aerosol generation is caused.

SUMMERY OF THE UTILITY MODEL

The utility model discloses to the not enough of above-mentioned prior art existence, provide a labour saving and time saving, avoid polluting and be fit for the totally enclosed sample nucleic acid extraction purification device of automatic batch processing sample.

The utility model provides an above-mentioned technical problem's technical scheme as follows: a totally enclosed sample nucleic acid extraction and purification device is characterized by comprising a closed sample cracking container and a sample purification container;

one surface of the sample cracking container is inwards provided with a cracking liquid chamber, a sample chamber, an extracting liquid chamber and a combining liquid chamber in parallel; the upper parts of the lysate chamber, the sample chamber, the extracting solution chamber and the bonding solution chamber are sequentially communicated through a gas channel; a filter I, a filter II and a filter III are respectively arranged in the lysate chamber, the sample chamber and the binding solution chamber, and the filter I, the filter II and the filter III respectively divide the lysate chamber, the sample chamber and the binding solution chamber into an upper section and a lower section; the lower section of the lysate chamber is communicated to the upper section of the sample chamber through a liquid channel I; the lower section of the sample chamber is communicated to the extracting solution chamber through a liquid channel II; the lower section of the combination liquid chamber is communicated to the extracting liquid chamber through a liquid channel III; the top end of the sample chamber is provided with a sample inlet extending out of the sample cracking container, the outer surface of the sample inlet is provided with external threads, and the sample inlet is in threaded connection with a pipe cover of which the inner surface is provided with internal threads; the top ends of the lysate cavity and the bonding solution cavity are both sealed by tinfoil; the top end of the extracting solution cavity is provided with a pierceable heat sealing film;

one surface of the sample purification container is inwards provided with a mixing chamber, a first washing chamber, a second washing chamber and an elution chamber in parallel, wherein the upper parts of the mixing chamber, the first washing chamber, the second washing chamber and the elution chamber are communicated; the top ends of the mixing chamber, the first washing chamber, the second washing chamber and the elution chamber are all pierceable heat-sealing films; magnetic beads which can move under the action of external magnetic force are arranged in the mixing chamber.

Furthermore, the pore diameter of the filter I is 1-20 μm; the pore diameter of the filter II is 0.22-0.85 μm; the pore diameter of the filter III is 0.22-0.85 μm.

The beneficial effect of adopting the above further scheme is that the pore size of the filter is controlled, thereby avoiding blocking each liquid channel.

Furthermore, the pipe diameters of the liquid channels I, II and III are 0.5-3 mm.

The beneficial effect who adopts above-mentioned further scheme is that, through the gas passage intercommunication between each cavity, guarantee atmospheric pressure balance to rely on centrifugal force, make the liquid in each cavity pass through liquid passage and shift.

Furthermore, the lysate chamber is a plurality of.

Further, the sample cracking container and the sample purifying container are both made of transparent materials; preferably, the sample lysis container and the sample purification container are made into a sample lysis reagent card and a sample purification reagent card.

The utility model discloses a characteristics and beneficial effect are:

in the utility model, the tube cover is only required to be opened when the sample is added, thus ensuring the sealing property in the processing process; separating the sample and the lysis reagent, and mixing only in a centrifugal state; the processes of heating cracking and heating elution are closed operations, so that the problem of evaporation is avoided. And the magnetic bead principle is combined again, consumable (magnetic sleeve) and liquid transfer sample adding tips are saved, the whole treatment process only needs two sample adding tips, and compared with the prior art, 2/3 sample adding tips are saved. The utility model discloses an all operations are all gone on in airtight environment, have avoided the pollution problem of sample and nucleic acid extract, when handling the sample that has the infection risk, have fine guard action to operating personnel and laboratory environment. The device and the method of the utility model have the advantages of fast processing speed, higher safety, low cost and suitability for manual and automatic operation.

Drawings

FIG. 1 is a schematic diagram of a sample lysis vessel;

FIG. 2 is a schematic diagram of a sample purification vessel;

in the figure, 1, a sample lysis vessel; 1-1, a lysate chamber; 1-2, a sample chamber; 1-3, an extracting solution chamber; 1-4, a binding liquid chamber; 1-5, gas channels; 1-6, a filter I; 1-7, filter II; 1-8, filter III; 1-9, a liquid channel I; 1-10, liquid channel II; 1-11, liquid channel III; 1-12, a tube cover;

2. a sample purification vessel; 2-1, a mixing chamber; 2-2, a first washing chamber; 2-3, a second washing chamber; 2-4, an elution chamber; 2-5, magnetic beads.

Detailed Description

The principles and features of the present invention are described below in conjunction with the following drawings, the examples given are only intended to illustrate the present invention and are not intended to limit the scope of the present invention.

A totally enclosed sample nucleic acid extraction and purification device is characterized by comprising a closed sample cracking container 1 and a sample purification container 2;

one surface of the sample cracking container is inwards provided with a cracking liquid chamber 1-1, a sample chamber 1-2, an extracting liquid chamber 1-3 and a binding liquid chamber 1-4 in parallel; the upper parts of the lysate chamber, the sample chamber, the extracting solution chamber and the bonding solution chamber are sequentially communicated through gas channels 1-5; the filter I, the filter II and the filter III divide the lysis solution chamber, the sample chamber and the binding solution chamber into an upper section and a lower section respectively; the lower section of the lysate chamber is communicated to the upper section of the sample chamber through liquid channels I1-9; the lower section of the sample chamber is communicated to the extracting solution chamber through liquid channels II 1-10; the lower section of the combination liquid chamber is communicated to the extracting liquid chamber through liquid channels III 1-11; the top end of the sample chamber is provided with a sample inlet extending out of the sample cracking container, the outer surface of the sample inlet is provided with external threads, and the sample inlet is in threaded connection with a pipe cover 1-12, the inner surface of which is provided with internal threads; the top ends of the lysate cavity and the bonding solution cavity are both sealed by tinfoil; the top end of the extracting solution cavity is provided with a pierceable heat sealing film;

one surface of the sample purification container is inwards provided with a mixing chamber 2-1, a first washing chamber 2-2, a second washing chamber 2-3 and an elution chamber 2-4 which are communicated with each other at the upper parts side by side; the top ends of the mixing chamber, the first washing chamber, the second washing chamber and the elution chamber are all pierceable heat-sealing films; magnetic beads 2-5 capable of moving under the action of external magnetic force are arranged in the mixing chamber;

further, the pore diameter of the filter I is 5 mu m; the pore diameter of the filter II is 0.65 mu m; the pore size of the filter III is 0.65 mu m;

furthermore, the pipe diameters of the liquid channels I, II and III are 2 mm.

A method for extracting and purifying nucleic acid by using the totally enclosed sample nucleic acid extraction and purification device comprises the following steps:

(1) mixing materials: adding lysis solution into the lysis solution chamber; adding binding liquid to the binding liquid chamber; the solid biological detection material to be extracted is plugged into the sample chamber, and the tube cover is covered; placing the sample cracking container in a centrifuge, carrying out low-speed centrifugation at the rotating speed of 300r/min, allowing the lysate in the lysate chamber to enter the sample chamber through a liquid channel I under the action of centrifugal force until the lysate completely covers the solid biological detection material, and stopping centrifugation;

(2) cracking: heating the sample cracking container to 65 ℃, and completing cracking of the solid biological detection material; placing the sample cracking container in a centrifuge, carrying out high-speed centrifugation at the rotating speed of 4000r/min, wherein under the action of centrifugal force, the nucleic acid extracting solution in the sample chamber enters an extracting solution chamber through a liquid channel II, and meanwhile, the liquid in the combined liquid chamber also enters the extracting solution chamber through a liquid channel III;

(3) and (3) purification: taking out the nucleic acid extracting solution in the extracting solution cavity to a sample purification container by a liquid transferring device through a pierceable heat sealing film at the top end of the extracting solution cavity; in the mixing chamber, combining the nucleic acid extracting solution with magnetic beads, adsorbing the magnetic beads to move to a first washing chamber and a second washing chamber along the inner wall under the action of external magnetic force, finally entering an elution chamber, and heating and eluting at 40 ℃; thus obtaining the purified nucleic acid extracting solution.

Further, when the magnetic beads move to the first washing chamber and the second washing chamber respectively, the external magnetic force is removed, and the magnetic beads are vibrated and suspended in the first washing chamber and the second washing chamber for 3 times respectively.

The above description is only for the preferred embodiment of the present invention, and is not intended to limit the present invention, and any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention should be included within the protection scope of the present invention.

Claims

1. A totally enclosed sample nucleic acid extraction and purification device is characterized by comprising a closed sample cracking container and a sample purification container;

2. The totally enclosed sample nucleic acid extraction and purification device according to claim 1, wherein the pore size of the filter i is 1 to 20 μm; the pore diameter of the filter II is 0.22-0.85 μm; the pore diameter of the filter III is 0.22-0.85 μm.

3. The totally enclosed sample nucleic acid extraction and purification device according to claim 1, wherein the tube diameters of the liquid channels I, II are 0.5-3 mm.

4. The totally enclosed sample nucleic acid extraction and purification device according to claim 1, wherein the lysate chamber is multiple.