WO2020073829A1

WO2020073829A1 - Separation device and use thereof, detection system, electrochemical detection system and cell sorting system

Info

Publication number: WO2020073829A1
Application number: PCT/CN2019/108677
Authority: WO
Inventors: 范凌云
Original assignee: 广东菲鹏生物有限公司
Priority date: 2018-10-08
Filing date: 2019-09-27
Publication date: 2020-04-16
Also published as: CN109486651A; CN109486651B

Abstract

The present application discloses a separation device and use thereof, a detection system, an electrochemical detection system and a cell sorting system. The separation device comprises a cylinder body, a diaphragm assembly and a sliding member, the cylinder body having a first end and a second end opposite each other; the diaphragm assembly comprises a plurality of partition diaphragms and a plurality of blocking diaphragms, the plurality of partition diaphragms being accommodated in the cylinder body and being sealingly connected to the cylinder body, the plurality of partition diaphragms being arranged at intervals from the second end to the first end to divide the cylinder body into an end chamber and N functional chambers, the end chamber being close to the second end, side walls of the N functional chambers being provided with liquid discharge holes, each liquid discharge hole being blocked by the blocking diaphragm; and the sliding member is provided close to the first end and is slidable toward a direction close to the second end. The separation device above causes less contamination to a sample to be tested.

Description

Separation device and its application, detection system, electrochemical detection system and cell sorting system

Cross-reference of related applications

This application requires the priority of the Chinese patent application filed on October 8, 2018 with the application number 201811169486.6, entitled "Separation device and its application, detection system, electrochemical detection system and cell sorting system", The entire contents are incorporated by reference in this application.

Technical field

The present application relates to the technical field of separation devices, in particular to a separation device and its application, detection system, electrochemical detection system and cell sorting system.

Background technique

The magnetic bead separation technology mainly uses magnetic beads to specifically adsorb the material to be separated to separate the material to be separated. Because of its strong specificity, safety and non-toxicity, it is widely used in nucleic acid extraction and chemiluminescence detection. At present, during the use of most magnetic bead separation devices, the sample to be tested needs to be loaded into a centrifuge tube and then placed in the magnetic bead separation device for separation operation. This method requires the process of repeatedly opening and adding the liquid, mixing and mixing the cover, and opening and discarding the liquid. Repeated opening of the cover is likely to cause contamination of the sample to be tested.

Application content

Based on this, it is necessary to provide a separation device that can at least achieve less contamination of the sample to be tested. In addition, it provides an application of separation device, detection system, electrochemical detection system and cell sorting system.

A separation device, including:

The barrel has opposite first and second ends;

The diaphragm assembly includes a plurality of separation membranes and a plurality of sealing membranes. The plurality of separation membranes are all accommodated in the cylinder body and are all sealedly connected to the cylinder body. The plurality of separation membranes extend from the second end Spaced toward the first end to divide the barrel into an end chamber and N functional chambers, the end chamber is close to the second end, the sidewalls of the N functional chambers There are drain holes on the openings, and each of the drain holes is blocked with the blocking film; and

The slider is provided near the first end and can slide toward the second end.

In the above separation device, the cylinder body is divided into an end chamber and N functional chambers by a plurality of separation membranes, and the plurality of separation membranes are all sealedly connected to the cylinder body, and the side walls of the N functional chambers are all provided with Drainage holes, each of which is sealed with a blocking membrane, can be placed in the functional chamber closest to the first end to be configured to adsorb the target material adsorbent, can be placed in other functional chambers or the reaction liquid Cleaning fluid, etc .; because the slider is located near the first end and can slide toward the second end, when the target substance needs to be separated, the slider can be accommodated in the functional chamber closest to the first end and can be slid An adsorption member is provided on the side away from the first end, and the sample to be separated is added to adsorb the target substance on the adsorption member, and the sliding member is slid toward the second end, so that the sealing membrane and the separation membrane are broken and carried The adsorbent that adsorbs the target substance enters other functional chambers for reaction or cleaning, and finally enters the end chamber to obtain the pure target substance; when the target substance needs to be detected, the pure target substance can be removed from the end cavity It can be detected in the middle-inflow detection device, or it can be directly detected in the end chamber, or a reaction reagent or detection reagent can be placed in the end chamber to make the target substance react with the reaction reagent or detection reagent to detect the target substance. The above separation device is simple, and the target substance can be separated and detected without repeatedly opening the cover, and the operation is convenient, and the sample to be tested is less polluted.

Optionally, the separating device further includes a pushing member, and the pushing member is detachably connected with the sliding member.

Optionally, it further includes an adsorption member, which is accommodated in the functional chamber closest to the first end, the pushing member is at least partially magnetic, and when the pushing member is connected to the sliding member The pushing member can adsorb the adsorption member and the adsorption member can adsorb on the sliding member.

Optionally, the separation device further includes a magnetic field generating mechanism configured to provide a magnetic field to the adsorption member, and the adsorption member can move under the action of the magnetic field.

Optionally, the adsorption member is a magnetic bead.

Optionally, the magnetic beads are superparamagnetic silicon oxide nano magnetic beads or carboxylated magnetic beads.

Optionally, an adsorption member is further included, and the adsorption member is a chip. The adsorption member is accommodated in the functional chamber closest to the first end and fixed to the sliding member.

Optionally, the tensile strength of the separation film is greater than the tensile strength of the blocking film, and when the sliding member slides in a direction close to the second end, the functional cavity where the sliding member is located can be made The pressure in the chamber is increased, so that the blocking film and the separation film of the functional chamber where the slider is located can be sequentially broken.

Optionally, a waste liquid tank is further included, and the waste liquid tank is in communication with the drain hole.

Optionally, the waste liquid tank is a ring-shaped hollow structure, the cylinder is at least partially contained in the waste liquid tank, and is sealedly connected to the waste liquid tank;

Or, the waste liquid tank is cylindrical, and the outer wall of the waste liquid tank is fixed to the outer wall of the barrel body.

Optionally, a side wall of the adsorption chamber is further provided with an air pressure balancing hole communicating with the waste liquid tank, and the air pressure balancing hole is provided near the first end.

Optionally, the cylinder body is provided with a sample port, the sample port is located on the cavity wall of the functional chamber closest to the first end, and the separation device further includes a configuration configured to seal the sample port Sample port seals;

The cylinder body is provided with a sample outlet, the sample outlet is communicated with the end chamber, and the separation device further includes a stopper configured to seal the sample outlet.

Optionally, the barrel includes a barrel body, a first side plate and a second side plate;

The first end and the second end are provided with a first opening and a second opening, the first opening is opposite to the second opening, and the first side plate and the second side plate respectively shield the The first opening and the second opening are both hermetically connected to the barrel body.

Optionally, the sliding member includes a sliding part and a mounting part fixedly connected to the sliding part;

The sliding portion is provided near the first end and can slide toward the second end; the sliding portion is housed in the functional chamber near the first end and is connected to the body of the cylinder Sealed connection on the inner wall;

The mounting portion is provided near the first end and can slide toward the second end; the mounting portion is partially accommodated in the functional chamber near the first end and can seal the cylinder.

The application of the above-mentioned separation device in nucleic acid extraction, chemiluminescence detection or cell sorting.

A detection system includes the separation device described above.

Optionally, the detection system is a PCR (Polymerase Chain Reaction) detection system, the end chamber is a detection chamber, and the functional chamber closest to the first end is a sample chamber , At least one of the remaining functional chambers is a cleaning chamber;

Or, the detection system is a chemiluminescence detection system, the end chamber is a detection chamber, the functional chamber closest to the first end is a sample chamber, and at least one of the remaining functional chambers For the reaction chamber.

An electrochemical detection system is characterized by including the above-mentioned separation device.

Optionally, the end chamber is a detection chamber, the functional chamber closest to the first end is a sample chamber, and at least one of the remaining functional chambers is a cleaning chamber.

A cell sorting system is characterized by including the above-mentioned separation device.

BRIEF DESCRIPTION

1 is a schematic structural diagram of a separation device according to an embodiment;

2 is a schematic structural view of the separation device shown in FIG. 1 after omitting the waste liquid tank;

FIG. 3 is a schematic structural view of a sliding member and a pushing member in the separation device shown in FIG. 1;

4 is a schematic structural view of a separation device in a chemiluminescence detection system after a waste liquid tank is omitted.

Icons: 10-separation device; 100-barrel; 110-barrel body; 111-first end; 113-second end; 120-first side plate; 130-second side plate; 140-end chamber 144-air pressure balance hole; 150-function chamber; 160-drain hole; 170-sample inlet; 190-sample outlet; 200-diaphragm assembly; 210-partition membrane; 220-blocking membrane; 300-slide Parts; 310-sliding part; 320-mounting part; 322-accommodating groove; 400-pushing part; 500-seal; 600-waste liquid tank.

detailed description

In order to facilitate understanding of the application, the application will be described more fully below with reference to related drawings. The drawings show preferred embodiments of the present application. However, this application can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present application more thorough and comprehensive.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the technical field of the present application. The terminology used in the specification of the present application herein is for the purpose of describing specific embodiments only, and is not intended to limit the present application.

As shown in FIG. 1, the detection system of an embodiment includes a separation device 10 and a detection device (not shown). The separation device 10 is configured to separate the target substance in the sample to be measured. The detection device communicates with the separation device 10 to perform qualitative or quantitative detection of the target substance in the sample to be tested.

Optionally, the sample to be tested is blood or saliva. The target substances are nucleic acids and target cells. It should be noted that the sample to be tested is not limited to the above-mentioned pointed sample, and the target substance is not limited to the above-mentioned pointed substance, and can be set according to the actual situation.

The separation device 10 includes a cylinder 100, a diaphragm assembly 200, a sliding member 300, a pushing member 400, and a waste tank 600. In the illustrated embodiment, the target substance is a nucleic acid. The separation device 10 is configured for nucleic acid extraction.

Please refer to FIG. 2 together. The barrel 100 is the outer shell of the separating device 10. The barrel 100 includes a barrel body 110, a first side plate 120 and a second side plate 130. The barrel body 110 has a first end 111 and a second end 113 opposite to each other. The first end 111 and the second end 113 define a first opening (not shown) and a second opening (not shown). The first opening is opposite to the second opening. The first side plate 120 and the second side plate 130 respectively cover the first opening and the second opening, and are both sealedly connected to the barrel body 110.

It should be noted that the cylindrical body 110, the second side plate 130, and the first side plate 120 may be an integrally formed structure, or may be other fixing methods, such as welding.

In the illustrated embodiment, the cylindrical body 110 is cylindrical. Both the first side plate 120 and the second side plate 130 are disc-shaped. The first side plate 120 is opposite to and parallel to the second side plate 130. The outer diameter of the first side plate 120 is approximately the same as the inner diameter of the first opening, so that the first side plate 120 just covers the first opening; The second side plate 130 just covers the second opening.

It should be noted that the cylindrical body 110 is not limited to the above-mentioned pointed shape, and may be other shapes, such as a square cylindrical shape. Correspondingly, the first side plate 120 and the second side plate 130 are not limited to the above-mentioned shapes, and may be other shapes, such as a square shape. It can be set according to the actual situation, as long as the first side plate 120 and the second side plate 130 cover the first opening and the second opening, respectively, and both are fixedly connected and hermetically connected to the barrel body 110.

The diaphragm assembly 200 includes a plurality of separation films 210 and a plurality of blocking films 220. The plurality of separation films 210 are all contained in the cylinder 100 and are all sealedly connected to the cylinder 100. The separation membranes 210 are arranged at intervals from the second end 113 to the first end 111 to divide the cylinder 100 into an end chamber 140 and N functional chambers 150. The end chamber 140 is disposed near the second end 113. The side walls of the N functional chambers 150 are all provided with drainage holes 160, and each drainage hole 160 is sealed with a blocking film 220.

Optionally, the detection system is a PCR (Polymerase Chain Reaction) detection system. The end chamber 140 is a detection chamber, the functional chamber 150 closest to the first end is a sample chamber, and at least one of the remaining functional chambers 150 is a cleaning chamber.

In the illustrated embodiment, there are three separation films 210. Each separation film 210 has a disc shape, and is arranged parallel to the first side plate 120. The three separation films 210 are arranged at intervals from the second end 113 to the first end 111 to divide the cylinder 100 into an end chamber 140 and three functional chambers 150. The end chamber 140 is configured to contain the eluent. The functional chamber 150 closest to the first end 111 can accommodate the sample to be tested. The remaining two functional chambers 150 are configured to contain cleaning fluid to clean the target substance. The side walls of the three functional chambers 150 are all provided with drainage holes 160. There are three blocking films 220. The three blocking films 220 respectively block the drainage holes 160 of the three functional chambers 150.

It should be noted that the cleaning liquids in the two functional chambers 150 configured to receive the cleaning liquid may be the same cleaning liquid or different cleaning liquids, and can be set according to actual conditions.

Optionally, the separation film 210 is an acrylic film, a nylon film, a Teflon film, a polyethylene film, a polypropylene film, or a polyolefin film. It should be noted that the separation film 210 is not limited to the above-mentioned film, and any film commonly used in the art may be used as the separation film 210.

Optionally, the blocking film 220 is an acrylic film, nylon film, Teflon film, polyethylene film, polypropylene film or polyolefin film. It should be noted that the blocking film 220 is not limited to the above-mentioned film, and any film commonly used in the art may be used as the blocking film 220.

Further, the cylinder 100 is provided with a sample port 170. The sample port 170 is located on the wall of the functional chamber 150 closest to the first end 111. The sample to be tested can be added into the barrel 100 from the sample port 170. Furthermore, the separation device 10 further includes a sealing member 500 configured to seal the sampling port 170. In the illustrated embodiment, the sample port 170 is spaced from and opposite to the drain hole 160. The sealing member 500 is a plug and can be inserted into the sample port 170 to seal the sample port 170. It should be noted that the sealer 500 is not limited to a plug, but may also be a lid body, which is provided on the sampling port 170 to seal the sampling port 170.

The cylinder 100 is provided with a sample outlet 190. The sample outlet 190 communicates with the end chamber 140. Further, the separation device 10 further includes a stopper (not shown) configured to seal the sample outlet 190. In the illustrated embodiment, the sample outlet 190 is opened in the second side plate 130 and penetrates the second side plate 130. The occluder may be a plug so as to be able to be inserted into the sample outlet 190 and seal the sample outlet 190. It should be noted that the stopper may also be a cover so as to cover the sample outlet 190 and seal the sample outlet 190.

Optionally, the sample port 170 and the sample port 190 can be set as a circular through hole or a square through hole, as long as the sample to be tested can be added to the barrel 100 from the sample port 170, and the tested sample can be taken out The sample port 190 only needs to exit the barrel 100, and will not be repeated here.

The slider 300 is disposed near the first end 111 and can slide toward the second end 113. Further, the tensile strength of the separation film 210 is greater than the tensile strength of the blocking film 220. When the slider 300 slides toward the second end 113, the pressure in the functional chamber 150 where the slider 300 is located can be increased, and the sealing film 220 and the separation membrane 210 of the functional chamber 150 where the slider 300 is located can be increased. Able to break in sequence. Furthermore, the slider 300 is at least partially accommodated in the functional chamber 150 closest to the first end 111 and seals the functional chamber 150 closest to the first end 111.

Please refer to FIG. 3 together. The sliding member 300 includes a sliding part 310 and a mounting part 320 fixed to the sliding part 310.

The sliding portion 310 is provided near the first end 111 and can slide toward the second end 113. Further, the sliding portion 310 is accommodated in the functional chamber 150 closest to the first end 111 and is sealingly connected to the inner wall of the barrel body 110. In the illustrated embodiment, the sliding portion 310 has a disc shape. The sliding part 310 is provided in parallel with the separation film 210.

The mounting portion 320 is provided near the first end 111 and can slide toward the second end 113. Further, the mounting portion 320 is partially accommodated in the functional chamber 150 closest to the first end 111 and seals the cylindrical body 100. In the illustrated embodiment, the mounting portion 320 is substantially cylindrical. The mounting portion 320 is located on the side of the sliding portion 310 close to the first side plate 120. One end of the mounting part 320 is fixed to the sliding part 310.

Further, the cylinder 100 is provided with a mounting hole (not shown) communicating with the functional chamber 150 closest to the first end 111. The slider 300 can penetrate the mounting hole and be at least partially received in the functional chamber 150 closest to the first end 111. In the illustrated embodiment, the mounting hole is opened on the first side plate 120. The mounting portion 320 penetrates the mounting hole and is partially accommodated in the functional chamber 150 closest to the first end 111.

The pushing member 400 and the sliding member 300 are detachably connected. Further, the pushing member 400 can slide against the pushing member 300. In the illustrated embodiment, the pushing member 400 has a rod shape. Furthermore, the mounting portion 320 defines a receiving slot 322. The receiving slot 322 is opened at the end of the mounting part 320 away from the sliding part 310. The pushing member 400 can be at least partially received in the receiving groove 322 to slide against the pushing member 300. In the illustrated embodiment, the accommodating groove 322 is formed inward by the end of the mounting portion 320 away from the sliding portion 310.

Further, the separation device 10 further includes an adsorption member (not shown). The suction member is accommodated in the functional chamber 150 closest to the first end 111. The pushing member 400 is at least partially magnetic. When the pushing member 400 is connected to the sliding member 300, the pushing member 400 can provide a magnetic field to the adsorption member, and fix the adsorption member on the sliding member 300.

In the illustrated embodiment, the suction member is accommodated in the functional chamber 150 closest to the first end 111 and located on the side of the sliding part 310 away from the mounting part 320. The side of the pushing member 400 close to the slider 300 has magnetism to provide a magnetic field to the suction member so that the suction member can be fixed on the side of the sliding part 310 away from the mounting part 320. It should be noted that the pushing member 400 may also be directly made of magnetic material.

Optionally, the adsorption member is a magnetic bead. Further, the magnetic beads are superparamagnetic silicon oxide nano magnetic beads or carboxylated magnetic beads. It should be noted that the adsorbent is not limited to the magnetic beads indicated above, but can also be other magnetic beads, which can be set according to the actual situation, as long as it can be combined with the target substance in the sample to be tested.

Optionally, the separation device 10 further includes a lysis solution to lyse the sample to be tested to release the target substance. The lysate is contained in the functional chamber 150 closest to the first end 111. Further, when the target substance is a nucleic acid, the lysate is a cell lysate or a protein lysate. It should be noted that the lysate is not limited to the lysate indicated above, but can also be other lysates, which can be set according to the actual situation, as long as the target substance in the sample to be tested can be released.

The separation device 10 further includes a magnetic field generating mechanism (not shown) configured to provide a magnetic field to the adsorption member. The adsorbent can move under the action of the magnetic field. By adding an external magnetic field, the adsorbent can be moved to achieve the effect of mixing. The magnetic field generating mechanism may be a magnetic field generating mechanism with an electromagnetic coil, or a magnetic field generating mechanism with a permanent magnet.

The waste liquid tank 600 communicates with the liquid discharge hole 160. Further, the waste liquid tank 600 is cylindrical. The outer wall of the waste liquid tank 600 is fixed to the outer wall of the cylinder 100. In the illustrated embodiment, the extending direction of the waste liquid tank 600 is substantially parallel to the extending direction of the cylinder 100. An installation port (not shown) is opened on the outer peripheral surface of the waste liquid tank 600. The outer wall of the waste liquid tank 600 is fixed to the outer wall of the cylinder 100 and shields the installation opening, so that the waste liquid tank 600 and the cylinder 100 cooperate to form a receiving chamber (not shown).

Furthermore, the side wall of the cylinder 100 is further provided with an air pressure balancing hole 144 communicating with the waste liquid tank 600. The air pressure balance hole 144 is disposed near the first end 111. The air pressure balancing hole 144 is configured to balance the air pressure in the cylinder 100. When the sample to be tested needs to be added to the functional chamber 150 closest to the first end 111, the sliding part 310 may be located between the sample port 170 and the air pressure balance hole 144.

The detection device communicates with the separation device 10 to perform qualitative or quantitative detection of the target substance in the sample to be tested. In the illustrated embodiment, the detection device is a PCR (polymerase chain reaction) detection device. Optionally, the detection device is a microfluidic PCR (polymerase chain reaction) detection device. The detection device can be hermetically connected to the sample outlet 190 so that the target substance in the end chamber 140 can enter the detection device for detection.

The operation process of the above separation device 10 is as follows:

(1) During the preparation of the separation device 10, the sliding part 310 is accommodated in the functional chamber 150 closest to the first end 111, and the sliding part 310 is positioned between the sample port 170 and the air pressure balance hole 144, An adsorption member is added to the functional chamber 150 closest to the first end 111, and the adsorption member is located on the side of the sliding part 310 away from the mounting part 320, and a cleaning solution is added to the other functional chamber 150, and the end chamber 140 Add eluent. It should be noted that, if the sample to be tested needs to be cracked to release the target substance to be adsorbed on the adsorption member, the lysis solution is added to the functional chamber 150 closest to the first end 111.

(2) The sample to be tested is added into the functional chamber 150 closest to the first end 111 from the sample loading port 170, and the magnetic field generating mechanism is turned on to provide a magnetic field to the barrel 100, so that the sample to be tested and the adsorbent are mixed evenly. If a lysate is added, the sample to be tested is decomposed under the action of the lysate to release the target substance, and the released target substance is adsorbed with the adsorbent to obtain an adsorbent carrying the target substance.

(3) After the mixing is completed, close the magnetic field generating mechanism. The pushing member 400 is received in the receiving groove 322 to push the sliding member 300 and fix the suction member. The sliding member 300 moves toward the second end 113 by the pushing member 400, and the sealing film 220 and the separating film 210 of the functional chamber 150 where the sliding part 310 is located are sequentially broken under the action of the air pressure, and the waste liquid is discharged from the The liquid hole 160 flows into the waste liquid tank 600, and the sliding member 300 continues to move, so that the adsorption member carrying the target substance enters the next functional chamber 150, and stops moving, so that the pushing member 400 moves toward the first end 111 and cannot Fix the suction element. The magnetic field generating mechanism is turned on to provide a magnetic field to the cylinder 100, so that the adsorbent and the cleaning solution are mixed to clean the adsorbent carrying the target substance.

(4) After cleaning, close the magnetic field generating mechanism. The pushing member 400 is received in the receiving groove 322 to push the sliding member 300 and fix the suction member. The sliding member 300 moves toward the second end 113 by the pushing member 400, and the sealing film 220 and the separating film 210 of the functional chamber 150 where the sliding part 310 is located are sequentially broken under the action of the air pressure, and the waste liquid is discharged from the The liquid hole 160 flows into the waste liquid tank 600, and the sliding member 300 continues to move, so that the adsorption member carrying the target substance enters the next functional chamber 150, and stops moving, so that the pushing member 400 moves toward the first end 111 and cannot Fix the suction piece. The magnetic field generating mechanism is turned on to provide a magnetic field to the cylinder 100, so that the adsorbent and the cleaning solution are mixed to clean the adsorbent carrying the target substance.

(5) After cleaning, close the magnetic field generating mechanism. The pushing member 400 is moved toward the second end 113 and partially received in the receiving groove 322 to push the sliding member 300 and fix the suction member. The sliding member 300 moves toward the second end 113 by the pushing member 400, and the sealing film 220 and the separating film 210 of the functional chamber 150 where the sliding part 310 is located are sequentially broken under the action of the air pressure, and the waste liquid is discharged from the The liquid hole 160 flows into the waste liquid tank 600, and the sliding member 300 continues to move, so that the adsorption member carrying the target substance enters the end chamber 140, and stops moving, so that the pushing member 400 moves toward the first end 111 and cannot be fixed Sorption parts. The magnetic field generating mechanism is turned on to provide a magnetic field to the cylinder 100, so that the adsorbent carrying the target substance is mixed with the eluent to separate the target substance from the adsorbent to obtain a pure target substance.

(6) After the elution, the magnetic field generating mechanism is turned off. The sample outlet 190 is communicated with the detection device. The pushing member 400 is moved toward the second end 113 and partially received in the receiving groove 322 to push the sliding member 300 and fix the suction member. The sliding member 300 moves toward the second end 113 by the pushing member 400. The target substance flows into the detection device under the action of the slider 300 for quantitative and qualitative detection.

The above detection system has at least the following advantages:

In the separation device 10 of the above detection system, the cylinder 100 is divided into an end chamber 140 and N functional chambers 150 by a plurality of separation membranes 210, and the plurality of separation membranes 210 are all sealingly connected to the cylinder 100, N The side walls of the functional chamber 150 are provided with drainage holes 160, and each drainage hole 160 is sealed with a blocking film 220, which can be placed in the functional chamber 150 closest to the first end 111 to be configured as an adsorption target The material adsorbent can place the reaction liquid or cleaning liquid in other functional chambers 150; since the slider 300 is disposed near the first end 111 and can slide toward the second end 113, when the target substance needs to be separated , The slider 300 can be accommodated in the functional chamber 150 closest to the first end, an adsorption member can be provided on the side of the slider 300 away from the first end 111, and the sample to be separated is added to make the target substance adsorb to the adsorption member On the top, slide the slider 300 toward the second end 113 to rupture the blocking membrane 220 and the separation membrane 210, and carry the adsorbent adsorbing the target substance into the other functional chamber 150 for reaction or cleaning, and finally enter End chamber 140 Pure target substance; when the target substance needs to be detected, the pure target substance can be flowed from the end chamber 140 into the detection device for detection, or can be directly detected in the end chamber 140, or at the end The reaction reagent or detection reagent is placed in the chamber 140 so that the target substance reacts with the reaction reagent or detection reagent to thereby detect the target substance. The above-mentioned separation device 10 is simple, and the separation and detection of the target substance can be achieved without repeatedly opening the cover, the operation is convenient, and the sample to be tested is less polluted. The above separation device 10 can be configured for nucleic acid extraction, chemiluminescence detection, or cell sorting.

It can be understood that the separation film 210 is not limited to three, but may be more, for example, four or five.

It can be understood that the waste liquid tank 600 is not limited to the above-mentioned pointed shape, and may also be a ring-shaped hollow structure. When the waste liquid tank 600 has a ring-shaped hollow structure, the cylinder 100 is at least partially accommodated in the waste liquid tank 600 and is sealedly connected to the waste liquid tank 600. It can be understood that the waste liquid tank 600 may also be omitted. When the waste liquid tank 600 is omitted, the waste liquid flowing out of the drain hole 160 can directly flow into the external waste liquid collector.

It can be understood that the detection system is not limited to the PCR (polymerase chain reaction) detection system, but can also be other detection systems, for example: please refer to FIG. 4 together. In other embodiments, the detection system is a chemiluminescence detection system, including separation Device 10 'and chemiluminescence detection device (not shown).

The structure of the separation device 10 'is substantially the same as that of the separation device 10, except that the end chamber 140' is a detection chamber. The functional chamber 150 'closest to the first end 111' is the sample chamber. At least one of the remaining functional chambers 150 'is a reaction chamber.

In the illustrated embodiment, there are four separation films 210 '. Four separation films 210 'are arranged at intervals from the second end 113' to the first end 111 'to divide the cylinder 100' into an end chamber 140 'and four functional chambers 150'. The end chamber 140 'is configured to accommodate the detection reagent. The functional chamber 150 'closest to the first end 111' is the sample chamber. From the first end 111 'to the direction close to the second end 113', the remaining three functional chambers 150 'are arranged in order to contain the cleaning solution, the reaction reagent, and the cleaning solution. The reaction reagent is configured as a reaction product obtained by reacting with the target substance in the sample to be tested, and the reaction product can specifically bind to the detection reagent to emit a signal. Among them, the signal is a light-emitting signal.

The chemiluminescence detection device is connected to the separation device 10 'to detect the target substance in the sample to be tested. In the illustrated embodiment, the chemiluminescence detection device is a light absorption and analysis device. The chemiluminescence detection device is connected to the end chamber 140 'and can receive and analyze the luminescence signal in the end chamber 140' to perform quantitative and qualitative detection of the target substance in the test sample. It should be noted that the light-emitting signal may be a fluorescent signal or other signals.

The operation of the chemiluminescence detection system of the above embodiment is as follows:

(1) During the preparation of the separation device 10 ', the sliding part 310' is accommodated in the functional chamber 150 'closest to the first end 111', and the sliding part 310 'is located at the sample port 170' and the air pressure is balanced Between the holes 144 ', add a suction member in the functional chamber 150' closest to the first end 111 ', and position the suction member on the side of the sliding part 310' away from the mounting part 320 ', from the first end 111' Close to the direction of the second end 113 ', the remaining three functional chambers 150' are added with cleaning solution, incubation reagent and cleaning solution in sequence. The detection reagent is added to the end chamber 140 '. It should be noted that, if the sample to be tested needs to be cracked to release the target substance to be adsorbed on the adsorbent, the lysis solution is added to the functional chamber 150 'closest to the first end 111'.

(2) The sample to be tested is added into the functional chamber 150 'closest to the first end 111' from the sample loading port 170 ', and the magnetic field generating mechanism is turned on to provide a magnetic field to the cylinder 100', so that the sample to be tested is mixed with the adsorbent uniform. If a lysate is added, the sample to be tested is decomposed under the action of the lysate to release the target substance, and the released target substance is adsorbed with the adsorbent to obtain an adsorbent carrying the target substance.

(3) After the mixing is completed, close the magnetic field generating mechanism. The pushing member 400 'is accommodated in the accommodating groove 322' to push the sliding member 300 'and fix the suction member. The sliding member 300 'is moved toward the second end 113' by the pushing member 400 ', and the sealing film 220' and the separation film 210 'of the functional chamber 150' where the sliding part 310 'is located are under the action of air pressure In turn, the waste liquid flows into the waste liquid tank 600 'from the drain hole 170', and the sliding member 300 'continues to move, so that the adsorption member carrying the target substance enters the next functional chamber 150', stops the movement, and causes the pushing member 400 'Move toward the first end 111' without fixing the suction member. The magnetic field generating mechanism is turned on to provide a magnetic field to the cylinder 100 ', so that the adsorption member and the cleaning solution are mixed to clean the adsorption member carrying the target substance.

(4) After cleaning, close the magnetic field generating mechanism. The pushing member 400 'is accommodated in the accommodating groove 322 to push the sliding member 300' and fix the suction member. The sliding member 300 'is moved toward the second end 113' by the pushing member 400 ', and the sealing film 220' and the separation film 210 'of the functional chamber 150' where the sliding part 310 'is located are under the action of air pressure In turn, the waste liquid flows into the waste liquid tank 600 'from the drain hole 170', and the sliding member 300 'continues to move, so that the adsorption member carrying the target substance enters the next functional chamber 150', stops the movement, and causes the pushing member 400 'Move toward the first end 111' without fixing the suction member. The magnetic field generating mechanism is turned on to provide a magnetic field to the cylinder 100 ', so that the adsorption member and the reaction reagent are uniformly mixed to obtain the adsorption member carrying the reaction product.

(5) After the reaction is over, close the magnetic field generating mechanism. The pushing member 400 'is accommodated in the accommodating groove 322 to push the slider 300' and fix the suction member. The sliding member 300 'is moved toward the second end 113' by the pushing member 400 ', and the sealing film 220' and the separation film 210 'of the functional chamber 150' where the sliding part 310 'is located are under the action of air pressure In turn, the waste liquid flows into the waste liquid tank 600 'from the drain hole 170', and the sliding member 300 'continues to move, so that the adsorption member carrying the target substance enters the next functional chamber 150', stops moving, and causes the pushing member 400 'Move toward the first end 111' without fixing the suction member. The magnetic field generating mechanism is turned on to provide a magnetic field to the cylinder 100 ', so that the adsorption member and the cleaning solution are mixed to clean the adsorption member carrying the reaction product.

(6) After cleaning, close the magnetic field generating mechanism. The pushing member 400 'is moved toward the second end 113' and partially accommodated in the receiving groove 322 'to push the sliding member 300' and fix the suction member. The sliding member 300 'is moved toward the second end 113' by the pushing member 400 ', and the sealing film 220' and the separation film 210 'of the functional chamber 150' where the sliding part 310 'is located are under the action of air pressure In turn, the waste liquid flows into the waste liquid tank 600 'from the drain hole 170', and the sliding member 300 'continues to move, so that the adsorption member carrying the reaction product enters the end chamber 140', stops the movement, and causes the pushing member 400 ' Move toward the first end 111 'without fixing the suction member. Turn on the chemiluminescence detection device. The magnetic field generating mechanism is turned on to provide a magnetic field to the cylinder 100 ', so that the adsorption member carrying the reaction product and the detection reagent are mixed and reacted. At the same time, the chemiluminescence detection device collects and analyzes the luminescence signal during the reaction between the adsorbent carrying the reaction product and the detection reagent to perform quantitative and qualitative analysis on the target substance in the sample to be tested.

The chemiluminescence detection system of the above embodiment has at least the following advantages:

The separation device 10 'in the above chemiluminescence detection system has a simple structure, and the target substance can be separated without repeatedly opening the lid. The operation is convenient, and the sample to be tested is less polluted. At the same time, the above chemiluminescence detection system is used to detect the The process can be directly detected without removing the target substance from the separation device 10 ', which further reduces the contamination of the target substance and ensures the accuracy of the detection.

It can be understood that if the target substance in the sample to be tested can directly specifically bind to the detection reagent, the reaction reagent may be omitted. It can be understood that the separation film 210 'is not limited to four, but may be more, such as five or six, etc., and may be set according to actual conditions.

It can be understood that the sample outlet 190 'may be omitted.

It can be understood that the detection system is not limited to PCR (polymerase chain reaction) detection system and chemiluminescence detection system, but can also be other detection systems, for example: in other embodiments, the detection system is an electrochemical detection system (not shown) , Including separation device and electrochemical detection device. The structure of the separation device and the separation device 10 are substantially the same, the difference is that:

The adsorbent is a chip. The adsorption member is accommodated in the functional chamber closest to the first end and fixed on the sliding member. The pusher can be made of non-magnetic material. The magnetic field generating mechanism is omitted. The electrochemical detection device can collect and analyze the electrical signal of the end chamber to perform quantitative and qualitative analysis on the target substance in the sample to be tested.

The separation device in the above-mentioned electrochemical detection system has a simple structure, and the target substance can be separated and detected without repeatedly opening the cover. The operation is convenient, and the sample to be tested is less polluted to ensure the purity of the target substance.

It can be understood that the above separation device 10 is not limited to being configured as a detection system or an electrochemical detection system, but can also be configured into other separation systems, for example: a cell sorting system (not shown) of an embodiment, including a separation device and cells Detection device. The separation device of the cell sorting system is almost the same as the separation device 10, except for:

The cleaning solution in the functional chamber is a cleaning solution or a separation solution capable of cleaning the target cells and ensuring the activity of the target cells. The end chamber is configured with a buffer solution to suspend the target cells.

Among them, the adsorption member is a biological magnetic bead. The magnetic beads can specifically bind to the recognition sites on the surface of the cells to be separated. Specifically, the magnetic beads are MACS MicroBeads magnetic beads or Dynabeads magnetic beads. The target cells in the cells to be separated are hybridoma cells or CAR-T cells (chimeric antigen receptor T cell immunotherapy, English full name Chimeric Antigen Receptor T-Cell Immunotherapy). It should be noted that the adsorbent may also be a chip that can specifically bind to the target cell. When the adsorption member is a chip, the adsorption member is accommodated in the functional chamber closest to the first end and fixed on the sliding member.

The cell detection device is connected to the separation device to perform quantitative or qualitative separation of target cells in the cells to be separated. Further, the cell detection device is connected to the end chamber to perform qualitative or quantitative detection on target cells in the end chamber. Furthermore, the cell detection device can detect and analyze the concentration, purity, and type of target cells.

Among them, the cell detection device is a spectrophotometer, a Raman spectrometer or a chemiluminescence detector. It should be noted that the cell detection device is not limited to the above-mentioned pointing device, as long as the device capable of quantitatively and qualitatively detecting the target cell can be used as the cell detection device, and can be set according to the actual situation.

With the simple structure of the separation device in the cell sorting system described above, the separation and detection of target cells can be achieved without repeated opening of the cover, the operation is convenient, and the sample to be tested is less polluted to ensure the purity of the target cells.

The technical features of the above-mentioned embodiments can be arbitrarily combined. To simplify the description, all possible combinations of the technical features in the above-mentioned embodiments are not described. However, as long as there is no contradiction in the combination of these technical features, All should be considered within the scope of this description.

The above-mentioned embodiment only expresses several implementation manners of the present application, and its description is more specific and detailed, but it should not be understood as limiting the scope of the patent application. It should be noted that, for those of ordinary skill in the art, without departing from the concept of the present application, a number of modifications and improvements can also be made, which all fall within the protection scope of the present application. Therefore, the protection scope of the patent of this application shall be subject to the appended claims.

Industrial applicability

The separation device and its application, detection system, electrochemical detection system, and cell sorting system provided in the embodiments of the present application are simple in structure through the separation device in the above cell sorting system, and the target sample can be separated without repeatedly opening the cover And detection, easy operation, less contamination of the sample to be tested.

Claims

A separating device, characterized in that it includes: a cylinder body having opposite first and second ends;

The diaphragm assembly includes a plurality of separation membranes and a plurality of sealing membranes. The plurality of separation membranes are all accommodated in the cylinder body and are all sealedly connected to the cylinder body. The plurality of separation membranes extend from the second end Spaced toward the first end to divide the barrel into an end chamber and N functional chambers, the end chamber is close to the second end, the sidewalls of the N functional chambers There are drain holes on the openings, and each of the drain holes is blocked with the blocking film; and

The slider is provided near the first end and can slide toward the second end.
The separating device according to claim 1, wherein the separating device further comprises a pushing member, and the pushing member is detachably connected with the sliding member.
The separation device according to claim 2, further comprising an adsorption member, the adsorption member being accommodated in the functional chamber closest to the first end, the pushing member is at least partially magnetic, so When the pushing member is connected to the sliding member, the pushing member can adsorb the adsorption member, and the adsorption member can be adsorbed on the sliding member.
The separation device according to claim 3, wherein the separation device further includes a magnetic field generating mechanism configured to provide a magnetic field to the adsorption member, the adsorption member being movable under the action of the magnetic field.
The separation device according to claim 3 or 4, wherein the adsorption member is a magnetic bead.
The separation device according to claim 5, wherein the magnetic beads are superparamagnetic silicon oxide nano-magnetic beads or carboxylated magnetic beads.
The separation device according to any one of claims 1-6, further comprising an adsorption member, wherein the adsorption member is a chip, and the adsorption member is accommodated in the functional chamber closest to the first end And fixed to the slider.
The separation device according to any one of claims 1-7, wherein the tensile strength of the separation membrane is greater than the tensile strength of the blocking membrane, and the sliding member is closer to the second end When sliding in the direction, the pressure in the functional chamber where the slider is located can be increased, and the blocking film and the separation membrane in the functional chamber where the slider is located can be sequentially ruptured.
The separation device according to any one of claims 1-8, further comprising a waste liquid tank, the waste liquid tank communicating with the liquid discharge hole.
The separation device according to claim 9, characterized in that the waste liquid tank is a ring-shaped hollow structure, and the cylinder body is at least partially accommodated in the waste liquid tank, and is sealingly connected to the waste liquid tank;

Or, the waste liquid tank is cylindrical, and the outer wall of the waste liquid tank is fixed to the outer wall of the barrel body.
The separation device according to claim 9 or 10, wherein a side wall of the cylinder is further provided with an air pressure balancing hole communicating with the waste liquid tank, and the air pressure balancing hole is provided near the first end .
The separation device according to any one of claims 1-11, wherein the cylinder body is provided with a sampling port, and the sampling port is located in the cavity of the functional chamber closest to the first end On the wall, the separation device further includes a seal configured to seal the sample port;

The cylinder body is provided with a sample outlet, the sample outlet is communicated with the end chamber, and the separation device further includes a stopper configured to seal the sample outlet.
The separation device according to any one of claims 1-12, wherein the barrel includes a barrel body, a first side plate, and a second side plate;

The first end and the second end are provided with a first opening and a second opening, the first opening is opposite to the second opening, and the first side plate and the second side plate respectively shield the The first opening and the second opening are both hermetically connected to the barrel body.
The separation device according to claims 1-13, characterized in that the sliding member comprises a sliding part and a mounting part fixed to the sliding part;

The sliding portion is provided near the first end and can slide toward the second end; the sliding portion is housed in the functional chamber near the first end and is connected to the body of the cylinder Sealed connection on the inner wall;

The mounting portion is provided near the first end and can slide toward the second end; the mounting portion is partially accommodated in the functional chamber near the first end and can seal the cylinder.
The application of the separation device according to any one of claims 1 to 14 in nucleic acid extraction, chemiluminescence detection or cell sorting.
A detection system, characterized by comprising the separation device according to any one of claims 1-6 and 8-14.
The detection system according to claim 16, wherein the detection system is a PCR detection system, the end chamber is a detection chamber, and the functional chamber closest to the first end is a sample chamber, At least one of the remaining functional chambers is a cleaning chamber;

Or, the detection system is a chemiluminescence detection system, the end chamber is a detection chamber, the functional chamber closest to the first end is a sample chamber, and at least one of the remaining functional chambers For the reaction chamber.
An electrochemical detection system, characterized by comprising the separation device according to any one of claims 1-2 and 7-14.
The electrochemical detection system according to claim 18, wherein the end chamber is a detection chamber, the functional chamber closest to the first end is a sample chamber, and the remaining functional chambers At least one of them is a cleaning chamber.
A cell sorting system, characterized by comprising the separation device according to any one of claims 1-14.