WO2022184135A1

WO2022184135A1 - Automatic device for effective enrichment of neurogenic exosomes in blood

Info

Publication number: WO2022184135A1
Application number: PCT/CN2022/079045
Authority: WO
Inventors: 马咏翔; 李佳
Original assignee: 北京尔瑞鑫悦科技有限公司
Priority date: 2021-03-05
Filing date: 2022-03-03
Publication date: 2022-09-09
Also published as: CN113025480B; CN113025480A

Abstract

An automatic device for effective enrichment of neurogenic exosomes in blood, comprising: a reaction rack at the bottom and a movable operation frame above the reaction rack; and a control structure for controlling the operation frame to operate according to a predetermined track and controlling the reaction rack to react according to a set program. The reaction rack is provided with a shaking incubation structure required for implementing enrichment of exosomes, an exocentric structure, a reagent placement structure, a consumable material placement structure, a discard placement structure, a sample placement structure, sample information identification structure, and a reacted enrichment sample collection structure. The operation frame is provided with a tube moving structure for implementing the moving of a tube between different structures and a liquid taking and adding structure for implementing liquid transfer in a whole process. In addition, the reaction rack or the operation frame is provided with an EP tube marking and identification structure for a first use of an EP tube; the operation frame is controlled by the control structure to move and clamp a sample tube and different liquids, such that automatic enrichment of samples is implemented according to the set program.

Description

An automated device for efficient enrichment of neuroderived exosomes in blood

technical field

The invention belongs to the technical field of medical devices, in particular to an automatic enrichment device for exosome enrichment; in particular, it relates to an automatic device for effectively enriching neurogenic exosomes in blood.

Background technique

Neurodegenerative diseases are caused by the loss of neurons and/or their myelin sheaths, which worsen and become dysfunctional over time. Such as Alzheimer's disease (AD), Parkinson's disease (PD), and prion disease. At present, the definite clinical diagnosis of neurodegenerative diseases relies on PET (positron emission tomography), and the disease often develops to an irreversible stage when pathological changes occur.

The detection of relevant biomarkers in cerebrospinal fluid can advance the diagnosis of such diseases, but the sampling method of cerebrospinal fluid is very traumatic to the human body, and the patient's willingness is not high; very few patients will be tested in advance before symptoms appear , This kind of test is often performed after the patient has symptoms, and the collection of cerebrospinal fluid is difficult, high-risk, and has high requirements for operators.

Therefore, it is necessary to find a method that is simple to collect and easy to detect, preferably one screening method in ordinary annual physical examination. With the in-depth study of exosomes, it is found that neurogenic exosomes in blood can be used as samples to detect relevant indicators of neurodegenerative diseases, so that the diagnosis of such diseases can not only be greatly advanced, but also compared with cerebrospinal fluid. , such samples are more compliant with subjects.

However, neurogenic exosomes in the blood need to be enriched to obtain them. Although the current technology can obtain such samples, it is completed by manual operation. The entire operation process includes multiple processes such as cooling, centrifugation, vibration, sample addition, and blowing, which takes a long time and requires the operator to perform timing. , the time is long, and the purely manual operation is prone to wrong conclusions due to sample errors in the operation process, so it cannot be an effective way to quickly generalize and obtain conclusions effectively.

If there is an automated device to complete the above process, quickly and effectively enrich the subject's neurogenic exosomes, and detect relevant information, it will be known in advance whether the subject is at risk of developing such diseases. and prevent it in advance. The promotion and application of this detection method will greatly reduce the occurrence of excessive neurodegenerative diseases along with the aging of the population.

Aiming at the problem in the prior art that there is no effective and rapid enrichment equipment for blood neurogenic exosomes, the present invention provides an automated device for effectively enriching neurogenic exosomes in blood.

SUMMARY OF THE INVENTION

An automated device for effectively enriching neurogenic exosomes in blood, comprising a reaction rack at the bottom and a movable operating rack above; and a control for controlling the operating rack to run according to a predetermined track and the reaction rack to respond according to a set program It is characterized in that the reaction rack is provided with a shaking incubation structure, a centrifugal structure, a reagent placement structure, a consumables placement structure, a discard placement structure, a sample placement structure, a sample information identification structure, a reaction After the enrichment sample collection structure; the operation rack is provided with a tube body moving structure that realizes the movement of the tube body between different structures, and realizes the liquid taking and adding liquid structure for liquid transfer in the whole process; in addition, on the reaction rack or the operation rack An EP tube marking and identification structure is provided when the EP tube is used for the first time; the device also includes a storage module capable of storing information, and the control structure controls the oscillating speed and amplitude of the oscillating incubation structure, and controls the temperature required for the corresponding steps of the oscillating incubation structure; The control structure controls and monitors the amount of reagents in the reagent placement structure, and prompts for replacement when there is no reagent; the control structure also controls the centrifugal parameters of the centrifugal structure; the control structure monitors and records the information of the samples in the placement holes corresponding to the sample placement structure; the control structure controls the sample The information identifies the sample tube information and stores it in the storage module; the control structure controls the movement of the liquid taking and adding tube to perform the process of taking liquid and adding liquid, and the control structure controls the movement of the tube body. The structure grabs the corresponding tube body and moves to the corresponding next structure. In the area, the control structure controls the EP tube marking and identification structure to scan or identify the information on the EP tube and correspond to the corresponding sample tube information; avoid confusion or loss of patient information during the movement process; the control structure controls the last EP tube to be set to The enrichment sample was collected in the structure.

During the specific operation, the control structure control information identification structure first identifies the information code of the sample tube and stores it in the storage unit of the equipment, and then sets it in the sample placement structure in sequence after identification; then moves the structure through the tube body on the operation rack. Grab the sample tubes in the sample placement structure according to the set order and reach the next processing structure in the oscillation incubation structure. After entering the oscillation incubation structure, the liquid taking and adding structure moves to the consumables placement structure according to the control settings, and the corresponding consumables are installed. Then move to the reagent placement structure. After obtaining the corresponding reagents, move to the corresponding sample tube placement place of the shaking incubation structure for reagent addition. After the reagent is added, the liquid-taking and liquid-adding structure leaves and moves to the discard placement structure. Discard the waste consumables in The discards are placed in the structure; after adding the reagent, the control structure controls the shaking incubation structure to perform shaking or incubation processing. The steps that need to add reagents during the shaking incubation process are all reagent additions according to the above steps; when the sample tube needs to be moved into the centrifuge structure , the control structure controls the movement of the tube body to grab the sample tube to move and place it in the centrifugal structure for centrifugation. If liquid needs to be added after centrifugation, add reagents according to the above steps of taking liquid and adding liquid. When moving to the EP tube, first place the EP tube in the structure of the next step, scan or identify the information on the EP tube through the EP tube marking and identification structure, and correspond to the corresponding sample tube information, and then use the liquid sampling and adding structure. Move the liquid in the sample tube to the corresponding EP tube. When the EP tube is in the shaking incubation structure, the shaking incubation is carried out according to the shaking incubation process. When the reagent needs to be added, add the reagent. When it needs to be centrifuged again, use the tube body moving structure to remove it. Move it to the centrifuge structure; when the sediment needs to be retained after the centrifugation, the control structure controls the liquid extraction structure to extract the supernatant from the EP tube, and discard the supernatant in the discard placement structure; follow the steps of exosome enrichment After the enrichment is completed under the control of the control structure, the enriched EP tube is closed and placed into the enrichment sample collection structure.

Further, according to the operation steps, multiple oscillating incubation structures, centrifugation structures, reagent placement structures, consumables placement structures, and discard placement structures need to be set up; in this way, the upper operation rack can reduce the moving distance and increase the continuity of the entire automation process.

Further, if the sample or EP tube needs to add reagents during the shaking incubation structure stage, set up the corresponding reagent placement structure, consumables placement structure, and discard placement structure next to the corresponding shaking incubation structure, so that it is convenient to obtain materials nearby and discard them nearby.

Or, if only one batch of samples is enriched in one operation, when the structure and shape of the sample tube and the EP tube are the same, one oscillation incubation structure and one centrifugal structure are set up in the whole process; With little space, this kind of equipment is mainly aimed at the enrichment process of relatively small samples and can be used in the laboratory.

Or, if only one batch of sample enrichment is performed in one operation, when the shape of the sample tube and the EP tube are different, the minimum number of shaking incubation structures and centrifugation structures should be set up according to the requirements of the tubes of different shapes. Enrichment of batch samples.

Further, for the case where only one batch of samples is enriched in a process, set up a reagent placement structure, consumables placement structure, and discard placement structure, and set all reagent placement structures, consumables placement structures, and discards for exosome enrichment in a unified manner. The object placement structure can further reduce the space occupied by the equipment, and because the overall structure is small, the movement of the operation rack does not take too much time.

Further, the operating rack is arranged on the top of the setting, which is a setting top whose position relative to the reaction rack above the reaction rack does not change, and a moving track is provided on the setting top to ensure that the operating rack can move above any structure and perform corresponding operations.

Further, the operation frame is an integral frame, on which a liquid-taking and liquid-adding structure and a tube body moving structure are arranged, and the liquid-taking and liquid-adding structure and the tube body moving structure can move up and down on the operation frame, so that one structure can play its own role. structure.

Alternatively, two operation racks are provided, one for taking liquid and adding liquid and the other for moving the pipe body; when they are functioning, they are moved to the top of the corresponding structure for operation. When one operation rack is in action, the other operation rack passes through The moving rail moves to a position where it will not interfere with another operating frame.

Further, the same number of operation heads are set on the liquid taking and liquid adding structure and the tube body moving structure, so that the same number of sample tubes can be clamped and the reagents can be added correspondingly.

Further, the reagent placement structure corresponds to a single reagent, and the number of liquid intake ports equal to the number of operation heads of the liquid intake and liquid addition structure is set, and the positional relationship of the liquid intake ports is consistent with the positional relationship of the operation head; and the consumables placement structure is set for the corresponding single consumable. The number of sampling ports is equal to the number of operation heads of the liquid sampling and liquid adding structure; and the positional relationship of the sampling ports is consistent with the positional relationship of the operation heads.

Further, the number of placement holes in a single column is an integer multiple of the number of operation heads, or the number of columns of placement holes is the same as the number of operation heads, so as to ensure that the sample tubes are obtained in an effective order.

Further, a fault alarm structure is also set between each structure and the control structure, when the corresponding fault alarm structure sends out an alarm signal, the corresponding structure is prompted to send a fault. Or, the control structure is also connected to a display structure, the display structure displays the corresponding alarm structure alarm, and prompts the corresponding structure failure to be repaired.

Further, the number of reactions corresponding to the reagent amounts of any two different reagents in the reagent placement structure is equal, and when the number of times is reached, a prompt or display on the display structure is displayed for replacement.

Further, the reaction times corresponding to the quantities of any two different consumables in the consumable placement structure are also equal, or the quantity of each consumable is the same as the number of reactions of the reagents, which facilitates the overall replacement of reagents and consumables.

Further, the discarded object placement structure is divided into a consumables discarded part and a liquid discarded part; the discarded consumables and the liquid are treated respectively.

Further, the amount of discarded objects accommodated by the discarded object placement structure is equal to the space required to accommodate one replacement of reagent consumables, which can be replaced together with the reagent consumables, which can ultimately reduce monitoring difficulty and increase efficiency.

Further, the reagent placement structure, the consumables placement structure and the discard placement structure are arranged under the setting plate in the middle of the reaction rack, and are combined under the setting plate for easy replacement.

Further, the EP tube marking and identification structure are arranged on the moving structure of the tube body, and the top of the EP tube is provided with different identification structures that can correspond one by one; by scanning and identifying the EP tube above, and corresponding to the corresponding sample tube information, To achieve the purpose of information matching, the process is simple and convenient.

Further, because the entire enrichment process includes multiple shaking incubation and centrifugation processes, each process includes a shaking incubation and centrifugation step; the reaction rack of the entire equipment is divided into multiple corresponding reaction areas according to the number of shaking incubation and centrifugation processes; Further, each corresponding area is provided with an oscillation incubation structure, a centrifugation structure, a reagent placement structure, a consumables placement structure, and a discard placement structure.

Further, a set of operation racks with a liquid taking and adding liquid structure and a tube body moving structure are arranged above each reaction area. In this way, a streamlined liquid extraction and clamping of the tube body can be realized, and there will be no problem of conflict of use.

Further, the moving track of the operation rack is a track that can satisfy the movement of the operation rack to achieve liquid extraction and gripping of the pipe body.

Further, the moving track is also arranged in different areas, so as to realize the moving track of moving in this area and obtaining the sample or EP tube in the moving area.

Further, the EP tube consumables placement structure is set in the enrichment sample collection structure, or the EP tube consumables placement structure is set to the nearest other reaction area; when the last step is performed, the EP tube is moved to the enrichment sample through the tube body moving structure The domains were collected, and then the enriched supernatant was transferred to an EP tube through a liquid extraction and addition structure for collection, followed by detection.

Application of an automated device for effectively enriching neurogenic exosomes in blood in the enrichment of exosomes in blood.

Compared with the devices of the prior art, the technical solution of the present invention can well solve the problem of enrichment of exosomes in the prior art, and can be completed automatically, so that the detection of exosomes can be well promoted. The promotion of appropriateness can be used to detect neurodegenerative exosomes in blood in advance, diagnose neurodegenerative diseases in advance, and intervene in the effect of treatment, and solve the problem of neurodegenerative diseases in the Chinese population with the occurrence of aging population. The proportion of disease patients will reduce the pressure on the whole society.

Description of drawings

Fig. 1 is the overall structure schematic diagram of the present invention;

Fig. 2 is the top view structure schematic diagram of reaction rack of the present invention;

Fig. 3 is the structural schematic diagram of the side view of part of the operation frame of the present invention;

4 is a schematic diagram of the overall structure of the present invention including only one reaction zone;

FIG. 5 is a schematic structural diagram of the work flow of the equipment of the present invention;

6 is a schematic diagram of the modular structure of the device structure of the present invention;

In the figure, 1. Reaction rack; 11. Vibration incubation structure; 12. Centrifugation structure; 13. Reagent placement structure; 14. Consumables placement structure; 15. Discard placement structure; 16. Sample placement structure; 17. Sample information identification structure; 18. Enriched sample collection structure; 2. Operation rack; 21. Tube movement structure; 24. Setting top; 25. Operating head; 26. Moving track; 3. Control structure; 4. Storage module; 5. Fault alarm structure.

Detailed ways

Example 1

An automated device for effectively enriching neurogenic exosomes in blood, refer to Figures 3-4, 6; it includes a reaction rack 1 at the bottom and a movable operation rack 2 above; and controls the operation rack 2 according to a predetermined track. Operation and control structure 3 of reaction rack 1 according to the set program; reaction rack 1 is provided with shaking incubation structure 11, centrifugation structure 12, reagent placement structure 13, consumables placement structure 14 required to achieve exosome enrichment, and discarded The object placement structure 15, the sample placement structure 16, the sample information identification structure 17, and the enriched sample collection structure 18 after the reaction; the operation rack 2 is provided with a tube body moving structure 21 that realizes the movement of the tube body between different structures. The liquid taking and adding structure 22 for liquid transfer in the whole process; in addition, the EP tube marking and identification structure 23 for the initial use of the EP tube is provided on the reaction rack 1 or the operation rack 2; the device also includes a storage module capable of storing information 4. The control structure 3 controls the oscillating rotation speed and amplitude of the oscillating incubation structure 11, and controls the temperature required for the corresponding steps of the oscillating incubation structure 11; the control structure 3 controls the amount of reagents in the monitoring reagent placement structure 13, and prompts for replacement when there is no reagent; control The structure 3 also controls the centrifugal parameters of the centrifugal structure 12; the control structure 3 monitors and records the information of the samples in the corresponding placement holes of the sample placement structure 16; the control structure 3 controls the sample information to identify the sample tube information and store it in the storage module 4; the control structure 3. Control the movement of the liquid-taking and adding pipe to carry out the liquid-taking and liquid-adding process. The control structure 3 controls the pipe body moving structure 21 to grab the corresponding pipe body and move it to the corresponding area of the corresponding next structure. The control structure 3 controls the EP tube marking and The identification structure 23 scans or identifies the information on the EP tube and corresponds to the corresponding sample tube information; ensures that the patient information is confused or lost during the moving process; the control structure 3 controls the final EP tube to be set in the enrichment sample collection structure 18 .

During the specific operation, the control structure 3 controls the information identification structure to first identify the information code of the sample tube, store it in the storage unit of the device, and set it in the sample placement structure 16 in sequence after identification; The body moving structure 21 grabs the sample tubes in the sample placement structure 16 according to the set order and reaches the next processing structure in the shaking incubation structure 11. After entering the shaking incubation structure 11, the liquid taking and adding structure 22 moves to the consumables according to the control settings. Placement structure 14, install the corresponding consumables, and then move to the reagent placement structure 13. After obtaining the corresponding reagent, move to the corresponding sample tube placement position of the shaking incubation structure 11 for reagent addition. After the reagent is added, the liquid-taking and liquid-adding structure 22 moves away. Go to the discard placement structure 15, and discard the waste consumables in the discard placement structure 15; after adding the reagent, the control structure 3 controls the shaking incubation structure 11 to perform shaking or incubation processing. The steps of adding reagents during the shaking incubation processing are as follows. Reagents are added in the above steps; when the sample tube needs to be moved into the centrifugal structure 12, the control structure 3 controls the tube body moving structure 21 to grab the sample tube to move and place it in the centrifugal structure 12 for centrifugation. Add reagents in the above-mentioned steps of taking liquid and adding liquid. When it is necessary to retain the supernatant and move the supernatant to the EP tube, first place the EP tube in the structure of the next step, and scan or identify the EP through the EP tube marking and identification structure 23. The information on the tube corresponds to the information of the corresponding sample tube, and then the liquid in the sample tube is moved to the corresponding EP tube by using the liquid taking and adding structure 22. When the EP tube is in the shaking incubation structure 11, follow the shaking incubation process. Incubate with shaking, add reagents when needed, and use the tube moving structure 21 to move it into the centrifugal structure 12 when it needs to be centrifuged again; when the sediment needs to be retained after the centrifugation, the control structure 3 controls the liquid taking and adding liquid structure 22 Extract the supernatant from the EP tube, and discard the supernatant in the discard placement structure 15; after the enrichment is completed under the control of the control structure 3 according to the steps of exosome enrichment, the enriched EP tube is closed and placed in the The enrichment sample is collected in structure 18.

If the sample or EP tube needs to add reagents during the 11 stage of the shaking incubation structure, the corresponding reagent placing structure 13, consumables placing structure 14, and discarding structure 15 are set next to the shaking incubation structure 11, so that it is convenient to collect materials and discard them nearby. .

In the case where only one batch of samples is enriched in one operation, when the structure and shape of the sample tube and the EP tube are the same, one shaking incubation structure 11 and one centrifugal structure 12 are set up in the whole process; With little space, this kind of equipment is mainly aimed at the enrichment process of relatively small samples and can be used in the laboratory.

For the case where only one batch of samples is enriched in one process, a reagent placement structure 13, consumables placement structure 14, and discard placement structure 15 are set up, and all reagents, consumables, and discard structures for exosome enrichment are set uniformly, respectively. The space occupied by the equipment is further reduced, and because the overall structure is small, the movement of the operation frame 2 does not take too much time.

The embodiment of the operation rack is as follows: the operation rack 2 is arranged on the setting top 24, the setting top 24 is the setting top 24 whose position relative to the reaction rack 1 above the reaction rack 1 does not change, and the setting top 24 is provided to ensure that the operation rack 2 can move. Move track 26 to go over any structure and perform corresponding operations.

The operation frame 2 is an integral frame, on which a liquid-taking and liquid-adding structure 22 and a tube body moving structure 21 are arranged, and the liquid-taking and liquid-adding structure 22 and the tube body moving structure 21 can move up and down on the operation frame 2, so that a structure A structure that plays its part.

The liquid taking and adding structure 22 and the tube body moving structure 21 are provided with the same number of operation heads 25, so that the same number of sample tubes can be clamped and the reagents can be added accordingly.

The reagent placement structure 13 corresponds to a single reagent and sets the liquid intake ports equal to the number of the operation heads 25 of the liquid intake and liquid addition structure 22, and the positional relationship of the liquid intake ports is consistent with the position relationship of the operation head 25; and the consumables placement structure 14 corresponds to the corresponding single Each consumable is provided with material sampling ports equal to the number of the operation heads 25 of the liquid sampling and liquid addition structure 22 ;

The number of placement holes in a single row is an integer multiple of the number of operation heads 25 , or the number of rows of placement holes is the same as the number of operation heads 25 , to ensure that sample tubes are obtained in an effective order.

The reagent placement structure 13 , the consumables placement structure 14 and the discard placement structure 15 are arranged under the middle setting plate of the reaction rack 1 , and are combined under the setting plate for easy replacement.

The EP tube marking and identification structure 23 is arranged on the tube body moving structure 21, and the top of the EP tube is provided with different identification structures that can correspond one-to-one; To achieve the purpose of information matching, the process is simple and convenient.

The enrichment sample collection structure 18 is provided with the EP tube consumables placement structure 14; in the final step, the EP tube is moved to the enrichment sample collection structure 18 domain by the tube body moving structure 21, and then the enriched supernatant is passed through The liquid taking and adding structure 22 is moved to the EP tube for collection, and then the detection is carried out.

Example 2

On the basis of Example 1, the following embodiments are adjusted:

Referring to FIG. 4 , in the case where only one batch of sample enrichment is performed at a time, an embodiment is added when the shape of the sample tube and the EP tube are different, and the minimum number of shaking incubation structures 11 and centrifugal structures 12 are set according to the requirements of different shapes of tubes. , only one batch of samples is enriched in one process of the whole device. For the case where only one batch of samples is enriched in one process, a reagent placement structure 13, consumables placement structure 14, and discard placement structure 15 are set up, and all reagents, consumables and discard structures for exosome enrichment are set up in a unified manner. The space occupied by the equipment is further reduced, and because the overall structure is small, the movement of the operation frame 2 does not take too much time.

In the embodiment of adding two operation racks, two operation racks 2 are provided, which are respectively a liquid taking and liquid adding operation rack 2 and a pipe body moving operation rack 2; when they play a role, they are respectively moved to the top of the corresponding structure for operation. During the operation of the operation frame 2 , the other operation frame 2 moves to a position where it will not interfere with the other operation frame 2 through the moving rail 26 .

In order to ensure the safety of use, the following embodiments are added: a fault alarm structure 5 is also set between each structure and the control structure 3, and after the corresponding fault alarm structure 5 sends out an alarm signal, the corresponding structure is prompted to send a fault. Or, the control structure 3 is also connected to a display structure, the display structure displays the corresponding alarm structure alarm, and prompts the corresponding structure failure to be repaired.

The number of times of the reagents in the reagent placement structure is limited, specifically: the reaction times corresponding to the reagent amounts of any two different reagents in the reagent placement structure 13 are equal. The reaction times corresponding to the consumables of any two different consumables in the consumables placement structure 14 are also equal, or the consumables of each consumable are the same as the reaction times of the reagents, which facilitates the overall replacement of reagents and consumables. The discarded object placement structure 15 is divided into a consumables discarding part 151 and a liquid discarding part 152; the discarded consumables and the liquid are processed respectively. The amount of discarded objects contained in the discarded object placement structure 15 is equal to the space required to accommodate one replacement of the reagent consumables, which can be replaced together with the reagent consumables, which can ultimately reduce the difficulty of monitoring and increase the efficiency.

Example 3

On the basis of Example 1-2, the following embodiments were adjusted. Refer to Figure 1-2;

Because the entire enrichment process includes multiple shaking incubation and centrifugation processes, each process includes a shaking incubation and centrifugation step; the embodiment of setting multiple reaction areas is specifically: according to the number of shaking incubation and centrifugation processes, the reaction of the entire device is The rack 1 is divided into a plurality of corresponding reaction areas; each corresponding area is provided with an oscillation incubation structure 11 , a centrifugal structure 12 , a reagent placement structure 13 , a consumables placement structure 14 , and a discard placement structure 15 . A set of operation racks 2 with a liquid taking and adding structure 22 and a tube body moving structure 21 is arranged above each reaction area. In this way, a streamlined liquid collection and clamping of the tube body can be realized, and there will be no problem of use conflict. The moving track 26 of the operation frame 2 is a track that can satisfy the movement of the operation frame 2 to realize liquid extraction and clamping of the tube body. The moving rails 26 are also arranged in different areas, which are the moving rails 26 that can realize the movement in this area and the acquisition of samples or EP tubes in the upward moving area.

For the case where only one batch of samples is enriched in one process, a reagent placement structure 13, consumables placement structure 14, and discard placement structure 15 are set up, and all reagents, consumables and discard structures for exosome enrichment are set up in a unified manner. The space occupied by the equipment is further reduced, and because the overall structure is small, the movement of the operation frame 2 does not take too much time.

A fault alarm structure 5 is also set between each structure and the control structure 3. When the corresponding fault alarm structure 5 sends out an alarm signal, the corresponding structure is prompted to send a fault. Or, the control structure 3 is also connected to a display structure, the display structure displays the corresponding alarm structure alarm, and prompts the corresponding structure failure to be repaired.

The reaction times corresponding to the reagent amounts of any two different reagents in the reagent placement structure 13 are equal, and when the times are reached, a prompt is displayed or a replacement is displayed on the display structure. The reaction times corresponding to the consumables of any two different consumables in the consumables placement structure 14 are also equal, or the consumables of each consumable are the same as the reaction times of the reagents, which facilitates the overall replacement of reagents and consumables. The discarded object placement structure 15 is divided into a consumables discarding part 151 and a liquid discarding part 152; the discarded consumables and the liquid are processed respectively. The amount of discarded objects contained in the discarded object placement structure 15 is equal to the space required to accommodate one replacement of the reagent consumables, which can be replaced together with the reagent consumables, which can ultimately reduce the difficulty of monitoring and increase the efficiency.

The enriched sample collection structure 18 sets the EP tube consumables placement structure 14, or the EP tube consumables placement structure 14 is set to the nearest other reaction zone; when the last step is performed, the EP tube is moved to the rich by the tube body moving structure 21. Collect the sample collection structure 18 domain, and then transfer the enriched supernatant to the EP tube through the liquid extraction and addition structure 22 for collection, and then perform detection.

Example 4

A process of using the device in Example 3 to carry out blood neuron-derived exosomes; four reaction areas are set; the sample tube enters the sample placement structure 16, and is moved by the tube body moving structure 21 to the reaction area 1; Shake, incubate, and centrifuge at least once, and the final process is centrifugation; add the corresponding reagents when needed; then complete at least one shake, incubation, and centrifugation by moving the sample tube or extracting the supernatant to the next reaction area , the corresponding reagents are added during the reaction, and the final process is the centrifugation process; when all the shaking incubation and centrifugation processes are completed, the sample tube or supernatant is moved to the final enrichment sample collection structure 18 for the next step of detection.

Referring to FIG. 5, the schematic diagram of FIG. 5 is only a schematic diagram of a process, and this process will involve various steps of exosome enrichment.

The descriptions of the above-mentioned embodiments are only for understanding the present invention. It should be pointed out that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements can also be made to the present invention, and these improvements will also fall within the protection scope of the claims of the present invention.

Claims

An automated device for effectively enriching neurogenic exosomes in blood, comprising a reaction rack at the bottom and a movable operating rack above; and a control for controlling the operating rack to run according to a predetermined track and the reaction rack to respond according to a set program It is characterized in that the reaction rack is provided with a shaking incubation structure, a centrifugal structure, a reagent placement structure, a consumables placement structure, a discard placement structure, a sample placement structure, a sample information identification structure, a reaction After the enrichment sample collection structure; the operation rack is provided with a tube body moving structure that realizes the movement of the tube body between different structures, and realizes the liquid taking and adding liquid structure for liquid transfer in the whole process; in addition, on the reaction rack or the operation rack An EP tube marking and identification structure is provided when the EP tube is used for the first time; the device also includes a storage module capable of storing information, and the control structure controls the oscillating speed and amplitude of the oscillating incubation structure, and controls the temperature required for the corresponding steps of the oscillating incubation structure; The control structure controls and monitors the amount of reagents in the reagent placement structure, and prompts for replacement when there is no reagent; the control structure also controls the centrifugal parameters of the centrifugal structure; the control structure monitors and records the information of the samples in the placement holes corresponding to the sample placement structure; the control structure controls the sample The information identifies the sample tube information and stores it in the storage module; the control structure controls the movement of the liquid taking and adding tube to perform the process of taking liquid and adding liquid, and the control structure controls the movement of the tube body. The structure grabs the corresponding tube body and moves to the corresponding next structure. In the area, the control structure controls the EP tube marking and identification structure to scan or identify the information on the EP tube and correspond to the corresponding sample tube information; avoid confusion or loss of patient information during the movement process; the control structure controls the last EP tube to be set to The enrichment sample was collected in the structure.
The automated device for effectively enriching neurogenic exosomes in blood according to claim 1, wherein a plurality of shaking incubation structures, centrifugation structures, reagent placement structures, consumables placement structures, and discards need to be set up according to the operation steps. Place the structure.
The automated device for effectively enriching neurogenic exosomes in blood according to claim 2, wherein when the sample or EP tube is in the stage of the shaking incubation structure, if it is necessary to add reagents, the corresponding shaking incubation structure There are corresponding reagent placement structures, consumables placement structures, and discard placement structures nearby.
The automatic device for effectively enriching neurogenic exosomes in blood according to claim 1, wherein, when only one batch of sample enrichment is performed in one operation, when the structure and shape of the sample tube and the EP tube are the same , set up a shaking incubation structure and a centrifugal structure in the whole process; or, if only one batch of sample enrichment is performed in one operation, when the shape of the sample tube and the EP tube are different, set the minimum number of tubes according to the requirements of different shapes. Objective To study the structure of shaking incubation and centrifugation.
The automatic device for effectively enriching neurogenic exosomes in blood according to claim 4, characterized in that, for the case where only one batch of samples is enriched in one process, a reagent placement structure, a consumables placement structure, Discard placement structure, set all reagent placement structures, consumables placement structures, and discard placement structures for exosome enrichment in a unified manner.
The automatic device for effectively enriching neurogenic exosomes in blood according to claim 1, wherein the operation rack is arranged on the setting top, and the setting top is a setting where the relative position of the reaction rack above the reaction rack does not change On the top, a moving track is provided on the top to ensure that the operation rack can move above any structure and perform corresponding operations.
The automatic device for effectively enriching neurogenic exosomes in blood according to claim 6, wherein the operation rack is an integral rack, on which a liquid taking and adding liquid structure and a tube body moving structure are arranged, and the liquid taking The liquid adding structure and the pipe body moving structure are structures that can move up and down on the operation frame, so that one structure can play its own role.
The automatic device for effectively enriching neurogenic exosomes in blood according to claim 6, wherein there are two operating racks, one for taking liquid and adding liquid and one for moving the tube body; When acting, they are respectively moved to the top of the corresponding structure for operation. When one operation frame is working, the other operation frame is moved to a position where it will not interfere with the other operation frame through the moving track.
The automatic device for effectively enriching neurogenic exosomes in blood according to claim 1, wherein the same number of operating heads are set on the liquid taking and adding liquid structure and the tube body moving structure.
The automatic device for effectively enriching neurogenic exosomes in blood according to claim 9, wherein the reagent placement structure corresponds to a single reagent and the number of liquid intake ports equal to the number of operation heads of the liquid intake and liquid addition structure is equal, and The positional relationship of the liquid sampling port is consistent with the positional relationship of the operation head; and the consumables placement structure sets the same number of sampling ports as the number of operation heads of the liquid sampling and liquid adding structure for corresponding single consumables; and the positional relationship of the material sampling port and the position relationship of the operation head Consistent.
The automated device for effectively enriching neurogenic exosomes in blood according to claim 9, wherein the number of placement holes in a single row is an integer multiple of the number of operating heads; or, the number of rows of placement holes and the number of operating heads Consistent.
The automatic device for effectively enriching neurogenic exosomes in blood according to any one of claims 1-11, wherein a fault alarm structure is further set between each structure and the control structure, and when the corresponding fault alarms After the structure sends an alarm signal, it prompts its corresponding structure to send a fault, or the control structure is also connected to a display structure, and the display structure displays the corresponding alarm structure alarm, prompting the corresponding structure fault to be repaired.
The automatic device for effectively enriching neurogenic exosomes in blood according to claims 1-11, wherein the number of reactions corresponding to the amounts of any two different reagents in the reagent placement structure is equal, and when the number of times reaches After that, it is prompted or displayed on the display structure to replace it.
The automatic device for effectively enriching neurogenic exosomes in blood according to any one of claims 1-11, wherein the number of reactions corresponding to the amount of any two different consumables in the consumable placement structure is also equal , or the amount of each consumable is the same as the number of reagent reactions.
The automatic device for effectively enriching neurogenic exosomes in blood according to any one of claims 1-11, wherein the discard placement structure is divided into a consumables discarding part and a liquid discarding part.
The automated device for effectively enriching neurogenic exosomes in blood according to claim 15, wherein the amount of discarded objects contained in the discard placement structure is equal to the space required to accommodate one replacement of reagent consumables.
The automatic device for effectively enriching neurogenic exosomes in blood according to any one of claims 1 to 11, wherein the reagent placement structure, the consumables placement structure and the discard placement structure are arranged in the middle of the reaction rack with a plate , and the combination is set under the settings board.
The automatic device for effectively enriching neurogenic exosomes in blood according to any one of claims 1-11, wherein the EP tube marking and identification structure is arranged on the tube body moving structure, and the EP tube top is arranged differently It can identify the structure one by one; by scanning and identifying the EP tube on the top, and corresponding with the corresponding sample tube information, the purpose of information matching is achieved.
The automated device for effectively enriching neurogenic exosomes in blood according to any one of claims 1-5, 9-11, wherein the entire enrichment process includes multiple shaking incubation and centrifugation processes, and each process It includes a shaking incubation and centrifugation step; according to the number of shaking incubation and centrifugation, the reaction rack of the entire equipment is divided into a plurality of corresponding reaction areas.
The automated device for effectively enriching neurogenic exosomes in blood according to any one of claims 6-8, wherein the entire enrichment process comprises multiple shaking incubation and centrifugation processes, and each process includes a shaking incubation And centrifugation step; according to the number of shaking incubation and centrifugation process, the reaction rack of the whole equipment is divided into a plurality of corresponding reaction areas.
The automatic device for effectively enriching neurogenic exosomes in blood according to claim 19, wherein a set of operation racks with a liquid taking and adding liquid structure and a tube body moving structure are arranged above each reaction area .
The automatic device for effectively enriching neurogenic exosomes in blood according to claim 20, characterized in that the moving track of the operation frame is a track that can satisfy the movement of the operation frame to achieve liquid extraction and gripping of the tube body.
The automated device for effectively enriching neurogenic exosomes in blood according to claim 20, wherein the moving track is also arranged in different regions, so as to be able to move in this region and obtain samples or EP tubes in the upper region movement track.
The automatic device for effectively enriching neurogenic exosomes in blood according to any one of claims 1-23, wherein the enrichment sample collection structure is provided with an EP tube consumables placement structure, or an EP tube consumables placement structure Set it to the nearest other reaction area; in the last step, move the EP tube to the enrichment sample collection domain through the tube body moving structure, and then move the enriched supernatant to the liquid sampling and adding structure. Collected in EP tubes and then detected.
Application of an automated device for effectively enriching neurogenic exosomes in blood in the enrichment of exosomes in blood.
A method for enriching exosomes in blood using the device of claim 20, characterized in that the sample tube enters the sample placement structure, and is moved from the tube body moving structure to the reaction area 1; at least one oscillation, incubation, The final process is the centrifugation process; the corresponding reagents are added when the reagents need to be added; then at least one shaking, incubation, and centrifugation process is completed by moving the sample tube or extracting the supernatant to the next reaction area, and adding the corresponding reagents during the process The reagents are reacted, and the final process is the centrifugation process; when all the shaking incubation and centrifugation processes are completed, the sample tube or supernatant is moved to the final enrichment sample collection structure for the next step of detection.