WO2023202351A1 - 一种按序加样装置及自动加样系统 - Google Patents

一种按序加样装置及自动加样系统 Download PDF

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Publication number
WO2023202351A1
WO2023202351A1 PCT/CN2023/085398 CN2023085398W WO2023202351A1 WO 2023202351 A1 WO2023202351 A1 WO 2023202351A1 CN 2023085398 W CN2023085398 W CN 2023085398W WO 2023202351 A1 WO2023202351 A1 WO 2023202351A1
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Prior art keywords
liquid outlet
piston
sequential
solution
liquid
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PCT/CN2023/085398
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English (en)
French (fr)
Inventor
宋世平
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浙江臻脉医疗科技有限公司
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Publication of WO2023202351A1 publication Critical patent/WO2023202351A1/zh

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/08—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/08—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
    • G01N35/085—Flow Injection Analysis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N35/1009—Characterised by arrangements for controlling the aspiration or dispense of liquids
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N35/1095—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices for supplying the samples to flow-through analysers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N2035/1027—General features of the devices

Definitions

  • the invention relates to the technical field of solution sampling devices, and in particular to a sequential sampling device and an automatic sampling system.
  • fully automatic experimental technology and equipment is one of the ways to solve human errors.
  • the fully automatic biochemical analyzer and fully automatic immune analyzer developed in the field of laboratory medicine make medical testing faster and the results more accurate.
  • fully automatic experimental equipment contains several mechanical modules necessary for automated design, making it large in size and weight, making it unsuitable for use in certain experimental scenarios (such as on-site inspections); at the same time, its batch experiment capabilities are not suitable for single experiments. operation, otherwise it will cause huge waste.
  • fully automatic equipment often requires a large amount of solution and is not suitable for micro-scale experimental operations.
  • Microfluidic chip also known as "lab on chip” is a new technology that integrates multiple experimental steps on a small-volume device.
  • the microfluidic chip itself for performing experiments is small in size, it requires external solution input and auxiliary operation equipment, including equipment to flow the solution (such as syringe pumps, peristaltic pumps or centrifuges, etc.), solution switching devices (microvalves, for example) bubbles, and corresponding pipelines) and flow rate control equipment, etc.
  • one method is to manually replace different solutions at the solution entry end, pump them in with a syringe pump or peristaltic pump, or inhale through the negative pressure caused by the end centrifuge. Due to the replacement of solutions, Manual operation is required, and bubbles will inevitably be introduced during the replacement process; one method is to set up an automatic switching device at the solution inlet end, such as using an automatically controlled fluid multi-way valve to automatically switch one liquid to another. This method is also difficult to completely avoid the introduction of bubbles.
  • the internal volume of the microfluidic chip structure is very small, which is mainly suitable for microliter-level liquid experiments, but not suitable for milliliter-level liquid experiments.
  • the CN205163822U discloses a prefilled segmented syringe, which can prefill solutions in different segments to achieve sequential addition of samples.
  • the syringe includes a syringe needle head and a syringe needle tube part.
  • the syringe needle tube part includes a number of primary needle tubes, secondary needle tubes and multi-stage needle tubes that are interconnected up and down.
  • the bottom of the secondary needle tube and multi-stage needle tubes are Both are equipped with a piston that can make up and down piston movements together with the needle tube of the current level in the needle tube of the next level.
  • the piston at the bottom of the secondary needle tube and the multi-stage needle tube is equipped with a sealing device.
  • the secondary needle tube and the multi-stage needle tube are equipped with a sealing device.
  • the inner wall of the needle tube is provided with a hole corresponding to the outer wall of the upper level needle tube.
  • Matching locking mechanism; the syringe needle head includes an injection needle, and the injection needle is penetrated at the bottom of the primary needle tube; the sealing device is arranged above the vertical line of the injection needle, and the medicine in the bottommost needle tube After the injection is completed, the injection needle passing through the bottommost needle tube pierces the sealing device at the bottom of the previous needle tube, and then the medicine in the needle tube can be injected, and then the above operation can be repeated to achieve the technical effect of segmented advancement.
  • this solution only has one outlet, when it is necessary to automatically add samples to different locations in sequence, it cannot be completed through only a single outlet.
  • the present invention provides a sequential sample adding device and an automatic sample adding system to solve at least one of the above technical problems.
  • the technical solution of the present invention is: a sequential sampling device, which includes a sampling tube.
  • the sampling tube includes a tube wall. N storage spaces are provided at one end toward the other end of the sampling tube, and any one of the storage spaces is It includes a solution section pre-installed in the tube and a piston at its rear, with N greater than 1; the tube wall is provided with a first liquid outlet and N-1 second outlets that can communicate with the solution section in the first storage space. Liquid port; the first liquid outlet is pre-sealed; the N-1 second liquid outlets are respectively arranged in the area closed by the piston of the previous storage space, and the piston serves as a partition between the liquid outlet and the liquid outlet.
  • the piston open to the outside is used to contact the pushable power device.
  • the pre-sealed first liquid outlet is opened, and the corresponding contact is pushed by the power device.
  • the piston releases the solution segment in the first storage space, and the piston in the previous storage space moves to a position where the latter solution segment is connected to the second liquid outlet, so that when the latter solution segment is pushed, it can pass through the corresponding second outlet.
  • the liquid port is released.
  • the solution cannot flow out due to external air pressure without pushing.
  • the pre-sealing of the first liquid outlet means that the solution section of the first storage space cannot communicate with the outside world through the first liquid outlet.
  • the method of preloading the liquid solution into the sample tube may be to open a hole on the outside of the tube wall, inject the solution through the hole, and then close the opening.
  • the power device can be a human hand, or it can be a pneumatic drive, a hydraulic drive, or a motor drive, as long as it can push the piston to move. Through the piston movement, the piston can be released from the sample addition port according to the requirements for sequential sample addition. liquid solution. If the above-mentioned pneumatic drive, hydraulic drive, and motor drive device are used as the power device, the time interval, propulsion speed, etc. of the power device drive can be controlled by the controller to realize that the solution required for the reaction is automatically added in the set sequence and time interval. Release.
  • the power device is preferably a motor drive device, including a piston rod and a driver. The other end of the piston rod is connected to a driver capable of linear motion. The driver can push the piston rod to drive the piston to move in the direction of its corresponding solution section.
  • liquid solutions are pre-installed in the sampling tube.
  • the pre-installed liquid solutions can be automatically propelled by the power device, which can greatly Improving the simplicity of the reaction procedure is suitable for making portable detection equipment, and is more suitable for on-site detection application scenarios; if the pre-installed liquid solutions are different solutions, the design of the present invention can allow two or more different solutions to be added completely independently, without There is pipeline contamination.
  • the sample adding tube is a whole tube, and N consecutive storage spaces are arranged in sequence from one end to the other end of the whole tube, and the first liquid outlet and the N-1th The two liquid outlets are arranged at separate intervals; all the second liquid outlets are located under the piston in the previous storage space; the end of the piston at the end that is open to the outside is used to contact the pushable power device.
  • the above scheme uses a pipeline, and the piston divides the pipeline into two or more storage spaces, and each storage space is in a serial relationship.
  • the first liquid outlet is in a pre-sealed state.
  • the method of pre-sealing can be external air pressure sealing, valve sealing, plugging, plugging and other sealing covers, or the first liquid outlet can be covered with other removable In the seal (such as a movable piston); the other N-1 second liquid outlets are in the area closed by the piston in the previous storage space and are not connected to the solution in the subsequent storage space in the initial state.
  • sequential sample addition begins, first unblock the first liquid outlet so that the solution section in the first storage space can communicate with the outside world.
  • the piston is pushed by the power device, and the solution section in the first storage space is released through the first liquid outlet.
  • the second liquid outlet covered under it is exposed to the solution in the latter storage space, so that when the power device continues to push, the corresponding solution in the latter storage space can be discharged from the connected
  • the second liquid outlet is released.
  • Each liquid outlet is exposed in sequence, so that two or more liquid solutions required for the reaction can be automatically added to the set area in the set order and time interval, which can greatly improve the degree of automation of the reaction.
  • the sampling tube includes N independent cavities arranged side by side, and N storage spaces are respectively located in N independent cavities; each independent cavity includes a tube wall and a solution segment and a piston within it. , the pipe wall of the first independent cavity is provided with a first liquid outlet connected to the solution section, and the pipe walls of the other N-1 independent cavities are provided with a second communication port connected to the solution section.
  • the first liquid outlet The port is pre-sealed; each independent cavity also includes a first communication port covered under the piston and close to one end of the solution section and a second liquid outlet located adjacent to it.
  • the opening and the second liquid outlet are in the same position, the first communication port and the second liquid outlet are connected through the groove, so that the second liquid outlet is connected to the solution section of the subsequent independent cavity.
  • each piston that is open to the outside can be used to contact the power device, and each power device pushes the corresponding piston respectively.
  • This kind of independent cavities arranged side by side has a more reasonable overall length. If the front end of the power unit is a piston rod, the technical problem of the piston rod extending out of the sample tube too long in serial mode can be avoided.
  • the driver of each piston can be independently The driver can also be linked through programming, so that high-throughput sequential automated sample addition can be achieved.
  • each independent cavity is a relatively independent reaction unit, if necessary, the communication pipe between the two independent cavities can be cut off, so that the second communication port of the latter independent cavity can become a new
  • the first liquid outlet allows the independent chambers arranged side by side to be split into multiple sequential sampling devices, which helps the operator to handle flexibly.
  • the first liquid outlet and the N-1 second liquid outlets can be connected to liquid outlet pipes respectively.
  • This independent setting of the liquid outlet pipes ensures that different solutions There is no mutual contamination between them, and it also helps to guide the liquid outlet tubes to the sample inlets at different locations to meet the needs of adding different liquids at different sample locations. For example, some microfluidic structure chips or chromatographic structure chips can be added at different locations. Requirements for adding different liquids.
  • a retention chamber for accommodating the solution is provided in the middle of the liquid outlet pipe.
  • the outlet of the liquid outlet pipe is arranged corresponding to the inlet of the corresponding solution receiving end.
  • the arrangement of the retention cavity can avoid the capillary effect at the solution receiving end and the formation of siphon phenomenon. .
  • the invention also discloses a sequential automatic sample adding system, which includes the aforementioned sequential sample adding device, a piston rod, and a driver.
  • the driver is preferably driven by a stepper motor.
  • the outlet of the stepper motor is connected to a ball screw.
  • the ball screw A screw nut is provided on the top, and the end face of the screw nut is in contact with the piston rod;
  • the stepper motor is connected to the controller, and the controller controls the screw nut to intermittently push the piston rod movement through a delay.
  • the movement of the stepper motor is controlled by the controller.
  • the controller control content includes the time interval and propulsion speed of the propulsion start.
  • the solution required for the reaction is automatically added to the set area in the set order and time interval, which greatly improves the degree of automation of the reaction. , the liquid in different parts is released at a set time through the piston rod, eliminating manual operation errors and greatly improving the accuracy of the reaction results.
  • the controller may be a programmable controller PLC, for example.
  • the sequential sample adding device of the present invention can be matched with a variety of scenarios requiring sequential sample addition, and can be pushed manually or automatically. For example, it can be matched with a detection chip with a chromatographic structure or a microfluidic structure that requires sequential addition of samples. Therefore, the present invention also provides an automatic detection system for sequential sample addition, including a sequential sample addition device, a power device, a detection device, the first liquid outlet of the sequential sample addition device and the N-1 The second liquid outlets are respectively connected to different sample addition inlets of the detection device, and the power device automatically drives the piston of the sequential sample addition device to release the liquid in sequence.
  • the detection device is a detection chip with a chromatographic structure or a microfluidic structure.
  • chromatographic electrochemical biochip Take the detection chip with a chromatographic structure as an example. Some detection chips with a chromatographic structure need to add samples multiple times in sequence, such as chromatographic electrochemical biochips for pesticide detection and immunoassays. Chromatographic electrochemical biochip. Take the chromatographic electrochemical biochip for immunodetection of AIDS antibodies (HIVAb) as an example.
  • the HIV antigen (S-HIVAg) is fixed on the working electrode, which is the detection area, and the front end of the chromatographic test strip is sprayed on it.
  • the HIV antigen-horseradish peroxidase (HRP) complex (HIVAg-HRP) is adsorbed, and the HIVAg-HRP spray area accepts the HIV antibody (HIVAb) to be tested and the washing solution; the detection at the working electrode is Zone accepts substrate TMB solution.
  • the HIV antibody sample needs to be added by yourself, and then the washing solution and substrate TMB solution need to be added in sequence, with an interval of ten minutes.
  • the washing liquid can pass through Through chromatography, other substances that are not bound to the HIV antigen (S-HIVAg) in the detection area are fully eluted, causing them to move out of the detection area, leaving "Ag-Ab-Ag (S-HIVAg-HIVAb-HIVAg) in the detection area.
  • -HRP -HRP
  • microfluidic electrochemical biochips for pesticide detection and immunoassays.
  • Microfluidic electrochemical biochip for detection Taking the microfluidic electrochemical biochip for immunological detection of AIDS antibodies (HIVAb) as an example, the HIV antigen (S-HIVAg) is fixed on the working electrode, which is the detection area. Among them, HIV antibody (HIVAb) needs to be added by itself. After that, the primary washing solution, HIV antigen-enzyme complex (HIVAg-HRP), secondary washing solution and TMB substrate solution need to be added in sequence, with an interval of ten minutes between each. .
  • the primary washing liquid fully washes other substances that are not bound to the HIV antigen (S-HIVAg) in the detection area through the power microflow, causing them to move out of the detection area; then, the HIV antigen-enzyme complex enters the detection area through the power microflow and interacts with the detection area.
  • the captured HIV antibody binds to the detection area, forming an "Ag-Ab-Ag (S-HIVAg-HIVAb-HIVAg-HRP)" type sandwich complex in the detection area; the secondary washing liquid fully elutes the uncaptured HIV antibodies through dynamic microflow. HIV antigen-enzyme complex; the subsequently added TMB substrate and HRP undergo an enzymatic reaction to output a current signal.
  • Figure 1 is a schematic diagram of the internal installation structure of Embodiment 1 of the present invention.
  • Figure 2 is an enlarged view of part A of Figure 1.
  • Figure 3 is a usage state change diagram of Embodiment 1 of the present invention.
  • FIG. 4 is a diagram of usage state changes according to Embodiment 2 of the present invention.
  • Figure 5 is an enlarged view of part B of Figure 4.
  • Figure 6 is an enlarged view of part C of Figure 4.
  • Figure 7 is a schematic structural diagram of the liquid receiving pipe in Embodiment 4 of the present invention.
  • Figure 8 is a structural diagram of a chromatographic electrochemical biochip used in Embodiment 1 of the present invention.
  • Figure 9 is a graph showing the results of testing using a chromatographic electrochemical biochip according to Embodiment 1 of the present invention.
  • the sampling tube 1 is a whole tube, including the tube wall.
  • Four storage spaces are provided at one end of the sampling tube 1 toward the other end. Any one of the storage spaces Each space includes a solution section 4 pre-installed in the tube and a piston 5 at its rear; the tube wall is provided with a first liquid outlet 2 and three second liquid outlets 3 that can communicate with the solution inside it;
  • the first liquid outlet 2 is pre-blocked by a plug (the plug is not shown in the figure); the three second liquid outlets 3 are respectively arranged in the area closed by the piston 5 of the previous storage space, and the piston 5 It plays the role of switching and promoting the partition and liquid outlet.
  • the rear end of the fourth piston is open and provides access to the power unit.
  • the method of preloading the liquid solution into the sample tube is to open a hole in the upper part of the tube wall, inject the solution through the hole, and then close the opening (not shown in the figure).
  • the lower end of the second liquid outlet on the tube wall is open to the outside, but at the initial time, its upper end is within the coverage of the piston, so any second liquid outlet None of them can connect to the solution section of the latter storage space.
  • Figure 3 shows the use process of the serial sequential sampling device.
  • the end of the piston 5 in the fourth storage space that is open to the outside contacts the piston rod 6.
  • the piston rod 6 can be pushed manually or driven by a stepper motor (the stepper motor is not shown in the figure).
  • the pre-sealed first liquid outlet 2 is opened, and the contacting piston 5 is pushed through the piston rod 6.
  • the solution in the tube and the piston as a whole move forward, and the solution section 4 in the first storage space is moved from The first liquid outlet 2 is released, and the piston 5 of the first storage space moves forward to expose the second liquid outlet 3 to the solution section 4 of the second storage space.
  • the pushing stops stops.
  • the solution in this solution section will be released from the second liquid outlet 3 when pushing is resumed.
  • the piston 5 of the previous storage space moves to a position where the next solution section 4 is connected to the second liquid outlet 3, so that when the latter solution section 4 is pushed, it can be released through the second liquid outlet 3.
  • the solution in the solution section can be released in sequence, thereby achieving the effect of sequential addition. If a stepper motor is used for driving, the outlet of the stepper motor is connected to the ball screw, and a screw nut is provided on the ball screw.
  • the end face of the screw nut is in contact with the piston rod; the stepper motor is connected to the controller, and the controller passes The delay controls the screw nut to intermittently push the piston rod movement.
  • threads or buckles can be provided outside the liquid outlet to facilitate tight connection with the liquid outlet pipe.
  • This embodiment shows a sequential sampling device containing four storage spaces.
  • the storage spaces can be changed between two or more according to actual needs.
  • Two or more liquid solutions can be the same or different. Especially when the solutions in the solution section are different, different liquid outlets can be connected separately, which can not only avoid contamination between solutions, but also lead different solutions to different places, avoid additional complex pipelines, and only use the piston By pushing, you can automatically add samples in different areas in sequence.
  • a sequential sampling device is in side-by-side mode.
  • the sampling tube 1 includes five independent cavities arranged side by side, and the five storage spaces are respectively located in the five independent cavities.
  • Each The independent cavity includes the tube wall and the solution section 4 and piston 5 inside.
  • This embodiment adopts independent cavities arranged side by side.
  • the overall length is more reasonable and avoids the need for the piston rod to extend when there is more storage space in the serial mode.
  • Technical problem of too long sample tube length Specifically, a first liquid outlet 2 connected to the solution section 4 is provided on the wall of the first independent cavity, and a second communication port 82 connected to the front end of the solution section 4 is provided on the pipe walls of the other four independent cavities.
  • the first to fourth independent cavities also include a first communication port 81 covered under the piston 5 and close to the end of the solution section 4 and a second liquid outlet 3 located adjacent to it.
  • the second liquid outlet 3 and the The first communication port 81 is not connected in the initial state;
  • the second communication port 82 on the wall of the second to fifth independent cavities is connected to the first communication port 81 of the previous independent cavity through the communication tube 8;
  • the first to fourth independent cavities are connected to each other through the communication tube 8.
  • the piston 5 of the cavity is provided with a groove 51 arranged along the axial direction on the outer ring at one end away from the solution section 4.
  • the length of the groove 51 can cover the first communication port 81 and the second liquid outlet 3.
  • the first liquid outlet 2 is pre-sealed in its initial state so that it can be unsealed during use.
  • One end of the piston 5 in any of the independent chambers that is open to the outside is used to contact the respectively provided piston rod 6, and each piston rod 6 can be connected to a corresponding driver.
  • the first liquid outlet 2 is connected with the first liquid outlet pipe 7 so as to be guided to the area where sample is required.
  • a valve (not shown in the figure) can be used to connect the first liquid outlet pipe 7 and achieve pre-sealing.
  • Figure 4 shows the use process of the side-by-side sequential sampling device.
  • the valve When it is necessary to add samples in sequence, the valve is rotated, the first liquid outlet 2 is connected to the first liquid outlet pipe 7, the piston rod 6 of the first independent chamber is driven, and the piston 5 is pushed to push the solution section 4 at its front end through The first liquid outlet 2 and the first liquid outlet pipe 7 are released outward.
  • the groove 51 on the piston 5 moves to a position covering the first communication port 81 and the second liquid outlet 3, so that The first communication port 81 and the second liquid outlet 3 are connected through the groove 51 through the connecting pipe 8 and the solution section 4 of the latter independent cavity; when the piston 5 in the latter independent cavity is pushed by its corresponding piston rod 6, the solution sequence in the corresponding solution section 4 It is released through the second communication port 82 and communication pipe 8 of the latter independent cavity, the first communication port 81 of the previous storage space, the groove 51 and the second liquid outlet 3 .
  • the second liquid outlet 3 is connected to the corresponding second liquid outlet pipe 9 to guide it to the area where sample is required.
  • a sequential automatic sample adding system includes a sequential sample adding device, a piston rod 6, and a driver.
  • the driver includes a stepper motor, an outlet of the stepper motor, and a ball.
  • the screw is connected, and the ball screw is provided with a screw nut, and the end face of the screw nut is in contact with the piston rod 6;
  • the stepper motor is connected to the controller, and the controller controls the screw nut to intermittently push the movement of the piston rod 6 through a delay.
  • the present invention controls the movement of the stepper motor through a controller.
  • the controller control content includes the time interval and propulsion speed of the propulsion start, so that the solution required for the reaction is automatically added to the set area in the set order and time interval, which greatly improves the efficiency of the stepper motor. It improves the degree of automation of the reaction, and uses the piston rod to release liquids in different parts at a set time, eliminating manual operation errors and greatly improving the accuracy of the reaction results.
  • the screw nut can be connected and fixed with the piston rod 6 to prevent the piston rod from moving accidentally and driving the piston.
  • Embodiment 4 is a diagrammatic representation of Embodiment 4:
  • the first liquid outlet 2 and the second liquid outlet 3 can be connected to liquid pipes respectively.
  • a retention chamber 12 for accommodating the solution is provided in the middle of the liquid pipe.
  • the outlet of the liquid pipe is connected to the corresponding solution receiving end inlet. Corresponding arrangement.
  • HIV antigen (HIVAg) (Guangdong Feipeng Biological Co., Ltd.); horseradish peroxidase (HRP)-labeled HIV antigen (HIVAg-HRP) (Guangdong Feipeng Biological Co., Ltd.); HIV-I type antibody (HIV-IAb )(5k)(Guangdong Feipeng Biological Co., Ltd.); Bovine serum albumin (BSA) (Sangon Bioengineering Shanghai Co., Ltd.); Casein (sigma Company); 3,3′,5,5′ - Tetramethylbenzidine (TMB) substrate solution (0.4g/L) (Thermo Company); 1*PBS buffer solution; 1*PBST washing solution (500 ⁇ L Tween-20 was added to 1L of 1*PBS buffer) ; pH 9.6 sodium carbonate buffer solution (coating solution).
  • HRP horseradish peroxidase
  • HRP horseradish peroxidase
  • HIV antigen HIV antigen
  • HRP horseradish
  • the experimental detection structure includes a chromatographic electrochemical detection chip, where A is the front end of the chip; B is the back end of the chip.
  • An upper cover 32 is provided above the chromatographic electrochemical detection chip.
  • a lower cover 31 is provided below, and a rectangular opening slot is provided on the mating surfaces of the upper cover 32 and the lower cover 31.
  • the electrochemical detection chip is installed in the opening slot.
  • the openings of the opening slot are all front openings 33.
  • the upper cover The top surface of 32 is provided with a sampling hole 322 for HIV I antibody sampling and washing solution release, and a sampling hole 321 for substrate release in sequence from front to back;
  • the chromatographic electrochemical detection chip is composed of an electrode chip and a tightly adhered It consists of a chromatography test strip on it, where the electrode chip includes a working electrode 11 and a counter electrode and a reference electrode located on both sides of the working electrode 11.
  • the three are produced on the substrate through a screen printing process to form an electrode chip.
  • the leads of the three electrodes are assembled at the front end of the chip, and the leads extend from the front end of the test strip.
  • HIV antigen (S-HIVAg) is fixed on the working electrode 11;
  • the chromatography test strip is composed of a sample pad 21, a chromatography membrane 22, and a water-absorbent pad 23 sequentially stacked.
  • the sample pad 21 is provided with HIVAg- HRP spraying area 24, the detection area 25 is located in the middle of the sample pad 21, and the working electrode 11 is located within the range of the detection area 25;
  • HIV antigen HIVAg
  • 1*PBST washing solution and HRP enzyme catalytic substrate TMB solution are pre-packaged in the sampling tube 1 of the serial sequential sampling device similar to Example 1; refer to Figure 1, the solution segment contained in the first storage space 4 is 150 ⁇ L of 1*PBST washing solution, followed by the first section of piston 5.
  • the solution section 4 contained in the second storage space is 100 ⁇ L of substrate TMB solution, followed by the second section of piston 5, and the second section of piston 5.
  • the end is in contact with the piston rod 6 for pushing, and the screw that pushes the piston rod moves is driven by a programmable stepper motor.
  • the first liquid outlet of the sample adding tube 1 corresponds to the sample adding hole 322 of the chip; the second liquid outlet corresponds to the sample adding hole 321 of the chip.
  • the HIV-I antibody with a titer of 5k was diluted in different multiples with the above blocking solution (1:1000, 1:500, 1:100, 1: 20), take 150 ⁇ L and drop it at the injection hole 322 to form a conjugate of HIV-I antibody and HIVAg-HRP immunoreaction, and move it to the detection area through chromatography, forming " Ag-Ab-Ag(S-HIVAg—HIV-I Ab—HIVAg-HRP)” type sandwich conjugate;
  • the pre-sealing of the sample addition tube is released, and the stepper motor is controlled by the program to drive the piston 5 by pushing the piston rod, so that the encapsulated 1*PBST washing liquid is released through the first liquid outlet 2.
  • the sample hole 322 is released to the test strip of the chromatographic electrochemical detection chip.
  • the piston rod is pushed again by program control to drive the piston.
  • the encapsulated substrate TMB solution is released from the second outlet hole, and is released to the test strip detection area 25 corresponding to the working electrode 11 of the chromatographic electrochemical detection chip through the sampling hole 321.
  • the above-mentioned 10-min interval is for the washing solution to fully elute other substances that are not bound to the HIV antigen (S-HIVAg) in the detection area through chromatography, so that they can move out of the detection area; the substrates TMB and HRP added later will generate enzymes.
  • the current data is collected through the time current curve (i-t); the test potential is -0.1V, and the current signal value after the test time of 50s is the output detection data.
  • test data of HIV-free blocking solution is blank background data.
  • HIV-I antibody was diluted 1:1000, 1:500, 1:100, 1:20.
  • the detection results are shown in Figure 9. Even if the antibody is diluted 1:1000 times, it can be clearly distinguished from the background signal.
  • serial mode sequential liquid adding structure and the side-by-side mode sequential liquid adding structure of the present invention can be flexibly selected for sequential liquid adding, which can make the portable device adaptable to a variety of application scenarios.

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Abstract

一种按序加样装置及自动加样系统，按序加样装置包括加样管（1），加样管（1）内一端向着另一端设置有N个存储空间，每个存储空间均包括溶液段（4）和活塞（5），活塞（5）可以与活塞杆（6）接触以被推动；加样管（1）管壁上设置有可与加样管（1）内部溶液相通的第一出液口（2）、N-1个第二出液口（3），N-1个第二出液口（3）分别布置在前一个存储空间活塞（5）封闭的区域内；活塞杆（6）推动活塞（5），前一存储空间溶液段（4）的溶液加完，后一溶液段（4）的第二出液口（3）露出，溶液在无推动的情况下不能流出，达到按序按需加样的目的。在加样管（1）中预装二个及其以上的液体溶液，适合于制作便携式器件及现场检测应用场景；通过驱动器推动活塞杆（6）运动，能根据按序加样的要求分别从加样口释出液体溶液。

Description

一种按序加样装置及自动加样系统 技术领域
本发明涉及溶液加样装置技术领域，具体涉及一种按序加样装置及自动加样系统。
背景技术
在各种化学和生物实验中，人为操作误差会造成结果的重大差异，所以如何在实验操作中减少人为误差是相关技术发展的推动力之一。涉及溶液的化学和生物实验流程往往包含多个液体按序加入程序，时间和液体加入量的精确控制对操作人员要求更高，也更容易在操作中产生人为误差。
全自动实验技术和设备的发展是解决人为误差的路径之一，如检验医学领域发展起来的全自动生化分析仪和全自动免疫分析仪等使医学检验更加快速，结果更加准确。但全自动实验设备因包含若干自动化设计所必须的机械模块，而使其体积和重量较大，不适于某些实验场景(如现场检验)下使用；同时，其批量实验能力不适于单次实验操作，否则造成巨大浪费。此外全自动设备往往需要较多溶液量，不适于微量实验操作。
基于微流控结构的芯片技术适于微量操作，但在自动化方面面临较多困难，特别是在按序加样的情况下。微流控芯片也称“芯片实验室”，是将多个实验步骤集成在小体积器件上的新技术。虽然执行实验的微流控芯片本身体积较小，但需要外部溶液输入和辅助运行设备，包括使溶液流动的设备(如注射泵、蠕动泵或离心机等)、溶液切换装置(微阀，例如气泡，和相应管路)和流速控制设备等。微流控芯片若需实现按序加样，一种方法是在溶液进入端手动更换不同溶液，由注射泵或蠕动泵将其泵入，或通过末端离心机造成的负压吸入，由于更换溶液需要手动操作，更换过程中不可避免地会引入气泡；一种方法是在溶液进入端设置一自动切换装置，例如采用自动控制的流体多通阀，将一种液体自动切换到另一种液体，该方法也难以完全避免气泡引入。此外，微流控芯片结构内部容积很小，主要适合微升量级液体实验，不适于毫升以上量级液体实验。
因此，在需要液体多次按顺序加样的操作流程中，对于自动按序精确加样的需求仍亟待满足。
CN205163822U中公开了一种预充式的分段式注射器，可以在不同的段中预充溶液，实现按序加样。该注射器包括注射器针头部和注射器针管部，所述的注射器针管部包括若干个上下相互套接而成的一级针管、二级针管及多级针管，所述的二级针管及多级针管底部均设有能和本级针管一起在下一级针管内做上下活塞运动的活塞，所述的二级针管及多级针管底部的活塞上均设有密封装置，所述的二级针管及多级针管的内壁设有和上一级针管外壁相 配合的锁定机构；所述的注射器针头部包括注射针，所述的注射针穿设在一级针管的底部；所述的密封装置设置在注射针的垂直线上方，在最底部针管内的药品注射完成后，穿设在最底部的针管的注射针将上一针管底部的密封装置刺破，则可以注射该针管内的药品，然后重复上面的操作，以实现分段式推进的技术效果。但由于该方案仅存在一个出口，在需要自动按序向不同位置加样时，无法仅仅通过单一出口完成，后续仍需要另外的自动控制的流体多通路阀管路配合；同时，在分段推进的基础上按序释出不同溶液从而实现按序加样时，出口处共用一个通道，会产生溶液污染问题。
因此，如何将两个及其以上的液体溶液分别封装于一个装置的多个部位中并根据按序加样的要求分别释出，如何通过控制按序加样装置推进部件的运动使不同部位的液体在设定的时间进行释放，仍是实践中急需解决的问题。
发明内容
针对现有技术存在的问题，本发明提供一种按序加样装置及自动加样系统，以解决上述至少一种技术问题。
本发明的技术方案是：一种按序加样装置，包括加样管，所述加样管包括管壁，所述加样管内一端向着另一端设置有N个存储空间，任意一个存储空间均包括预装于管内的溶液段及其后部的活塞，N大于1；所述管壁上设置有可与第一存储空间内溶液段相通的第一出液口、N-1个第二出液口；所述第一出液口预封闭；所述N-1个第二出液口分别设置在其前一个存储空间的活塞封闭的区域内，所述活塞起区隔、出液口的开关及推动作用；所述活塞敞开于外部的一端用于接触可推动的动力装置，当需要进行按序加样时，开放预封闭的所述第一出液口，通过动力装置推动对应接触的活塞，第一存储空间的溶液段释出，前一个存储空间的活塞运动至使后一溶液段与第二出液口连通的位置，使得后一溶液段被推动时可经由相应的第二出液口释出。溶液在无推动的情况下由于外气压的压力不能流出。所述的第一出液口预封闭是指第一存储空间的溶液段不能经由第一出液口与外界连通。
向加样管中预装液体溶液的方法可以是在管壁外部开孔，从孔中注入溶液，而后将该开孔封闭。
本发明使用时，动力装置可以是人手，也可以是气压驱动、液压驱动、电机驱动装置，只要能够推动活塞运动即可，通过活塞运动能根据按序加样的要求分别从加样口释出液体溶液。若采用上述气压驱动、液压驱动、电机驱动装置作为动力装置，可以通过控制器控制所述动力装置驱动的时间间隔、推进速度等，以实现反应所需加入的溶液按设定顺序和时间间隔自动释出。所述的动力装置优选电机驱动装置，包括活塞杆，驱动器，活塞杆的另一端与能做直线运动的驱动器连接，驱动器能推动活塞杆带动活塞向着其对应的溶液段方向运动。
本发明在加样管中预装两个及以上的液体溶液，检测时最多只需一次人工加样(人工加入待测实验样品)，预装的液体溶液均可通过动力装置自动推进，可大幅提升反应程序的简便程度，适合于制作便携式检测设备，更加适用于现场检测应用场景；若预装的液体溶液为不同溶液，本发明的设计可以使得两种以上的不同溶液完全独立加样，不存在管道污染。
优选的一种方案为：所述加样管为一根整管，整管内一端向着另一端依次设置有N个连续的存储空间，所述第一出液口和所述的N-1个第二出液口分别间距布置；所有第二出液口均分别位于前一个存储空间的活塞下方；最末端的活塞敞开于外部的一端用于接触可推动的动力装置。
上述方案采用一条管路，活塞将管路分割为两个及以上的存储空间，各存储空间为串行关系。初始状态时，第一出液口处于预封闭状态，预封闭的方法可以是外气压封闭、阀门封闭、栓塞、堵头等密封盖封闭，也可以将第一出液口覆盖于可移除的其他密封件(例如可移动的活塞)中；其他N-1个第二出液口处于前一个存储空间活塞封闭的区域，在初始状态时不与后一个存储空间中的溶液连通。按序加样开始时，首先解除第一出液口的封闭，使第一存储空间的溶液段与外界可连通，通过动力装置推动活塞，第一存储空间的溶液段通过第一出液口释出，待活塞被推动到特定位置时，其下覆盖的第二出液口暴露于后一个存储空间的溶液中，使得动力装置继续推动时可将对应的后一个存储空间的溶液从相连通的第二出液口释出。各出液口依次暴露，使得反应所需加入的二个及其以上的液体溶液可按设定顺序和时间间隔自动加入到设定区域，可大幅提升反应自动化程度。
优选的另一方案为：所述加样管包括N个并排布置的独立腔体，N个存储空间分别位于N个独立腔体内；每个独立腔体包括管壁及其内的溶液段和活塞，第一独立腔体的管壁上设置有连通溶液段的第一出液口，其他N-1个独立腔体的管壁上均设置有连通溶液段的第二连通口，第一出液口预封闭；每个独立腔体还包含覆盖在活塞之下的且靠近溶液段一端的第一连通口及其临近设置的第二出液口，第二出液口和第一连通口初始状态不连通；N-1个独立腔体管壁上的第二连通口通过连通管与前一独立腔体的第一连通口连通；每一个独立腔体的活塞在远离溶液段的一端外圈设置有沿着轴向布置的槽，所述的槽的长度能覆盖第一连通口与第二出液口；当活塞被推动时，所述的槽随之移动，当槽达到同时覆盖第一连通口与第二出液口的位置时，第一连通口与第二出液口通过槽相连通，使得第二出液口与其后一个独立腔体的溶液段连通。
上述方案中，每个活塞敞开于外部的一端均可用于与动力装置接触，每个动力装置分别推动相应的活塞。这种并排布置的独立腔体，整体长度更加合理。若动力装置前端为活塞杆，可避免串行模式时活塞杆伸出加样管长度过长的技术问题，每个活塞的驱动器可以单独 驱动，也可以通过编程进行联动，从而可以实现高通量按序自动化加样。
另外，由于每个独立腔体都是一个相对独立的反应单元，在需要的情况下，可以截断两个独立腔体之间的连通管，使得后一个独立腔体的第二连通口可以成为新的第一出液口，这可以使得并排布置的独立腔体可以分拆成多个按序加样装置，有助于操作方的灵活处理。
本发明的任何一种情况下，所述的第一出液口和所述的N-1个第二出液口都可以分别连接出液管，这种出液管的独立设置保证了不同溶液间不互相污染，也有助于将出液管引向不同位置的加样入口，实现不同加样处加不同液体的需求,例如实现一些微流控结构芯片或层析式结构芯片在不同加样处加不同液体的需求。
优选地，出液管中部设置有容纳溶液的滞留腔，出液管的出口与相应的溶液接纳端入口对应布置，滞留腔的设置可以避免溶液接纳端处出现毛细管效应，也可以避免虹吸现象形成。
本发明还公开一种按序自动加样系统，包括前述的按序加样装置，活塞杆，驱动器，其中驱动器优选采用步进电机驱动，步进电机的出口与滚珠丝杠连接，滚珠丝杠上设置有丝杠螺母，丝杠螺母的端面与活塞杆接触；步进电机与控制器连接，控制器通过延时控制丝杠螺母间歇推动活塞杆运动。通过控制器控制步进电机的运动，控制器控制内容包括推进启动的时间间隔、推进速度，反应所需加入的溶液按设定顺序和时间间隔自动加入到设定区域，大幅提升了反应自动化程度，通过活塞杆使不同部位的液体在设定的时间进行释放，排除了人工操作误差，大幅提升了反应结果的精确度。控制器例如可以是可编程控制器PLC。
本发明的按序加样装置可以与多种需要按序加样的场景匹配，可以手动推动，也可以自动推动。例如，与需要按序加样的层析式结构或微流控结构的检测芯片进行匹配。因此，本发明还提供一种按序加样自动检测系统，包括按序加样装置，动力装置，检测装置，所述按序加样装置的第一出液口和所述的N-1个第二出液口分别对接到检测装置的不同加样入口，所述动力装置自动驱动所述按序加样装置的活塞使得液体按序释出。优选地，检测装置为具有层析式结构或微流控结构的检测芯片。
以层析式结构的检测芯片为例，部分具有层析式结构的检测芯片的使用场景中需要多次按序加样，例如用于农药检测的层析式电化学生物芯片、用于免疫检测的层析式电化学生物芯片。以艾滋病抗体(HIVAb)免疫检测用层析式电化学生物芯片为例，其工作电极上固定了HIV抗原(S-HIVAg)，为检测区，其上的层析用试纸条的前端喷样吸附了HIV抗原-辣根过氧化物酶(HRP)复合物(HIVAg-HRP)，HIVAg-HRP喷样区接受待测HIV抗体(HIVAb)加样及洗涤液加样；工作电极处所在的检测区接受底物TMB溶液。其中HIV抗体加样需要自行加样，之后洗涤液和底物TMB溶液需要按序加样，中间间隔十数分钟。洗涤液能够通 过层析作用充分洗脱检测区处未与HIV抗原(S-HIVAg)结合的其他物质，使其移动出检测区，在检测区保留下“Ag-Ab-Ag(S-HIVAg—HIVAb—HIVAg-HRP)”型夹心结合物；之后加入的底物TMB和HRP发生酶催化反应，从而输出电流信号。
以具有微流控结构的检测芯片为例，部分具有微流控结构的检测芯片的使用场景中需要多次按序加样，例如用于农药检测的微流控电化学生物芯片和用于免疫检测的微流控电化学生物芯片。以艾滋病抗体(HIVAb)免疫检测用微流控电化学生物芯片为例，其工作电极上固定了HIV抗原(S-HIVAg)，为检测区。其中HIV抗体(HIVAb)需要自行加样，之后，一次洗涤液、HIV抗原-酶复合物(HIVAg-HRP)、二次洗涤液和TMB底物溶液需要按序加样，各自中间间隔十数分钟。一次洗涤液通过动力微流充分洗涤检测区未与HIV抗原(S-HIVAg)结合的其他物质，使其移动出检测区；随后，HIV抗原-酶复合物通过动力微流进入检测区，与检测区被捕获的HIV抗体结合，在检测区形成“Ag-Ab-Ag(S-HIVAg—HIVAb—HIVAg-HRP)”型夹心复合物；二次洗涤液通过动力微流充分洗脱未被捕获的HIV抗原-酶复合物；之后加入的TMB底物和HRP发生酶催化反应，从而输出电流信号。
附图说明
图1为本发明的实施例一内部安装结构示意图。
图2为图1的A局部放大图。
图3为本发明的实施例一使用状态变化图。
图4为本发明的实施例二使用状态变化图。
图5为图4的B局部放大图。
图6为图4的C局部放大图。
图7为本发明的实施例四中接液管结构示意图。
图8为本发明的实施例一配合用的层析式电化学生物芯片结构图。
图9为本发明的实施例一配合层析式电化学生物芯片进行测试的结果图。
图中：1.加样管；2.第一出液口；3.第二出液口；4.溶液段；5.活塞；6.活塞杆；7.第一出液管；8.连通管；9.第二出液管；10.滞留腔；51.槽；81.第一连通口；82第二连通口；A.芯片前端；B.芯片后端；11.工作电极；21.样品垫；22.层析膜；23.吸水垫；24.HIVAg-HRP喷样区；25.检测区；31.下盖；32.上盖；321.用于底物释放的加样孔；322.用于HIV抗体加样及洗涤液释放的加样孔；33.前端开口。
具体实施方式
下面结合附图对本发明做进一步的说明。
参阅图1-9，本说明书所附图式所绘示的结构、比例、大小等，均仅用以配合说明书 所揭示的内容，以供熟悉此技术的人士了解与阅读，并非用以限定本发明可实施的限定条件，故不具技术上的实质意义，任何结构的修饰、比例关系的改变或大小的调整，在不影响本发明所能产生的功效及所能达成的目的下，均应仍落在本发明所揭示的技术内容得能涵盖的范围内。同时，本说明书中所引用的如“上”、“下”、“左”、“右”、“中间”及“一”等的用语，亦仅为便于叙述的明了，而非用以限定本发明可实施的范围，其相对关系的改变或调整，在无实质变更技术内容下，当亦视为本发明可实施的范畴。
实施例一：
一种按序加样装置，为串行模式，参考图1，加样管1为一根整管，包括管壁，加样管1内一端向着另一端设置有四个存储空间，任意一个存储空间均包括预装于管内的溶液段4及其后部的活塞5；所述管壁上设置有可与其内溶液相通的第一出液口2、三个第二出液口3；所述第一出液口2由堵头预封闭(堵头未在图中示出)；三个第二出液口3分别设置在其前一个存储空间的活塞5封闭的区域内，所述活塞5起区隔、出液口的开关及推动作用。第四个活塞的后端敞开，可以用于接触动力装置。
向加样管中预装液体溶液的方法是在管壁上部开孔，从孔中注入溶液，而后将该开孔封闭(图中未示出)。
除第一出液口预封闭外，参考图2，管壁上的第二出液口下端向外敞开，但在初始时间，其上端在活塞的覆盖范围内，因此任一第二出液口均未能和后一个存储空间的溶液段连通。
图3显示串行按序加样装置的使用过程。开始使用时，第四存储空间的活塞5敞开于外部的一端与活塞杆6接触，活塞杆6可以被手动推动，或被步进电机驱动(步进电机图上未示出)。当需要进行按序加样时，开放预封闭的所述第一出液口2，通过活塞杆6推动接触的活塞5，管内溶液及活塞整体向前移动，第一存储空间的溶液段4从第一出液口2释出，第一存储空间的活塞5由于向前移动将第二出液口3暴露至第二存储空间的溶液段4中。此时推动停止，由于大气压的作用，虽然第二出液口3已与第二存储空间的溶液段4连通，该溶液也不会释出。待推动重新进行时，该溶液段的溶液才从第二出液口3中释出。如此操作，前一个存储空间的活塞5运动至使后一溶液段4与第二出液口3连通的位置，使得后一溶液段4被推动时可经由第二出液口3释出，各溶液段的溶液可以按序释出，从而达到按序加样的效果。若采用步进电机进行驱动，步进电机的出口与滚珠丝杠连接，滚珠丝杠上设置有丝杠螺母，丝杠螺母的端面与活塞杆接触；步进电机与控制器连接，控制器通过延时控制丝杠螺母间歇推动活塞杆运动，通过对控制器编程，可以对推动进程进行精确定位控制，也可以精确控制不同溶液加样的间隔时间。
参考图1至图3，出液口外可以设有螺纹或卡扣(图中未示出)，便于与出液管紧密连接。
本实施方案展示了一种包含四个存储空间的按序加样装置，根据实际需要，存储空间可以在两个至更多个之间变化。
由于在加样管中预装二个及以上的液体溶液，根据对活塞杆推动的进程控制和时间间隔控制，使用时，通过驱动器推动活塞杆运动，能根据按序加样的要求分别从加样口释出液体溶液，能够实现精确的按序加样，大幅提升了反应程序的简便程度，适合于制作便携式器件，更加适用于现场检测应用场景。
两个及以上的液体溶液可以是相同的，也可以是不同的。特别是当溶液段中溶液不同的情况下，不同的出液口单独接管，既可以避免溶液间的污染，也可以将不同的溶液引向不同的地方，避免附加其他复杂管路，仅通过活塞推动即可实现自动化按序在不同区域加样。
实施例二：
一种按序加样装置，为并排模式，参考图4、图5、图6，加样管1包括五个并排布置的独立腔体，五个存储空间分别位于五个独立腔体内，每个独立腔体包括管壁及其内的溶液段4和活塞5，本实施方案采用并排布置的独立腔体，整体长度更加合理，避免了串行模式下存在较多存储空间时活塞杆伸出加样管长度过长的技术问题。具体地，第一独立腔体的管壁上设置有连通溶液段4的第一出液口2，其他四个独立腔体的管壁上均设置有连通溶液段4前端的第二连通口82；第一至第四独立腔体还包含覆盖在活塞5之下的且靠近溶液段4末端的第一连通口81及其临近设置的第二出液口3，第二出液口3和第一连通口81初始状态不连通；第二至第五独立腔体管壁上的第二连通口82通过连通管8与前一独立腔体的第一连通口81连通；第一至第四独立腔体的活塞5在远离溶液段4的一端外圈设置有沿着轴向布置的槽51，所述的槽51的长度能覆盖第一连通口81与第二出液口3。第一出液口2初始状态时预封闭，以便使用时解除封闭。
任意一个所述独立腔体内的活塞5敞开于外部的一端均用于与分别设置的活塞杆6接触，每个活塞杆6可与对应的驱动器连接。
本实施例中，第一出液口2与第一出液管7连通，以便引导到所需加样的区域。可采用阀门(图中未示出)接续第一出液管7并实现预封闭。
图4显示并排按序加样装置的使用过程。当需要按序加样时，旋转阀门，第一出液口2与第一出液管7连通，第一独立腔体的活塞杆6被驱动，推动活塞5，将其前端的溶液段4经由第一出液口2和第一出液管7向外释出，当溶液释出完成时，活塞5上的槽51运动至覆盖第一连通口81与第二出液口3的位置，使得第一连通口81与第二出液口3通过槽51相连 通，从而也经由连通管8与后一独立腔体的溶液段4相连通；后一个独立腔体中的活塞5被其对应的活塞杆6推动时，其对应的溶液段4中的溶液顺序通过后一个独立腔体的第二连通口82、连通管8、前一个存储空间的第一连通口81、槽51、第二出液口3释出。第二出液口3接续相应的第二出液管9，以便引导到所需加样的区域。依次操作，可按序将五个独立腔体的液体释出。
实施例三：
在实施例一或实施例二的基础上，一种按序自动加样系统，包括按序加样装置，活塞杆6，驱动器，所述的驱动器包括步进电机，步进电机的出口与滚珠丝杠连接，滚珠丝杠上设置有丝杠螺母，丝杠螺母的端面与活塞杆6接触；步进电机与控制器连接，控制器通过延时控制丝杠螺母间歇推动活塞杆6运动。本发明通过控制器控制步进电机的运动，控制器控制内容包括推进启动的时间间隔、推进速度，从而使反应所需加入的溶液按设定顺序和时间间隔自动加入到设定区域，大幅提升了反应自动化程度，通过活塞杆使不同部位的液体在设定的时间进行释放，排除了人工操作误差，大幅提升了反应结果的精确度。所述丝杠螺母可以与活塞杆6连接固定，避免活塞杆误移动而驱动活塞。
实施例四：
参考图7，第一出液口2和第二出液口3可以分别连接有接液管，接液管中部设置有容纳溶液的滞留腔12，接液管的出口与相应的溶液接纳端入口对应布置。
实施例五：
通过按序加样装置实现层析式电化学双抗原夹心法检测艾滋病毒(HIV)抗体的实验：
实验试剂：
HIV抗原(HIVAg)(广东菲鹏生物有限公司)；辣根过氧化物酶(HRP)标记的HIV抗原(HIVAg-HRP)(广东菲鹏生物有限公司)；HIV-I型抗体(HIV-IAb)(5k)(广东菲鹏生物有限公司)；牛血清白蛋白(BSA)(生工生物工程上海股份有限公司)；酪蛋白(casein)(sigma公司)；3,3′,5,5′-四甲基联苯胺(TMB)底物溶液(0.4g/L)(Thermo公司)；1*PBS缓冲溶液；1*PBST洗涤液(1L的1*PBS缓冲液中添加了500μL Tween-20)；pH 9.6碳酸钠缓冲溶液(包被液)。
实验原材料：
丝网印刷电极芯片(浙江纳智汇生物科技有限公司)、玻璃纤维素膜(上海杰一生物技术有限公司，产品型号：GL0145)、吸水垫(上海杰一生物技术有限公司，产品型号：H5015)、硝酸纤维素膜(沃特曼公司，WhatmanAE99)；
实验用的检测结构：
参考图8，实验检测结构包括层析式电化学检测芯片，其中A为芯片前端；B为芯片后端，层析式电化学检测芯片的上方设置有上盖32，层析式电化学检测芯片的下方设置有下盖31,上盖32和下盖31的对合面上均设置有长方形的开口槽，电化学检测芯片安装在开口槽内，开口槽的开口均为前端开口33，上盖32的顶面从前到后依次设置有用于HIV I抗体加样及洗涤液释放的加样孔322、用于底物释放的加样孔321；层析式电化学检测芯片由电极芯片及紧密粘贴在其上的层析试纸条组成，其中电极芯片包括工作电极11以及分别位于该工作电极11两侧的对电极以及参比电极，三者通过丝网印刷工艺制作于基板上形成电极芯片，三个电极的引线集合于芯片的前端，引线从试纸条的前端伸出。工作电极11上固定有HIV抗原(S-HIVAg)；其中层析试纸条由样品垫21、层析膜22、吸水垫23依次叠合而成，样品垫21靠近A端位置设置有HIVAg-HRP喷样区24，检测区25位于样品垫21的中部，工作电极11位于检测区25的范围内；
实验过程：
1)HIV抗原(HIVAg)用pH 9.6的包被液稀释到100μg.mL-1，取6μL滴加在电极芯片的工作电极11上，4℃过夜孵育；
2)将工作电极11表面用1*PBS冲洗以除去包被液，N2吹干，之后在电极芯片的三电极区域滴加1％BSA+1％Casein混合封闭液(1*PBS配置)50μL，37℃封闭2h。用1*PBS冲洗以去除封闭液，吹干，备用。此时工作电极上的HIV抗原为包被抗原，简称为S-HIVAg；
3)将HIVAg-HRP用上述封闭液稀释到1：200倍，用商业用喷膜仪在玻璃纤维素膜样品垫21的一端喷样，形成喷样区，于25℃的恒温恒湿箱中干燥2h，备用；
在步骤2)的电极芯片上粘贴固定了HIVAg-HRP的玻璃纤维素膜样品垫21，层析膜22，吸水垫23三种材料，三种材料之间接续处有叠合，最终形成层析式电化学检测芯片。位置关系上，HIVAg-HRP喷样区24在靠近A端的位置，用于HIV-I抗体加样及洗涤液释放的加样孔33在HIVAg-HRP喷样区24正上方；样品垫21覆盖工作电极11，对应覆盖处为检测区25，层析膜不覆盖电极。
1*PBST洗涤液及HRP酶催化底物TMB溶液被预封装在类似于实施例一中的串行按序加样装置的加样管1中；参考图1，第一存储空间包含的溶液段4为150μL的1*PBST洗涤液，之后接续有第一段活塞5，第二存储空间包含的溶液段4为100μL的底物TMB溶液，之后接续有第二段活塞5，第二段活塞5末端与推动用的活塞杆6接触，推动活塞杆运动的丝杠由可编程控制的步进电机驱动。加样管1的第一出液口与芯片的加样孔322对应；第二出液口与芯片的加样孔321对应。
效价为5k的HIV-I抗体用上述封闭液按不同倍数稀释(1：1000、1：500、1：100、1： 20)，取150μL滴加在加样孔322处，形成HIV-I抗体与HIVAg-HRP免疫反应的结合物，并层析移动到检测区，与检测区的HIV抗原(S-HIVAg)形成“Ag-Ab-Ag(S-HIVAg—HIV-I Ab—HIVAg-HRP)”型夹心结合物；
上述加样15min后，解除加样管的预封闭，由程序控制步进电机工作，通过推动活塞杆驱动活塞5，使得封装的1*PBST洗涤液通过第一出液口2释出，经由加样孔322释放到层析式电化学检测芯片的试纸条上，在洗涤液释放完成的同时，第二出液孔露出，推动停止；10min后，由程序控制重新推动活塞杆，驱动活塞，使封装的底物TMB溶液从第二出液孔释出，经由加样孔321释放到层析式电化学检测芯片的工作电极11所对应的试纸条检测区25上。
上述10min的间隔是为了洗涤液能够通过层析作用充分洗脱检测区处未与HIV抗原(S-HIVAg)结合的其他物质，使其移动出检测区；之后加入的底物TMB和HRP发生酶催化反应，通过时间电流曲线(i-t)采集电流数据；测试电位为-0.1V，测试时间50s后的电流信号值为输出检测数据。
不含HIV的封闭液的测试数据为空白背景数据。
结果分析：HIV-I抗体稀释1：1000、1：500、1：100、1：20，检测结果参考图9，即使抗体经1：1000倍稀释，也可以和背景信号明显区分。
具体实施时，可灵活选择本发明的串排模式按序加液结构与并排模式按序加液结构来进行按序加液，可使便携式器件适应多种应用场景。
以上仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。

Claims (10)

  1. 一种按序加样装置，包括加样管(1)，所述加样管(1)包括管壁，其特征在于，
    所述加样管(1)内一端向着另一端设置有N个存储空间，任意一个存储空间均包括预装于管内的溶液段(4)及其后部的活塞(5)，N大于1；
    所述管壁上设置有可与第一存储空间内溶液段(4)相通的第一出液口(2)、N-1个第二出液口(3)；所述第一出液口(2)预封闭；所述N-1个第二出液口(3)分别设置在其前一个存储空间活塞(5)封闭的区域内，所述活塞(5)起区隔、出液口的开关及推动作用；
    所述活塞(5)敞开于外部的一端用于接触动力装置，当需要进行按序加样时，开放预封闭的所述第一出液口(2)，通过动力装置推动对应接触的活塞(5)，第一存储空间的溶液段(4)的溶液释出，前一个存储空间的活塞(5)运动至使后一个存储空间的溶液段(4)与第二出液口(3)连通的位置，使得后一个存储空间溶液段(4)被推动时可经由第二出液口(3)释出；
    所述的第一出液口(2)预封闭是指第一存储空间的溶液段(4)不能经由第一出液口(2)与外界连通。
  2. 根据权利要求1所述的一种按序加样装置，其特征在于，所述的动力装置包括活塞杆(6)，活塞杆(6)与能做直线运动的驱动器连接，驱动器能推动活塞杆(6)带动活塞(5)向着其对应的溶液段(4)方向运动。
  3. 根据权利要求1所述的一种按序加样装置，其特征在于，所述加样管(1)为一根整管，整管内一端向着另一端依次设置有N个连续的存储空间，所述第一出液口(2)和所述的N-1个第二出液口(3)分别间距布置；所有第二出液口(3)均分别位于前一个存储空间的活塞(5)下方；最末端的活塞(5)敞开于外部的一端用于接触动力装置。
  4. 根据权利要求1所述的一种按序加样装置，其特征在于，
    所述加样管(1)包括N个并排布置的独立腔体，N个存储空间分别位于N个独立腔体内；
    每个独立腔体包括管壁及其内的溶液段(4)和活塞(5)，第一独立腔体的管壁上设置有连通溶液段(4)的第一出液口(2)，其他N-1个独立腔体的管壁上均设置有连通溶液段(4)的第二连通口(82)，第一出液口(2)预封闭；
    每个独立腔体还包含覆盖在活塞(5)之下的且靠近溶液段(4)一端的第一连通口(81)及其临近设置的第二出液口(3)，第二出液口(3)和第一连通口(81)初始状态不连通；N-1个独立腔体管壁上的第二连通口(82)通过连通管(8)与前一独立腔体的第一连通口(81)连通；
    每一个独立腔体的活塞(5)在远离溶液段(4)的一端外圈设置有沿着轴向布置的槽(51)， 所述的槽(51)的长度能覆盖第一连通口(81)与第二出液口(3)；当活塞(5)被推动时，所述的槽(51)随之移动，当槽(51)达到同时覆盖第一连通口(81)与第二出液口(3)的位置时，第一连通口(81)与第二出液口(3)通过槽(51)相连通，使得第二出液口(3)与其后一个独立腔体的溶液段(4)连通。
  5. 据权利要求4所述的一种按序加样装置，其特征在于，活塞(5)敞开于外部的一端均用于与动力装置接触，所述动力装置分别驱动。
  6. 根据权利要求1-5任一项所述的一种按序加样装置，其特征在于，所述的第一出液口(2)和所述的N-1个第二出液口(3)分别连接有出液管。
  7. 根据权利要求6所述的一种按序加样装置，其特征在于，出液管中部设置有容纳溶液的滞留腔，出液管的出口与相应的溶液接纳端入口对应布置。
  8. 一种按序自动加样系统，包括权利要求1-5任一项所述的按序加样装置，活塞杆(6)，驱动器，其特征在于，
    所述驱动器包括步进电机，步进电机的出口与滚珠丝杠连接，滚珠丝杠上设置有丝杠螺母，丝杠螺母的端面与活塞杆(6)接触；步进电机与控制器连接，控制器通过延时控制丝杠螺母间歇推动活塞杆(6)运动。
  9. 一种按序加样自动检测系统，包括权利要求1-5任一项所述的按序加样装置，动力装置，检测装置，其特征在于，所述按序加样装置的第一出液口(2)和所述的N-1个第二出液口(3)分别对接到检测装置的不同加样入口，所述动力装置自动驱动所述按序加样装置的活塞使得液体按序释出。
  10. 根据权利要求9所述的一种按序加样自动检测系统，其特征在于，所述检测装置为层析式结构或微流控结构的检测芯片。
PCT/CN2023/085398 2022-04-19 2023-03-31 一种按序加样装置及自动加样系统 WO2023202351A1 (zh)

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JP2009210354A (ja) * 2008-03-03 2009-09-17 Yaskawa Electric Corp 液体試料分注装置および駆動方法
DE102009023068A1 (de) * 2009-05-28 2010-12-02 RATIONAL Technische Lösungen GmbH Dosiergerät insbesondere für analytische Aufgaben
CN102202791A (zh) * 2008-11-05 2011-09-28 生物梅里埃公司 用于制备和/或处理生物样品的装置
WO2013046417A1 (ja) * 2011-09-30 2013-04-04 ミライアル株式会社 マイクロ流路チップ
CN103037971A (zh) * 2010-04-08 2013-04-10 阿瓦克塔有限公司 用于将液体试剂顺序地配送到反应腔室的设备
CN113711051A (zh) * 2019-04-16 2021-11-26 结盟遗传学有限公司 分析物收集装置、利用其的分析物收集方法及分析物检查系统

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2009210354A (ja) * 2008-03-03 2009-09-17 Yaskawa Electric Corp 液体試料分注装置および駆動方法
CN102202791A (zh) * 2008-11-05 2011-09-28 生物梅里埃公司 用于制备和/或处理生物样品的装置
DE102009023068A1 (de) * 2009-05-28 2010-12-02 RATIONAL Technische Lösungen GmbH Dosiergerät insbesondere für analytische Aufgaben
CN103037971A (zh) * 2010-04-08 2013-04-10 阿瓦克塔有限公司 用于将液体试剂顺序地配送到反应腔室的设备
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