A pipeline scheduling method for sample pretreatment using magnetic beads

Abstract

The invention provides a pipeline scheduling method for magnetic bead method sample pretreatment, which divides the mixing time and magnetic absorption time in the sample magnetic bead method pretreatment process into a plurality of mixing cycles and magnetic absorption cycles, completes the mixing and magnetic absorption in sequence, and completes the injection in sequence according to the required number of injections. The method is completed in a specific magnetic bead method sample pretreatment instrument, each sample is placed in a reaction cup, and the scheduling is completed by moving each reaction cup between different scheduling hole positions through a mechanical gripper, and the injection is completed through a liquid injection mechanical arm. Single-tube sample loading and emergency queue jumping are supported, and the process parameters of the sample pretreatment can be freely configured. Samples with different pretreatment parameters can still be processed in the pipeline and samples are sequentially discharged in each reaction cycle, which does not affect the overall beat and efficiency, ensures the high efficiency of the magnetic bead method sample pretreatment, and smoothly connects with various automatic detection equipment.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a pipeline scheduling method for magnetic bead method sample pre-treatment. Background Art

[0002] Mass spectrometry is an ion charge-to-mass ratio analysis method that can be used to analyze isotopic composition, organic structure, and elemental composition. Mass spectrometry can provide rich structural information in one analysis. Combining separation technology with mass spectrometry is a breakthrough in separation science. Mass spectrometry technology is developing rapidly. With the development of mass spectrometry technology, the application field of mass spectrometry technology is becoming more and more extensive. Since the matrix of biological samples is complex and the target substances to be detected are mostly endogenous compounds, such as serum samples containing proteins, peptides, amino acids, lipids, sugars, inorganic salts, vitamins, organic acids, hormones, etc. Therefore, in order to reduce matrix interference, extend the service life of the chromatographic column, prevent system blockage and damage, and improve detection sensitivity and specificity, it is often necessary to perform pre-treatment such as precipitation, extraction, purification, and enrichment on the sample.

[0003] At present, the methods used in mass spectrometry pretreatment are basically: protein precipitation, liquid-liquid extraction, solid phase extraction, magnetic extraction, etc. Among them, protein precipitation, liquid-liquid extraction, and solid phase extraction are gradually withdrawn from the mainstream choice due to many factors such as cumbersome processing procedures, large differences in processing technology, long process time, many influencing factors, high requirements for the professional quality of operators, and high costs. On the contrary, magnetic bead extraction is increasingly recognized and selected by the market because of its many advantages such as simple process, convenient operation, good consistency, high degree of automation, and high efficiency.

[0004] Magnetic bead extraction is a method of separating and enriching target molecules from samples, and is widely used in the fields of biological science, medical research, and industrial production. It uses magnetic particles to specifically bind to target molecules, and achieves separation and concentration of target molecules with the assistance of a magnetic field, and can be used for sample pretreatment. Under the action of the magnetic field, the magnetic beads bound to the target molecules gather together, and then the target molecules are separated from the magnetic beads by elution, and the magnetic beads are adsorbed and removed by a magnetic rod, thereby achieving separation and concentration of the target molecules. The remaining sample after magnetic bead extraction can be used for subsequent mass spectrometry detection.

[0005] At present, most of the magnetic bead extraction methods on the market have adopted the relevant technologies of nucleic acid extraction, and continue to use 96-well plates as consumables. The liquid is pipetted one by one in each well, and then the effective substances are transferred by pipetting or magnetic bead transfer. The transfer of some wells still requires human intervention. Overall, although it has been greatly improved compared with traditional methods, it still has obvious shortcomings. Its main defects are described as follows: (1) Low efficiency. Using 96-well plates as consumables, it is necessary to wait until all wells have completed the same step before transferring to the next process, which takes a long time to wait; (2) Poor compatibility. Only a single extraction process can be implemented. For different test projects, even some R&D projects do not have the ability to implement it; (3) Poor versatility. The back-end can only be used for the sample injection device of the 96-well plate, and the adaptability is relatively single; (4) Single function. It cannot be connected to the existing laboratory detection pipeline and is isolated from the detection system; (5) The degree of automation is not enough. Many of them are only semi-automated, and some actions still require human intervention.

[0006] The most critical thing is to use a 96-well plate placed in a magnetic bead extractor to complete the relevant operations. The 96-well plate is pre-loaded with magnetic bead activators, magnetic bead eluents, magnetic bead washing solutions, etc. Each reagent needs to be pre-loaded and sealed in a row. That is to say, a 96-well plate is pre-loaded with all the reagents required for magnetic bead extraction of 16 samples. Therefore, when performing magnetic bead extraction operations, exactly 16 samples need to be processed each time; if there are 15 samples, the magnetic bead processing reagents (including magnetic bead liquid, magnetic bead activators, magnetic bead eluents, magnetic bead elution solutions, etc.) of one sample will be wasted, because after the well plate is placed in the magnetic bead extractor, regardless of whether the corresponding well position is loaded with samples, the magnetic rod corresponding to the well position will complete all magnetic bead extraction operations for it; if there are 17 samples, two 96-well plates are required, the first 96-well plate processes 16 samples, and the second 96-well plate processes 1 sample, and the magnetic bead processing reagents of 15 samples will be wasted in the second 96-well plate. Therefore, in reagent operation, a large amount of magnetic bead treatment reagents are often wasted, which increases the detection cost and the detection fee is difficult to calculate.

[0007] It can be seen that the magnetic bead sample pretreatment instrument using a 96-well plate cannot realize the single-tube sample loading operation, which brings great inconvenience to the magnetic bead extraction. To this end, the team of the present invention has developed a fully automatic device (CN118150739) that can realize magnetic bead processing and mass spectrometry detection in the early stage of research, which can realize the single-tube sample loading operation, and can be fully automatically docked with the high-throughput LC-MS / MS detection module to complete the fully automatic pipeline processing from sample pretreatment to detection. However, the fully automatic pipeline processing equipment still has problems in actual use. For example, the pretreatment process of each sample is required to be as consistent as possible, and the mixing time, magnetic absorption time, and injection times must be kept consistent. Once the pretreatment process of some samples is inconsistent, it is necessary to pause and reset the pretreatment process parameters and restart the equipment, otherwise it will cause the intermediate waiting time to be too long, making it difficult to sample the pipeline, and it cannot be docked with the subsequent mass spectrometry detection pipeline. It is isolated from the detection system and even requires human intervention to solve it, and can only achieve semi-automation. In addition, the device can only be used for solid-phase magnetic beads combined with magnetic mesoporous composite materials and microstructured domains. The main reason is that compared with immunomagnetic beads, the sample pretreatment process of solid-phase magnetic beads is more unified and the pretreatment process parameters rarely need to be adjusted.

[0008] It can be seen that the pre-treatment parameters of the existing magnetic bead method pre-treatment instruments are basically fixed, or the parameters need to be adjusted at the expense of efficiency. The overall adjustability is poor and cannot adapt to the current use environment that requires multi-process and multi-process compatibility; it is also impossible to achieve compatibility when developing new methodologies. Therefore, it is urgent to find a magnetic bead method sample pre-treatment method with adjustable pre-treatment parameter configuration and scheduling functions, which can realize the sequential pre-treatment of samples with different magnetic bead pre-treatment parameters, and can still be sampled sequentially according to regulations, smoothly dock with the assembly line, adapt to various types of magnetic beads, and ensure the overall efficiency of magnetic bead method pre-treatment. Summary of the invention

[0009] In order to solve the above problems, the present invention provides a pipeline scheduling method for magnetic bead method sample pretreatment, which divides the mixing time and magnetic attraction time in the sample magnetic bead method pretreatment process into multiple mixing cycles and magnetic attraction cycles, and completes the mixing and magnetic attraction in sequence, and completes the injection in sequence according to the required number of injections. The method is completed in a specific magnetic bead method sample pretreatment instrument, and each sample is placed in a reaction cup respectively. The scheduling is completed by moving each reaction cup between different scheduling hole positions through a mechanical gripper, and the injection is completed by a liquid injection mechanical arm. Single-tube sampling and emergency queue jumping are supported, and the process parameters of sample pretreatment can be freely configured. Samples with different pretreatment parameters can still be processed in the pipeline and samples are taken out in sequence in each reaction cycle, which does not affect the overall beat and efficiency, ensures the high efficiency of magnetic bead method sample pretreatment, and smoothly connects to various fully automatic detection equipment.

[0010] On the one hand, the present invention provides a pipeline scheduling method for magnetic bead sample pretreatment, wherein the sample pretreatment includes mixing and magnetic absorption; the mixing is divided into a plurality of mixing cycles according to the mixing time and is completed in sequence and progressively, and the mixing cycle is the minimum unit time that can be measured when the sample is mixed; the magnetic absorption is divided into a plurality of magnetic absorption cycles according to the magnetic absorption time and is completed in sequence and progressively, and the magnetic absorption cycle is the minimum unit time that can be measured when the sample is magnetically absorbed.

[0011] The sample pretreatment process of the magnetic bead method is mostly relatively uniform: inject sample into the reaction cup - inject magnetic bead solution into the reaction cup - mix 1 - stand still and absorb magnet 1 - extract waste liquid 1 - (may inject other reagent solution - mix - stand still and absorb magnet - extract waste liquid) - rinse 1 liquid injection - mix 2 - stand still and absorb magnet 2 - extract waste liquid 2 - rinse 2 - mix 3 - stand still and absorb magnet 3 - extract waste liquid 3 - eluent injection - mix 4 - stand still and absorb magnet 4 - extract sample to another reaction cup for detection. It can be seen that the core parameters of the whole process are: the number of reagent injections, mixing time and magnetic absorption time.

[0012] Existing magnetic bead sample pretreatment instruments are usually simple serial single-line modes, that is, each step is completed separately for each series of samples. For example, series A samples need to be mixed for 1 minute and magnetically attracted for 3 minutes, and series B samples need to be mixed for 3 minutes and magnetically attracted for 2 minutes. It is necessary to complete all series A samples first, adjust the parameter settings, and load series B samples.

[0013] In order to be able to connect with the subsequent detection module for a fully automatic assembly line and try not to affect the overall rhythm and efficiency, the pre-treatment instrument needs to try to discharge the sample into the detection module on time in each reaction cycle (for example, 1 minute) to ensure the on-line sampling and detection. Therefore, the existing magnetic bead method sample pre-treatment instrument is only suitable for samples with consistent pre-treatment parameters to maintain on-line sampling. Although it can also be used for emergency sampling, it is necessary to ensure that the pre-treatment parameters for emergency sampling are consistent with other sample parameters; once it is necessary to process samples with differences in mixing time, magnetic absorption time, injection times, etc., the overall rhythm of the assembly line will be disrupted, and it will no longer be possible to ensure that the sample is discharged on time in each reaction cycle.

[0014] The pipeline scheduling method based on magnetic bead method sample pretreatment provided by the present invention divides the mixing time and magnetic attraction time of each sample into multiple mixing cycles and magnetic attraction cycles, so that subsequent samples can be sorted in sequence and gradually follow up, and the mixing and magnetic attraction are completed one by one in sequence according to the rhythm of the mixing cycle or the magnetic attraction cycle. Even if the mixing time or magnetic attraction time of each sample is different, it can be decomposed into multiple mixing cycles and magnetic attraction cycles, so that subsequent samples can still follow up in sequence, thereby greatly eliminating the negative impact of different mixing times or magnetic attraction times on pipeline processing.

[0015] Furthermore, the sample pretreatment also includes liquid injection; the liquid injection is completed in sequence according to the required number of injections.

[0016] Furthermore, the method completes sample pretreatment by means of a magnetic bead method sample pretreatment instrument, and the magnetic bead method sample pretreatment instrument is provided with a scheduling module, and the scheduling module includes a liquid injection scheduling module, a mixing scheduling module and a magnetic attraction scheduling module.

[0017] In order to realize the free configuration of process parameters such as the number of injections, mixing time, and magnetic absorption time in the magnetic bead method sample pretreatment without affecting the overall beat and efficiency, an injection scheduling module, a mixing scheduling module, and a magnetic absorption scheduling module are set on the magnetic bead method sample pretreatment instrument, and realized through intelligent scheduling.

[0018] Furthermore, the method comprises the following steps:

[0019] (1) Set the mixing cycle and magnetic absorption cycle of the sample;

[0020] (2) According to the mixing time and magnetic absorption time required for the sample pre-treatment by magnetic beads, determine the number of mixing cycles and magnetic absorption cycles required for mixing and magnetic absorption respectively;

[0021] (3) According to the number of injections, mixing cycles, and magnetic absorption cycles required for the sample to be pre-treated by the magnetic bead method, the number of scheduling holes required for the sample in the injection scheduling module, mixing scheduling module, and magnetic absorption scheduling module should be determined;

[0022] (4) The samples are processed in the scheduling wells within the number of scheduling wells in sequence.

[0023] In some methods, the mixing period and the magnetic attraction period are both 1 minute; if the mixing time of the series a samples is 2 minutes, the mixing needs to be completed in two mixing scheduling holes; if the magnetic attraction time of the series b samples is 3 minutes, the magnetic attraction needs to be completed in three magnetic attraction scheduling holes; if the series a samples need to be injected twice and the series b samples need to be injected three times, the series a samples need to be injected in two injection scheduling holes and the series b samples need to be injected in three injection scheduling holes.

[0024] Furthermore, the sample pretreatment steps include: liquid injection - mixing - magnetic absorption - waste liquid absorption; the number of liquid injections includes one or more times, and each time the liquid is injected, mixing - magnetic absorption - waste liquid absorption is required.

[0025] In some methods, each scheduling hole in the injection scheduling module represents one injection. For example, the injection scheduling module is provided with A1, A2, A3, A4, A5, A6, and A7. The a series reaction cups (10 pieces) need to be injected twice, and the b series reaction cups (10 pieces) need to be injected three times. Then, after each reaction cup of the a series completes the first injection from A1, it enters the subsequent mixing, magnetic absorption, and waste liquid absorption, and then returns to A2 for the second injection. After each reaction cup of the b series completes the first injection from A1, it enters the subsequent mixing, magnetic absorption, and waste liquid absorption, and then returns to A2 for the second injection, enters the subsequent mixing, magnetic absorption, and waste liquid absorption, and then returns to A3 to complete the third injection, enters the subsequent mixing, magnetic absorption, and waste liquid absorption, and then enters the elution step. Samples that require different numbers of injections can be scheduled and arranged in this module at the same time. Each time an injection is completed, it will move forward to a scheduling hole position, and the sample injection will be completed in sequence in the assembly line, and then enter the subsequent mixing step. The different injection times will not affect the sample's automated pre-processing rhythm. When switching from series a samples to series b samples, only the first series b sample needs to increase the injection time, which adds a round of injection, mixing, magnetic absorption, and waste liquid absorption, resulting in a delay in the sample output time. Starting from the second series b sample, the original assembly line automated pre-processing rhythm can still be maintained, and the original sample output speed can be maintained. If there is no scheduling hole position, starting from the series b samples, each sample needs to add a round of injection, mixing, magnetic absorption, and waste liquid absorption, resulting in a delay in the sample output time of each sample.

[0026] In some methods, each scheduling hole in the mixing scheduling module represents a mixing cycle (for example, 1 minute). For example, the mixing scheduling module is provided with B1, B2, B3, B4, B5, B6, and B7. The reaction cups of the a series need to be mixed for 2 minutes, and the reaction cups of the b series need to be mixed for 3 minutes. Then, each reaction cup of the a series is first mixed for 1 minute in B1, and then moved to B2 for 1 minute, and then enters the subsequent magnetic absorption and waste liquid absorption; each reaction cup of the b series is first mixed for 1 minute in B1, moved to B2 for 1 minute, moved to B3 for 1 minute, and then entered the subsequent magnetic absorption and waste liquid absorption, and then entered the elution step. Samples that require different mixing times can be scheduled and arranged in the module at the same time. After each mixing cycle is completed, one scheduling hole is moved forward, and the sample mixing is completed in sequence in the pipeline, and then the subsequent mixing steps are entered. The automated pre-processing rhythm of the sample will not be affected by the different mixing times. When switching from series a samples to series b samples, only the first series b sample needs to add 1 minute of mixing time, resulting in a 1 minute delay in sample output. Starting from the second series b sample, the original pre-processing rhythm of the assembly line can still be maintained, and the original sample output speed of 1 minute can be maintained. If there is no scheduling well position, starting from the series b samples, each sample needs to be mixed in the original well position for 3 minutes, which is 1 minute later than before, resulting in a 1 minute delay in the output of each sample in the series b samples.

[0027] In some methods, each scheduling hole in the magnetic attraction scheduling module can be magnetized, and each hole represents a magnetic attraction cycle (for example, 1 minute). For example, the magnetic attraction scheduling module is provided with C1, C2, C3, C4, C5, C6, and C7. The reaction cup of series a needs to be magnetized for 2 minutes, and the reaction cup of series b needs to be magnetized for 3 minutes. Then, each reaction cup of series a first magnetizes at C1 for 1 minute, then moves to C2 for 1 minute, and then enters the subsequent waste liquid absorption; each reaction cup of series b first magnetizes at C1 for 1 minute, moves to C2 for 1 minute, moves to C3 for 1 minute, and then enters the subsequent waste liquid absorption. Samples that require different magnetic absorption times can be scheduled and arranged in this module at the same time. After each magnetic absorption cycle is completed, one scheduling hole is advanced, and the sample magnetization is completed in sequence in the assembly line, and then the subsequent waste liquid absorption step is entered. The automated pre-processing rhythm of the sample will not be affected by the different magnetic absorption times. When switching from series a samples to series b samples, only the first series b sample needs to add 1 minute of magnetization time, resulting in a 1 minute delay in sample delivery. Starting from the second series b sample, the original pre-processing rhythm of the assembly line can still be maintained, and the original sample delivery speed of 1 minute can be maintained. If there is no scheduling hole, starting from the series b samples, each sample needs to be magnetized in the magnetization module for 3 minutes, which is 1 minute later than before, resulting in a 1 minute delay in sample delivery for each series b sample.

[0028] Furthermore, the sample is placed in a reaction cup; the reaction cup can be moved between different scheduling holes by a mechanical gripper until the pre-treatment of the sample is completed.

[0029] Furthermore, one scheduling hole position of the liquid injection scheduling module described in step (3) corresponds to one liquid injection into the reaction cup therein; one scheduling hole position of the mixing scheduling module corresponds to one reaction cup therein completing mixing for one reaction cycle time; one scheduling hole position of the magnetic attraction scheduling module corresponds to one reaction cup therein completing magnetic attraction for one reaction cycle time; after each reaction cup is processed at the scheduling hole position, it is removed for the next step of processing.

[0030] Furthermore, the injection in step (3) is completed by an injection robot, and the time required to complete one injection does not exceed the time of the mixing cycle or the magnetic attraction cycle.

[0031] An injection process includes the pipette needle sucking up the reagent, injecting it into the reaction cup and cleaning the pipette needle. If the time to complete an injection exceeds the time of a mixing cycle or a magnetic attraction cycle, the sample after each mixing or magnetic attraction cycle will need to wait for the injection to be completed before entering the next processing step, which will inevitably affect the overall rhythm of the assembly line operation. Therefore, the time required for an injection should not exceed the time of the mixing cycle or the magnetic attraction cycle.

[0032] In some embodiments, the liquid injection robot arm takes a total of 15 seconds to perform one liquid injection. When a mixing cycle and a magnetic attraction cycle are 1 minute, four liquid injections can be completed within this cycle.

[0033] In some embodiments, the liquid injection scheduling module, the mixing scheduling module and the magnetic attraction scheduling module are each provided with 7 scheduling holes. Then, the time required to complete the liquid injection of the 7 scheduling holes should not exceed the time of the mixing cycle or the magnetic attraction cycle (1 min). Therefore, two liquid injection robotic arms are required, and a maximum of 8 reaction cups can be filled in one cycle.

[0034] Furthermore, the samples include a series of samples, the a series of samples include samples a1, a2~an, the number of injections, mixing time and magnetic absorption time required for the pre-treatment reaction of the samples a1, a2~an are consistent; the scheduling hole positions of the injection scheduling module include A1, A2~An, the scheduling hole positions of the mixing scheduling module include B1, B2~Bx, and the scheduling hole positions of the magnetic absorption scheduling module include C1, C2~Cy, and n, x, and y are integers greater than or equal to 2, n=the number of injections required for the sample; x=the number of mixing reaction cycles required for the sample; y=the number of magnetic absorption reaction cycles required for the sample; the method includes the following steps:

[0035] Ⅰ. Load sample a1 into reaction cup a1 and place it at position A1 of the injection scheduling module to complete the injection;

[0036] II. Move the reaction cup a1 from A1 to B1 of the mixing scheduling module to perform the mixing of the first mixing cycle; and move it from B1 to Bx in sequence to complete the mixing of each mixing cycle; when a reaction cup is moved out of the A1, B1~Bx positions, immediately add the next reaction cup until the mixing of the reaction cup an is completed;

[0037] III. Move the reaction cup a1 from the Bx position to the C1 position of the magnetic attraction scheduling module to carry out the first magnetic attraction cycle; and move from C1 to Cy in sequence to complete the magnetic attraction of each magnetic attraction cycle; when a reaction cup is moved out of the A1, B1~Bx, C1~Cy positions, immediately add the next reaction cup until the magnetic attraction of the reaction cup an is completed;

[0038] V. After completing the magnetic attraction and absorbing the waste liquid, the reaction cup moves from the Cy position to the A2 position of the injection scheduling module to complete the second injection, and then completes the mixing and magnetic attraction in accordance with steps II and III, until n injections and corresponding mixing and magnetic attraction are completed.

[0039] In some methods, when the sample only needs one injection, one mixing cycle, and one magnetic attraction cycle, the pretreatment is the most basic and simplest processing flow, that is, reagent injection - mixing - magnetic attraction - waste liquid extraction - elution, the flow chart is shown in Figure 1 Existing magnetic bead pretreatment instruments also follow this process, but they must wait for a sample to complete the entire process of reagent injection - mixing - magnetic absorption - waste extraction - elution before they can rotate to the next sample for processing. The methodology is extremely fixed and rigid, and lacks adjustability and adaptability.

[0040] In some methods, if different projects have different requirements on the mixing time, for example, project A needs to be mixed for 1 cycle, project B needs to be mixed for 2 cycles, and project N needs to be mixed for n cycles, in order to adapt to this situation, the present invention can realize the requirements of different projects for different mixing cycles by designing the scheduling holes of the mixing scheduling module in a step-by-step progressive manner. The flow chart is shown as follows: Figure 2 As shown, the specific steps include:

[0041] (1) Move the injection scheduling module to the sample loading area and load the reaction cup into the A1 injection scheduling hole of the injection scheduling module;

[0042] (2) The injection robot transfers the solution into the reaction cup at the A1 injection scheduling hole;

[0043] (3) The gripper grabs the reaction cup that has completed the injection at the A1 injection scheduling hole and moves it to the B1 mixing scheduling hole for mixing;

[0044] (4) The gripper grabs the reaction cup that has completed the mixing cycle at the B1 mixing scheduling hole and moves it to the B2 mixing scheduling hole for mixing;

[0045] ………

[0046] (5) The gripper grabs the reaction cup that has completed the mixing cycle at the B(n-1) mixing scheduling hole and moves it to the Bn mixing scheduling hole for mixing;

[0047] (6) The gripper grabs the Bn mixing scheduling hole to complete the mixing cycle of the reaction cup and moves it to the C1 magnetic attraction scheduling hole for static magnetic attraction;

[0048] (7) The magnetic suction scheduling hole is moved to the waste liquid extraction area to complete the waste liquid extraction;

[0049] (8) Elution injection.

[0050] Figure 2 The solution shown only optimizes the design and compatibility of the mixing time parameter for different projects. However, in actual use, different projects have different requirements for magnetic attraction time.

[0051] In some ways, if different projects have different requirements for magnetization time, for example, project A requires 1 magnetization cycle, project B requires 2 magnetization cycles, and project N requires n magnetization cycles; in order to adapt to this situation, the present invention realizes the requirements of different projects for different magnetization cycles by designing the scheduling holes of the magnetization scheduling module in a step-by-step progressive manner. The flow chart is as follows: Figure 3 As shown, the specific steps include:

[0052] (1) Move the injection scheduling module to the sample loading area and load the reaction cup into the A1 injection scheduling hole of the injection scheduling module;

[0053] (2) The injection robot transfers the solution into the reaction cup at the A1 injection scheduling hole;

[0054] (3) The gripper grabs the reaction cup that has completed the injection at the A1 injection scheduling hole and moves it to the B1 mixing scheduling hole for mixing;

[0055] (4) The gripper grabs the reaction cup that has completed the mixing at the B1 mixing and scheduling hole and moves it to the C1 magnetic attraction scheduling hole for static magnetic attraction;

[0056] (5) The gripper grabs the reaction cup that has completed the magnetic attraction at the C1 magnetic attraction scheduling hole and moves it to the C2 magnetic attraction scheduling hole for static magnetic attraction;

[0057] ………

[0058] (6) The gripper grabs the reaction cup that has completed the magnetic absorption cycle at the C(n-1) magnetic absorption scheduling hole position and moves it to the Cn magnetic absorption scheduling hole position for magnetic absorption;

[0059] (7) The magnetic suction scheduling hole is moved to the waste liquid extraction area to complete the waste liquid extraction;

[0060] (8) Elution injection.

[0061] In some methods, the scheduling holes of the mixing scheduling module and the magnetic attraction scheduling module are designed in a step-by-step manner, which can effectively solve the problem of free setting of process parameters such as mixing time and magnetic attraction time for different projects. However, in the actual use of pretreatment, there is another very important parameter that needs to be freely configured in the method, that is, the actual number of types that need to be added; for example, project a only needs to add 1 reagent, and elution and injection can be carried out after completing the mixing-magnetic attraction-waste liquid extraction process. Project b needs to add 2 reagents, and elution and injection can be carried out after completing the mixing-magnetic attraction-waste liquid extraction process in sequence. Project n needs to add n reagents, and elution and injection can be carried out after completing the mixing-magnetic attraction-waste liquid extraction process in sequence.

[0062] In order to realize the free configuration of the last parameter, the present invention adds a liquid injection scheduling module on the basis of the mixing scheduling module and the magnetic attraction scheduling module, and its scheduling hole positions are also designed in a step-by-step manner. The flow chart is as follows Figure 4 As shown,

[0063] The specific steps can be simplified as follows:

[0064] (1) Move the injection scheduling module to the sample loading area and load the reaction cup into the A1 injection scheduling hole of the injection scheduling module;

[0065] (2) The injection robot transfers the solution into the reaction cup at the A1 injection scheduling hole;

[0066] (3) The gripper grabs the reaction cup that has completed the injection at the A1 injection scheduling hole and moves it to the B1 mixing scheduling hole for mixing; and moves from B1 to Bx in sequence to complete the mixing of each mixing cycle; when a reaction cup is moved out of the A1, B1~Bx positions, the next reaction cup is immediately added until the mixing of reaction cup an is completed;

[0067] (4) The gripper grabs the reaction cup and moves it from the Bx position to the C1 position of the magnetic attraction scheduling module to perform the first magnetic attraction cycle; and moves from C1 to Cy in sequence to complete the magnetic attraction of each magnetic attraction cycle; when a reaction cup is moved out of the A1, B1~Bx, C1~Cy positions, the next reaction cup is immediately added until the magnetic attraction of reaction cup an is completed;

[0068] (5) After completing the magnetic attraction and the waste liquid, the reaction cup is moved from the Cy position to the A2 position of the injection scheduling module to complete the second injection, and then the mixing and magnetic attraction are completed in accordance with steps (3) and (4) until n injections and the corresponding mixing and magnetic attraction are completed.

[0069] Furthermore, when the samples also include emergency treatment series samples and / or other series samples, the emergency treatment series samples can be inserted into the a series samples for emergency loading at any time. During the pretreatment process of the emergency treatment series samples and other series samples, they can have one or more of different injection times, different mixing times, or different magnetic absorption times compared with the a series samples.

[0070] In some embodiments, the reaction reagents injected by the injection scheduling module include one or more of a magnetic bead solution, a washing solution, and an eluting solution.

[0071] On the other hand, the present invention provides a method for enabling samples with different pre-treatment parameters to be processed in sequence in an assembly line and sampled out in sequence in each reaction cycle. The method adopts the method described above for processing, and the reaction cycle is the minimum unit time that can be measured when the sample is mixed and / or magnetically attracted.

[0072] The pretreatment method provided by the present invention realizes the caching of the mixing, magnetic attraction and liquid injection programs on demand and the intelligent step-by-step progressive scheduling through the intelligent system, thereby realizing the effect of free configuration of the process parameters of the whole process; at the same time, each workstation operates in parallel, and except for affecting the cycle time from the beginning to the end of the pretreatment of the first sample, the subsequent samples still have a frequency of one sample per reaction cycle, thereby ensuring the overall high efficiency of the pretreatment equipment.

[0073] On the other hand, the present invention provides a magnetic bead sample pretreatment instrument, in which the scheduling module divides each pretreatment process into multiple reaction cycles, and can select a matching number of scheduling wells according to the steps and time required for sample pretreatment in the reaction cup. After the reaction cup completes a reaction cycle in the well, it will be automatically moved forward to the adjacent scheduling well for the next reaction cycle until all reaction cycles are completed.

[0074] The existing magnetic bead sample pretreatment instrument is only suitable for samples with consistent pretreatment parameters to maintain the sample loading on the assembly line. The magnetic bead sample pretreatment instrument provided by the present invention, through the design and arrangement of the structure, adds a multi-station cache and intelligent scheduling module for the reaction tube, which can make the mixing time and magnetic absorption time of the sample be evenly divided into multiple mixing cycles and magnetic absorption cycles, and sequentially sorted and completed in the scheduling module. At the same time, the liquid injection can also be performed in sequence in the scheduling module according to the number of liquid injections, realizing the free configuration of process parameters such as reaction steps, liquid addition order, mixing time, magnetic absorption time, etc., and can be compatible with magnetic extraction of solid phase magnetic beads and immunomagnetic beads at the same time, with greater freedom and adaptability, greatly expanding the methodological compatibility.

[0075] Furthermore, the scheduling module includes a liquid injection scheduling module, a mixing scheduling module and a magnetic attraction scheduling module;

[0076] The injection scheduling module is used to add reaction reagents to the reaction cup, and can select the matching number of injection scheduling holes according to the number of injections required for sample pretreatment in the reaction cup. Each time the reaction cup completes an injection and subsequent operations (including mixing, magnetic attraction, and waste liquid extraction), it will be automatically moved forward by one scheduling hole for the next injection until all injections are completed. When the previous reaction cup completes an injection and subsequent operations and is moved forward to the next scheduling hole, the vacant scheduling hole will be filled by the next reaction cup, so the adjacent reaction cups are moved forward in sequence for assembly line operation, and the adjacent reaction cups differ by one injection and subsequent operation process.

[0077] The mixing scheduling module divides the mixing time into multiple reaction cycles for mixing the solution in the reaction cup, and can select the matching number of mixing scheduling holes according to the mixing time required for sample pretreatment in the reaction cup. Each time the reaction cup completes a mixing cycle, it will be automatically moved forward by one scheduling hole to perform the next mixing cycle until all mixing cycles are completed. When the previous reaction cup completes a mixing cycle and is moved forward to the next scheduling hole, the vacant scheduling hole will be filled by the next reaction cup, so the adjacent reaction cups are moved forward in sequence for assembly line operation, and the time difference between adjacent reaction cups is one mixing cycle.

[0078] The magnetic attraction scheduling module divides the magnetic attraction time into multiple reaction cycles, which are used to adsorb the magnetic beads in the reaction cup, and can select the matching number of magnetic attraction scheduling holes according to the magnetic attraction time required for sample pretreatment in the reaction cup. Each time the reaction cup completes a magnetic attraction cycle, it will be automatically moved forward by one scheduling hole to perform the next magnetic attraction cycle until all magnetic attraction cycles are completed. When the previous reaction cup completes a magnetic attraction cycle and is moved forward to the next scheduling hole, the vacant scheduling hole will be filled by the next reaction cup, so the adjacent reaction cups are moved forward in sequence for assembly line operation, and the time difference between adjacent reaction cups is one magnetic attraction cycle.

[0079] The magnetic bead sample pretreatment instrument provided by the present invention sets scheduling modules respectively according to the core parameters of the entire process: the number of reagent injections, mixing time and magnetic absorption time, and sets the three scheduling modules in one area, which are uniformly scheduled in the area by a robotic arm.

[0080] Furthermore, each scheduling hole position of the liquid injection scheduling module can inject reaction reagents into the reaction cup therein; each scheduling hole position of the mixing scheduling module can provide the same-frequency mixing oscillation for the reaction cup therein; and each scheduling hole position of the magnetic attraction scheduling module can put the sample therein into a magnetic attraction state.

[0081] Existing magnetic bead sample pretreatment instruments all require the reaction cup to be grasped by a gripper to a specific position for magnetization and waste liquid absorption. The magnetization module is an integrated structure and cannot be moved to other positions. The magnetization scheduling module provided by the present invention directly sets a magnet at each scheduling hole position, which can enable each sample to directly absorb magnetism for a cycle (such as 1 minute) at each scheduling hole position, and can be moved to the next scheduling hole position in turn to continue magnetization. After the magnetization is completed, the reaction cup and the scheduling hole position move directly to the waste liquid extraction module to absorb the waste liquid. After absorbing the waste liquid, the reaction cup moves to the next process, and the remaining reaction cups and scheduling holes return to the track where the magnetization scheduling module is located to continue the magnetization treatment. In other words, the pretreatment instrument provided by the present invention can combine the two tasks of transportation and magnetization, so that the magnetization time of each sample can be more flexibly controlled, and the magnetization and waste liquid absorption treatment can be completed in a progressive manner one by one in an assembly line.

[0082] Furthermore, the injection scheduling module, mixing scheduling module and magnetic attraction scheduling module are all provided with reaction cup racks, namely the first reaction cup rack, the second reaction cup rack and the third reaction cup rack; the scheduling holes are all located on the reaction cup racks; and each scheduling hole of the third reaction cup rack is provided with a magnet.

[0083] Furthermore, the reaction reagents include one or more of a magnetic bead solution, a washing solution, and an eluting solution; the number of scheduling holes in the reaction cup rack is set according to the longest mixing time, the longest magnetic attraction time, or the maximum number of injections required for sample pretreatment, and each reaction cup represents one injection, or one reaction cycle of mixing, or one reaction cycle of magnetic attraction.

[0084] It is understandable that the number of scheduling holes can be set according to the needs of product pre-processing parameters.

[0085] In some embodiments, the number of scheduling holes on the reaction cup rack is 7, which means that each sample can be injected up to 7 times, mixed up to 7 cycles (such as 1 minute per cycle, the longest mixing time is 7 minutes), and magnetic attraction up to 7 cycles (such as 1 minute per cycle, the longest magnetic attraction time is 7 minutes).

[0086] Furthermore, it also includes a waste liquid extraction module. The reaction cup of the magnetic attraction scheduling module can be transferred as a whole to the waste liquid extraction module, and the waste liquid in the reaction cup under the magnetic attraction state is extracted through the waste liquid needle; the waste liquid extraction module is provided with multiple waste liquid needles and a suction pump, and the suction pump can control the start and stop of each waste liquid needle separately.

[0087] In some embodiments, the number of waste liquid needles of the waste liquid extraction module is consistent with the number of scheduling holes of the third reaction cup rack (magnetic attraction scheduling module).

[0088] In some methods, the number of waste liquid needles in the waste liquid extraction module is 7, and the number of scheduling holes in the third reaction cup rack (magnetic absorption scheduling module) is 7. When the third reaction cup is transferred to the waste liquid extraction module, the 7 waste liquid needles are inserted into each scheduling hole at the same time, but only the waste liquid needle corresponding to the sample that has completed the magnetic absorption time absorbs waste liquid, and the other waste liquid needles are in the pause working stage. This process is completed by the suction pump control. The suction pump can control the 7 waste liquid needles separately to distinguish which waste liquid needle can absorb waste liquid and which waste liquid needle needs to be paused.

[0089] Furthermore, the running track of the magnetic attraction scheduling module is extended to the waste liquid extraction module, so that the reaction cup rack in the magnetic attraction scheduling module can be quickly moved to the waste liquid extraction module.

[0090] The running tracks of the magnetic attraction scheduling module and the waste liquid extraction module are on the same track. The reaction cup holder of the magnetic attraction scheduling module can be moved to the waste liquid extraction module as a whole by simply pushing the reaction cup holder of the magnetic attraction scheduling module, truly combining the two tasks of transportation and magnetic attraction, efficiently completing the magnetic attraction and waste liquid extraction steps, and allowing samples at different magnetic attraction time schedules to complete magnetic attraction and waste liquid extraction in sequence in the assembly line.

[0091] Furthermore, the scheduling module also includes a three-axis robotic arm gripper and an injection robotic arm. The three-axis robotic arm gripper is used to complete the scheduling of the reaction cup between the injection scheduling module, the mixing scheduling module and the magnetic attraction scheduling module; the injection robotic arm is used to complete the injection of the reaction cup in the injection scheduling module.

[0092] The three-axis robotic arm gripper can realize x-, y-, and z-axis movement and can cover the entire scheduling module. It can control the movement of the reaction cup in any scheduling hole position within the scheduling module area. The gripper component can grab a single reaction cup and place it in the appropriate hole position.

[0093] The liquid injection robot arm is used to inject liquid into the reaction cup in the liquid injection scheduling module. The liquid injection robot arm is equipped with a pipette needle, which can transfer the required reagent from the reagent storage bin and inject it into the corresponding reaction cup.

[0094] In some embodiments, the number of the liquid injection robotic arms is two.

[0095] It takes 15 seconds for the injection robot to perform one injection, including the pipette needle sucking up the reagent, injecting into the reaction cup, and cleaning the pipette needle; therefore, when one mixing cycle and magnetic attraction cycle is 1 minute, 4 injections can be completed within the cycle. The scheduling module provided by the present invention is provided with two injection robot arms, so that the injection of 8 reaction cups can be completed together within one cycle. Therefore, as long as the number of scheduling holes in the injection scheduling module does not exceed 8, the two injection robot arms can complete the injection of all the scheduling holes in the injection scheduling module.

[0096] Furthermore, the liquid injection scheduling module, the mixing scheduling module and the magnetic attraction scheduling module are arranged in three rows in parallel.

[0097] When the liquid injection scheduling module, the mixing scheduling module and the magnetic attraction scheduling module are arranged in three parallel rows, the three-axis robotic arm gripper can conveniently determine the specific position of the reaction cup by adjusting the x and y axes, and then move up and down through the z axis to realize the grabbing and selecting the position of the reaction cup in the three scheduling modules, which is more convenient and ensures accuracy.

[0098] Furthermore, it also includes an injection module, which includes a series sample module and an emergency sample module. The series sample module and the emergency sample module differ in one or more of the number of injections, mixing time, and magnetic absorption time required for the pretreatment of samples. The number of the series sample modules or the emergency sample modules is one or more.

[0099] Multiple different series of sample modules can be set in the injection module. The different series of samples refer to samples with different pre-processing process parameters, including one or more of the injection times, mixing time, and magnetic absorption time. By placing them in different series of sample modules, it is easy to distinguish them. For example, when manual placement is required, it can be placed more clearly and intuitively to avoid mistakes.

[0100] In some methods, samples of the same series of sample modules can also be placed according to the position number to distinguish samples of different pre-treatment processes by setting specific position numbers. However, placing samples according to position numbers will increase the probability of error when placing samples manually, compared with placing samples by area. Therefore, it is preferred to place samples in different series of sample modules separately.

[0101] In some methods, solid phase magnetic bead pre-treatment products and immunomagnetic bead pre-treatment products can also be divided into different series of products.

[0102] In some embodiments, the number of the series sample modules or emergency sample modules can be selected according to product needs. For example, if the product to be tested often needs to adjust the pre-treatment method, more series sample modules can be set for differentiation; if there are often different batches of emergency samples that need to be tested, multiple emergency sample modules can be set.

[0103] In some embodiments, the sampling module is provided with a sample pipetting robot, which has the function of transferring samples with high precision as required. The sampling range covers the front fan-shaped area, and the sampling points can be flexibly set, providing conditions for connecting to external production lines.

[0104] Furthermore, the magnetic bead method sample pretreatment instrument also includes a reaction cup and tip head storage scheduling module and a reagent storage compartment module.

[0105] The reaction cup and Tip head storage scheduling module has the functions of temporarily storing Tip heads and reaction cups, loading new Tip heads onto the sample transfer robot of the injection module, loading new reaction cups onto the reagent liquid addition platform, and discarding waste reaction cups.

[0106] The reagent storage compartment module has the functions of reagent storage, reagent information scanning, reagent management and scheduling, magnetic bead solution mixing, and reagent tray constant temperature control.

[0107] Furthermore, it also includes a pipeline docking module, which includes a pipette needle arm, a 96-well plate and / or a single tube, and is used to transfer the pre-treated samples to the single tube or 96-well plate to be loaded and tested, for docking with subsequent high-throughput detection.

[0108] In some methods, the assembly line docking module outputs samples according to the reaction cycle and can be directly adapted to the latest fully automatic single-tube online detection equipment to realize single-tube online enrichment detection; the effective solution in the reaction cup can also be transferred to a 96-well plate through a pipette needle arm, and then offline enrichment detection can be performed, which greatly improves the versatility of the equipment.

[0109] In summary, the present invention synchronously integrates all the elements such as the loading module, reagent storage cabin, liquid injection scheduling module, mixing scheduling module, magnetic absorption scheduling module, waste liquid extraction module, mechanical gripper, and assembly line docking module into the magnetic bead sample pretreatment instrument, and then realizes on-demand caching and intelligent step-by-step progressive scheduling through the intelligent system, thereby achieving the effect of free configuration of process parameters in the whole process; at the same time, each workstation operates in parallel, so that samples with different magnetic bead pretreatment parameters can still be smoothly docked with the assembly line, thereby ensuring the overall high efficiency of the magnetic bead pretreatment.

[0110] By using the magnetic bead sample pretreatment instrument and pretreatment method provided by the present invention, it is possible to adjust the process parameters in a pipeline-type cyclic progressive manner, so that only the cycle time from the beginning to the end of the pretreatment of the first sample is affected, and the subsequent samples still produce one sample per reaction cycle, thereby ensuring the overall high efficiency of the pretreatment equipment.

[0111] The present invention has the following beneficial effects:

[0112] (1) By dividing the mixing time and magnetic attraction time in the sample magnetic bead method pretreatment process into multiple mixing cycles and magnetic attraction cycles, the mixing and magnetic attraction are completed in sequence, and the injection is completed in sequence according to the required number of injections, so that each reaction cup can be cached and intelligently scheduled in the injection scheduling module, mixing scheduling module, and magnetic attraction scheduling module, so that the process parameters such as the number of reagent injections, mixing time, and magnetic attraction time can be freely configured (theoretically supporting the configuration of any process parameters);

[0113] (2) Through the pipeline-style cyclic progressive method, after adjusting the process parameters, only the cycle time from the beginning to the end of the pretreatment of the first sample is affected. The subsequent samples still produce one sample per reaction cycle, ensuring the overall efficiency of the pretreatment equipment;

[0114] (3) A magnetic bead sample pretreatment instrument was designed that can realize the function of scheduling parameters for the pretreatment process. Through the design and arrangement of the structure, the multi-station cache and intelligent scheduling design of the reaction tube, the free configuration of process parameters such as reaction steps, injection sequence, mixing time, and magnetic absorption time was realized, which greatly expanded the methodological compatibility;

[0115] (4) The magnetic bead method sample pretreatment instrument is equipped with a liquid injection scheduling module, a mixing scheduling module, and a magnetic attraction scheduling module, and is arranged in three parallel rows to improve the accuracy and efficiency of scheduling. The three-axis robotic arm can cover the entire scheduling module and can control the movement of the reaction cup in any scheduling hole position within the scheduling module area. An appropriate number of liquid injection needles are set so that the liquid injection process is completed within the mixing cycle or the magnetic attraction cycle without affecting the overall rhythm.

[0116] (5) A magnet is set at each scheduling hole of the magnetic absorption scheduling module, so that each sample can be directly magnetized at each scheduling hole. The running track of the magnetic absorption scheduling module and the waste liquid extraction module are on the same track. After the magnetic absorption is completed, the reaction cup and the scheduling hole are directly moved to the waste liquid extraction module to absorb the waste liquid. The two tasks of transportation and magnetic absorption are combined together, and the magnetic absorption time of each sample can be more flexibly controlled;

[0117] (6) Single-tube reaction cups are used as reaction consumables to achieve immediate processing of samples. A minimum of one sample can be processed on the machine, breaking the disadvantage of the traditional 96-well plate model where the entire plate of samples must be processed together. Single-tube loading and emergency queue jumping are supported, and the process parameters of sample pretreatment can be freely configured to meet the needs of more complex usage scenarios.

[0118] (7) The assembly line operation mode is adopted to realize the parallel operation of each process at different workstations, which greatly increases the throughput of pre-treatment, breaks the traditional pre-treatment series single-line mode, and is more efficient;

[0119] (8) The setting of sampling points is more flexible, which enables the equipment to be used in both independent sampling mode and external assembly line sampling mode, thus expanding the applicable scenarios of pre-processing;

[0120] (9) The magnetic bead sample pretreatment instrument is compatible with magnetic extraction of both solid phase magnetic beads and immunomagnetic beads, and has greater methodological freedom and adaptability;

[0121] (10) A pipetting arm assembly is added to the back end, which can realize single-tube online enrichment detection; the effective solution of the reaction tube can also be transferred to a 96-well plate for offline enrichment detection, so that the device can not only adapt to the latest fully automatic single-tube online detection equipment, but also adapt to conventional 96-well plate detection equipment, greatly improving the versatility of the equipment;

[0122] (11) The whole process is in assembly line operation mode, which only requires manual placement of samples and consumables, and other actions are completed automatically by the equipment; it realizes a fully automated mode of sample input and effective solution output. BRIEF DESCRIPTION OF THE DRAWINGS

[0123] Figure 1 This is a basic reaction flow chart of the magnetic bead method sample pretreatment instrument of Example 2;

[0124] Figure 2 This is a reaction flow chart of a mixing scheduling module of a magnetic bead method sample pretreatment instrument in Example 2;

[0125] Figure 3 This is a reaction flow chart of the magnetic attraction scheduling module of the magnetic bead method sample pretreatment instrument in Example 2;

[0126] Figure 4 The following is a detailed reaction flow chart of the scheduling module of the magnetic bead method sample pretreatment instrument in Example 2;

[0127] Figure 5 It is a top view of the magnetic bead method sample pretreatment instrument of Example 1;

[0128] Figure 6 This is a schematic diagram of the overall structure of the magnetic bead method sample pretreatment instrument of Example 1;

[0129] Figure 7 This is a schematic diagram of the scheduling module structure of the magnetic bead method sample pretreatment instrument in Example 1;

[0130] Figure 8 This is a schematic diagram of the scheduling module structure of the magnetic bead method sample pretreatment instrument in Example 1;

[0131] Fig. 9 This is a schematic diagram of the structure of the magnetic attraction scheduling module of the magnetic bead method sample pretreatment instrument in Example 1;

[0132] Fig.10 This is a schematic diagram of the structure of the three-axis motion mechanism of the magnetic bead method sample pretreatment instrument in Example 1;

[0133] Fig.11 This is a schematic diagram of the structure of a mixing and scheduling module of a magnetic bead method sample pretreatment instrument in Example 1;

[0134] Fig.12 This is a schematic diagram of the longitudinal section structure of the mixing and scheduling module of the magnetic bead method sample pretreatment instrument in Example 1;

[0135] Fig.13 This is a schematic diagram of the structure of the waste liquid extraction module of the magnetic bead method sample pretreatment instrument in Example 1;

[0136] Fig.14 This is a schematic diagram of the structure of the liquid injection mechanical arm of the magnetic bead method sample pretreatment instrument in Example 1;

[0137] Fig.15 This is a schematic diagram of the sample tray structure of the magnetic bead method sample pretreatment instrument in Example 1;

[0138] Fig.16 This is a schematic diagram of the structure of the sample tray module of the magnetic bead method sample pretreatment instrument in Example 1;

[0139] Fig.17 This is a schematic diagram of the structure of the sample pipetting robot of the magnetic bead method sample pretreatment instrument in Example 1;

[0140] Fig.18 This is a schematic diagram of the structure of the reaction cup and tip head storage scheduling module of the magnetic bead method sample pretreatment instrument in Example 1;

[0141] Fig.19 A top view of a reaction cup and a Tip head storage scheduling module of the magnetic bead method sample pretreatment instrument in Example 1;

[0142] Fig. 20 This is a schematic diagram of the structure of the reagent storage compartment module of the magnetic bead method sample pretreatment instrument in Example 1;

[0143] Fig.21 This is a schematic diagram of the structure of the constant temperature refrigeration component of the magnetic bead method sample pretreatment instrument in Example 1;

[0144] Fig. 22 This is a schematic diagram of the structure of the plate transfer and liquid transfer robot of the magnetic bead method sample pretreatment instrument in Example 1. DETAILED DESCRIPTION

[0145] The present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be pointed out that the embodiments described below are intended to facilitate the understanding of the present invention and do not have any limiting effect on the present invention.

[0146] like Figure 5 , 6 As shown, the magnetic bead sample pretreatment instrument 1 provided in this embodiment comprises Figure 5 The dotted frame includes the injection module 2, the reaction cup and tip head storage scheduling module 3, the reagent storage compartment module 4 and the scheduling module 5.

[0147] like Figure 7-8The magnetic bead sample pretreatment instrument 1 provided in this embodiment includes the following three items: the number of times of adding reagents, the mixing time and the magnetic absorption time, according to the core parameters of the entire magnetic bead pretreatment process: Figure 7 , 8 The dotted box marks the liquid injection scheduling module 6, the mixing scheduling module 7 and the magnetic attraction scheduling module 8, and the three scheduling modules are set in one area, and are uniformly scheduled in the area by the three-axis mechanical arm 12. The liquid injection scheduling module 6 is used to add reaction reagents to the reaction cup, and can automatically place the reaction cup in a suitable liquid injection scheduling hole position 9 according to the number of liquid injections required for sample pretreatment in the reaction cup, and each liquid injection scheduling hole position 9 of the liquid injection scheduling module 6 can inject reaction reagents into the reaction cup therein; the mixing scheduling module 7 is used to mix the solution in the reaction cup, and can automatically place the reaction cup in a suitable mixing scheduling hole position 10 according to the mixing time required for sample pretreatment in the reaction cup, and each mixing scheduling hole position 10 of the mixing scheduling module 7 can provide the same frequency mixing oscillation for the reaction cup therein; the magnetic attraction scheduling module 8 is used to adsorb the magnetic beads in the reaction cup, and can automatically place the reaction cup in a suitable magnetic attraction scheduling hole position 11 according to the magnetic attraction time required for sample pretreatment in the reaction cup, and each magnetic attraction scheduling hole position 11 of the magnetic attraction scheduling module 8 can put the sample therein in a magnetic attraction state. The reaction cup is a single-tube test tube. The sample is sucked by the injection module 2 and added to the reaction cup and then enters the scheduling module 5 for processing.

[0148] like Figure 7-8 The liquid injection scheduling module 6, the mixing scheduling module 7 and the magnetic attraction scheduling module 8 are all provided with reaction cup racks, namely the first reaction cup rack 13, the second reaction cup rack 14 and the third reaction cup rack 15; the scheduling holes are all located on the reaction cup racks; and each scheduling hole of the third reaction cup rack 15 is provided with a magnet 16.

[0149] The existing magnetic bead sample pretreatment instruments all require the reaction cup to be grabbed by a gripper to a specific position for magnetization and waste liquid absorption. The magnetization module is an integrated structure and cannot be moved to other positions. The magnetization scheduling module 8 in this embodiment ( Fig. 9), including a third reaction cup rack 15, a driving motor and a track 17, a magnet 16 is arranged on each magnetic absorption scheduling hole position 11, so that each sample can be directly subjected to magnetic absorption for one cycle (such as 1 minute) at each magnetic absorption scheduling hole position 11, and can be moved to the next magnetic absorption scheduling hole position 11 in sequence through the gripper of the three-axis mechanical arm 12 to continue magnetic absorption, after the magnetic absorption is completed, the reaction cup and the third reaction cup rack 15 are directly moved to the waste liquid extraction module 18 to absorb the waste liquid, and the reaction cup after absorbing the waste liquid is moved to the next process through the gripper 19 of the three-axis mechanical arm 12, and the remaining third reaction cup racks 15 and the remaining reaction cups are returned to the position of the magnetic absorption scheduling module 8 along the track 17 to continue the magnetic absorption treatment, that is, the pre-treatment instrument provided by the present invention can combine the two tasks of carrying and magnetic absorption, so that the magnetic absorption time of each sample can be more flexibly controlled, and the magnetic absorption and waste liquid absorption treatment can be completed one by one in a progressive manner of an assembly line.

[0150] like Figure 8 , 10 As shown, the injection scheduling module 6 injects liquid into the reaction cup through the injection mechanical arm 20, including a first reaction cup rack 13, a driving motor and an injection track 21. The first reaction cup rack 13 is provided with a plurality of injection scheduling holes 9. The first reaction cup rack 13 can move on the injection track 21. When the sample needs to be loaded, it moves to the vicinity of the injection module 2. When the liquid needs to be injected, it moves to the vicinity of the reagent storage compartment module 4. After the reaction cup on the injection scheduling hole 9 is injected, it can be moved to the next process (mixing) by the gripper 19 of the three-axis mechanical arm 12.

[0151] like Fig.11 , 12 As shown, the mixing scheduling module 7 is composed of a mixing mounting frame 22 and a mixing component 23; wherein the mixing component 23 mainly includes a mixing motor 24, a mixing cam 25, a second reaction cup rack 14 and a photoelectric sensor 26. The mixing motor 24 is used to drive the mixing component 23 to mix, and the mixing cam 25 is used to improve the mixing effect of the sample in the reaction cup. A plurality of mixing scheduling holes 10 are provided on the second reaction cup rack 14. The photoelectric sensor 26 is used to detect the position of the mixing origin. After the reaction cup on the mixing scheduling hole 10 is mixed, it can be moved to the next process (magnetic attraction) by the gripper 19 of the three-axis robot arm 12.

[0152] The reaction reagents injected by the injection scheduling module 6 include one or more of a magnetic bead solution, a washing solution, and an eluent; the number of scheduling holes in each reaction cup rack is set according to the longest mixing time, the longest magnetic absorption time, or the maximum number of injections required for sample pretreatment, and each reaction cup represents one injection, or a mixing of a reaction cycle, or a magnetic absorption of a reaction cycle. In this embodiment, the number of scheduling holes on each reaction cup rack is 7, which is equivalent to a maximum of 7 injections per sample, a maximum of 7 cycles of mixing (such as 1 minute per cycle, the longest mixing time is 7 minutes), and a maximum of 7 cycles of magnetic absorption (such as 1 minute per cycle, the longest magnetic absorption time is 7 minutes). In general, in the existing magnetic bead method treatment, the mixing and magnetic absorption time do not exceed 7 minutes, and the number of injections will not exceed 7 times. Therefore, such a setting can meet the parameter configuration of any existing magnetic bead method treatment. Of course, the number of scheduling holes can be set according to the needs of the product pretreatment parameters. If it is required to exceed 7 or be less than 7, it can be set accordingly.

[0153] The scheduling module 5 uses the gripper 19 of the three-axis robot arm 12 to grab the reaction cup and schedule it in the area. Fig.10 The three-axis robot arm 12 includes a horizontal X-axis driving component 27, a Y-axis driving component 28, and a Z-axis driving component 29, so as to realize movement in the X, Y, and Z axes, and can cover the entire scheduling module. The gripper 19 is a rotating G-axis reaction cup scheduling gripper. The gripper 19 is used to flexibly grasp the reaction cup and place it in a suitable hole position, and the gripper 19 is scheduled to operate at any point within the area.

[0154] The running track 17 of the magnetic attraction scheduling module 8 is extended to the waste liquid extraction module 18, that is to say, the running tracks 17 of the magnetic attraction scheduling module 8 and the waste liquid extraction module 18 are on the same track. It only needs to drive the third reaction cup rack 15 of the magnetic attraction scheduling module 8 to quickly move the third reaction cup rack 15 as a whole to the waste liquid extraction module 18, thereby efficiently completing the magnetic attraction and waste liquid extraction steps, truly combining the two tasks of transportation and magnetic attraction, so that samples at different magnetic attraction time progress can complete magnetic attraction and waste liquid extraction in sequence in the assembly line.

[0155] In the scheduling module 5, the injection scheduling module 6, the mixing scheduling module 7 and the magnetic attraction scheduling module 8 are arranged in three parallel rows. The three parallel rows can facilitate the gripper 19 to determine the specific position of the reaction cup by adjusting the X and Y axes, and then move up and down through the Z axis to realize the grabbing and selecting the position of the reaction cup in the three scheduling modules, which is more convenient and ensures accuracy.

[0156] like Figure 8The waste liquid extraction module 18 marked by the dotted line frame is equipped with a suction pump and multiple waste liquid needles to extract the waste liquid in the reaction cup under the magnetic state; the waste liquid extraction module is equipped with multiple waste liquid needles and a suction pump, and the suction pump can control the start and stop of each waste liquid needle separately. Fig.13 , the waste liquid extraction module 18 mainly includes a drive motor and a transmission component 30, a cam lifting slider 31, a waste liquid needle mounting seat 32, a waste liquid needle 33, a waste liquid needle mounting seat 34, and a waste liquid needle anti-collision spring 35. The number of waste liquid needles 33 of the waste liquid extraction module 18 is consistent with the number of scheduling holes of the third reaction cup rack 15. The number of waste liquid needles 33 of the waste liquid extraction module 18 is 7, and the number of scheduling holes of the third reaction cup rack 15 is 7. When the third reaction cup rack 15 is transferred to the waste liquid extraction module 18, the 7 waste liquid needles 33 are inserted into each scheduling hole at the same time, but only the waste liquid needle corresponding to the sample that has completed the magnetic absorption time absorbs waste liquid, and the other waste liquid needles are in the pause working stage. This process is completed by the control of the suction pump. The suction pump can control the 7 waste liquid needles separately to distinguish which waste liquid needle can absorb waste liquid and which waste liquid needle needs to be paused.

[0157] The injection scheduling module 6 in the scheduling module 5 requires the injection robot 20 to complete the injection of the injection scheduling module 6. Figure 8 , 14 As shown, the liquid injection robot 20 is mainly composed of a needle arm mounting support 36, a vertical drive motor and transmission component 37, a rotary drive motor and transmission component 38, a needle arm 39, a pipette needle 40 and other components. There are two liquid injection robot arms 20, which can remove the required reagents from the reagent storage bin and inject them into the corresponding reaction cup. It takes a total of 15 seconds for the liquid injection robot arm 20 to perform one injection, including the pipette needle absorbing the reagent, injecting the reaction cup, and cleaning the pipette needle 40; therefore, when a mixing cycle and a magnetic attraction cycle are 1min, 4 injections can be completed within the cycle. The scheduling module 5 provided by the present invention is provided with two liquid injection robot arms, so the injection of 8 reaction cups can be completed together within one cycle. Therefore, as long as the number of scheduling holes in the liquid injection scheduling module 6 does not exceed 8, the two liquid injection robot arms can complete the injection of all the scheduling holes in the liquid injection scheduling module 6.

[0158] like Figure 6 The sample injection module 2 shown in FIG. 4 includes a sample tray module 41 and a sample transfer robot 42. Fig.15 , 16As shown, the sample tray module 41 includes a sample tray bottom bracket 43, a sample tray outer tray 44, a barcode scanner component 45, a drive motor and transmission component 46, and a sample tube basket component 47. The sample tray bottom bracket 43 is used to support and install the entire sample tray assembly, and the sample tray outer tray 44 also serves as an overall frame to provide an installation basis for other structures. The barcode scanner component 45 is mainly used for inputting and identifying the barcode information of the sample tube, etc. The drive motor and transmission component 46 are mainly used to drive the rotating shaft and drive the sample basket component 47 to achieve the scheduling and management of sample sampling at different positions. The sample basket component 47 is mainly used to load the sample tubes to be tested. The entire sample basket component 47 has 3 layers in total. The outer two layers are series sample modules 48 with a total of 40 spaces for placing conventional samples to be tested, and the innermost layer is an emergency sample module 49 with a total of 10 spaces for placing urgent samples. When the entire equipment is connected to an external assembly line, the emergency sample module 49 can be used to support the emergency sample queue-jumping function. Fig.17 As shown, the sample pipetting robot 42 includes a three-axis mechanical arm 50 and a single-pass pipette 51. The three-axis mechanical arm 50 can realize effective operation within a large range of fan-shaped areas, realize flexible setting of sample sampling points, and support the connection of external production lines; the single-pass pipette 51 can realize high-precision sample pipetting and has comprehensive use functions such as Tip head detection, liquid level detection, and anti-blocking detection.

[0159] The reaction cup and tip head storage scheduling module 3 has the functions of temporarily storing the tip head and reaction cup, loading the new tip head onto the sample transfer robot of the injection module, loading the new reaction cup onto the reagent liquid loading platform, and discarding the waste reaction cup. Fig.18 , 19 As shown, the reaction cup and tip head storage scheduling module 3 includes a frame support limiting mechanism 52, a vertical Y-axis drive mechanism 53, a horizontal X-axis drive mechanism 54, a vertical Z-axis drive mechanism 55, a rotating axis gripper mechanism 56, and a tip head and reaction cup placement carrier 57. The frame support limiting mechanism 52 is mainly used for structural installation, carrier placement and limiting functions; the vertical Y-axis drive mechanism 53, the horizontal X-axis drive mechanism 54, and the vertical Z-axis drive mechanism 55 are three-axis motion mechanisms, so that the gripper can reach any point in the area to operate; the rotating axis gripper mechanism 56 is mainly used for grabbing the tip head and reaction cup; the tip head and reaction cup placement carrier 57 is mainly used for loading and placing the tip head and reaction cup. The sample pipetting robot 42 injects the sample in the sample tray module 41 into the reaction cup placed on the tip head and reaction cup placement carrier 57, and the rotating axis gripper mechanism 56 grabs the reaction cup and moves it through the X, Y, and Z three-axis motion mechanisms to send it to the first reaction cup rack 13 for the next step (liquid injection).

[0160] The reagent storage module 4 has the functions of reagent storage, reagent information scanning, reagent management and scheduling, magnetic bead solution mixing, reagent tray constant temperature control, etc. Figure 6 , 20 As shown in Figure 21, the reagent storage chamber module 4 includes a reagent chamber body 58 and a constant temperature refrigeration assembly 59. The reagent chamber body 58 includes a reagent tray bracket 60, a drive motor and a transmission assembly 61, a reagent tray body 62, and a barcode scanner component 63; the constant temperature refrigeration assembly 59 includes a refrigeration bracket 64, a ceramic refrigeration plate 65, a cold end heat sink 66, a hot end heat sink 67, a refrigeration end fan 68, a heat dissipation end fan 69, a temperature sensor 70, and a thermal insulation layer 71. Among them, the reagent tray bracket 60 is used for supporting and installing the reagent tray; the drive motor and transmission assembly 61 are used for rotating the inner tray of the reagent, realizing the on-demand scheduling of the reagent kit and the mixing of the magnetic bead solution; the reagent tray body 62 is used for mounting structure and constant temperature insulation; the barcode scanner component 63 is used for loading and unloading the reagent kit, reading and entering the reagent information; the refrigeration bracket 64 is used for installing the refrigeration module; the ceramic refrigeration plate 65 is used for refrigeration; the cold end heat sink 66 is used for low temperature conduction; the hot end heat sink 67 is used for heat extraction; the refrigeration end fan 68 is used to introduce the low temperature of the cold end heat sink into the reagent tray; the heat dissipation end fan 69 is used to discharge heat into the air; the temperature sensor 70 is used to detect the real-time temperature in the tray; the insulation layer 71 is used for insulation.

[0161] like Figure 6 As shown, the pipeline docking module 81 includes a pipette needle arm 72 and a 96-well plate 82 (or a single tube). The sample that has completed the pre-treatment is sampled out according to the reaction cycle through the pipeline docking module 81. The effective solution in the reaction cup can be transferred to the single tube through the pipette needle arm 72, which is directly adapted to the latest fully automatic single-tube online detection equipment to achieve single-tube online enrichment detection; the effective solution in the reaction cup can also be transferred to the 96-well plate through the pipette needle arm 72, and then offline enrichment detection is performed, which greatly improves the versatility of the equipment. Fig.18 As shown, the pipetting needle arm 72 includes a double-axis needle arm mounting seat 73, a vertical drive motor and a transmission component 74, a rotation drive motor and a transmission component 75, a needle arm drive motor and a transmission component 76, a buffer anti-collision spring 77, a first needle arm 78, a second needle arm 79, and a rotating plate needle 80. The double-arm structure design of the first needle arm 78 and the second needle arm 79 can greatly increase the area covered by the rotating plate needle 80 for pipetting.

[0162] 1. Pipeline Scheduling Method

[0163] In order to realize the free configuration of process parameters such as the number of injection reagent types, mixing time, magnetic attraction time, etc., in addition to the above-mentioned structural design and implementation, it is also necessary to reasonably design and optimize the scheduling of reaction cups at each station.

[0164] like Figure 1, which represents the most basic processing reaction process, namely reagent injection - mixing - magnetic attraction - waste liquid extraction - elution; following this process can achieve the simplest pre-treatment process, but one must wait for a sample to complete the entire process of reagent injection - mixing - magnetic attraction - waste liquid extraction - elution before the next sample can be processed. The methodology is extremely fixed and rigid, and lacks adjustability and adaptability.

[0165] In actual use, different projects may have different requirements for mixing time. For example, project A requires 1 mixing cycle, project B requires 2 mixing cycles, and project N requires n mixing cycles. To adapt to this situation, this example 2 uses the cache design of the magnetic carrier and the step-by-step scheduling of the reaction cup to meet the requirements of different projects for different mixing cycles. Figure 2 , the specific steps include:

[0166] (1) Move the injection scheduling module to the sample loading area and load the reaction cup into the A1 injection scheduling hole of the injection scheduling module;

[0167] (2) The injection robot transfers the solution into the reaction cup at the injection scheduling hole A1;

[0168] (3) The gripper grabs the reaction cup that has completed the injection at the A1 injection scheduling hole and moves it to the B1 mixing scheduling hole for mixing;

[0169] (4) The gripper grabs the reaction cup that has completed the mixing cycle at the B1 mixing scheduling hole and moves it to the B2 mixing scheduling hole for mixing;

[0170] ………

[0171] (5) The gripper grabs the reaction cup that has completed the mixing cycle at the B(n-1) mixing scheduling hole and moves it to the Bn mixing scheduling hole for mixing;

[0172] (6) The gripper grabs the Bn mixing scheduling hole to complete the mixing cycle of the reaction cup and moves it to the C1 magnetic attraction scheduling hole for static magnetic attraction;

[0173] (7) The magnetic suction scheduling hole is moved to the waste liquid extraction area to complete the waste liquid extraction;

[0174] (8) Elution injection.

[0175] Similarly, in actual use, different projects have different requirements for magnetization time. For example, project A requires 1 magnetization cycle, project B requires 2 magnetization cycles, and project N requires n magnetization cycles. In order to adapt to this situation, this example 2 also performs a cache design on the magnetization stage and a step-by-step scheduling of the reaction cups, thus achieving the requirements of different projects for different magnetization cycles. Figure 3 , the specific steps include:

[0176] (1) Move the injection scheduling module to the sample loading area and load the reaction cup into the A1 injection scheduling hole of the injection scheduling module;

[0177] (2) The injection robot transfers the solution into the reaction cup at the injection scheduling hole A1;

[0178] (3) The gripper grabs the reaction cup that has completed the injection at the A1 injection scheduling hole and moves it to the B1 mixing scheduling hole for mixing;

[0179] (4) The gripper grabs the reaction cup that has completed the mixing at the B1 mixing and scheduling hole and moves it to the C1 magnetic attraction scheduling hole for static magnetic attraction;

[0180] (5) The gripper grabs the reaction cup that has completed the magnetic attraction at the C1 magnetic attraction scheduling hole and moves it to the C2 magnetic attraction scheduling hole for static magnetic attraction;

[0181] ………

[0182] (6) The gripper grabs the reaction cup that has completed the magnetic attraction cycle at the C(n-1) magnetic attraction scheduling hole and moves it to the Cn magnetic attraction scheduling hole for magnetic attraction;

[0183] (7) The magnetic suction scheduling hole is moved to the waste liquid extraction area to complete the waste liquid extraction;

[0184] (8) Elution injection.

[0185] The cache design of the mixing station and the magnetic attraction station and the progressive scheduling of the reaction cups can effectively solve the problem of free setting of process parameters such as mixing time and magnetic attraction time for different projects. However, in the actual use of pretreatment, there is another very important parameter that needs to be freely configured in terms of method, that is, the number of reagent types that need to be added. For example, project A only needs to add 1 reagent, and after completing the mixing-magnetic attraction-waste liquid extraction process, elution and injection can be carried out; project B needs to add 2 reagents, and after completing the mixing-magnetic attraction-waste liquid extraction process in turn, elution and injection can be carried out; and project N needs to add n reagents, and after completing the mixing-magnetic attraction-waste liquid extraction process in turn, elution and injection can be carried out. In order to realize the free configuration of the number of reagent types, this example 2 carries out a cache design for the liquid injection carrier and a progressive scheduling design for the reaction cups. Figure 4 As shown, the specific steps can be simplified as follows:

[0186] (1) Move the injection scheduling module to the sample loading area and load the reaction cup into the A1 injection scheduling hole of the injection scheduling module;

[0187] (2) The injection robot transfers the solution into the reaction cup at the injection scheduling hole A1;

[0188] (3) The gripper grabs the reaction cup that has completed the injection at the A1 injection scheduling hole and moves it to the B1 mixing scheduling hole for mixing; and moves from B1 to Bx in sequence to complete the mixing of each mixing cycle; when a reaction cup is moved out of the A1, B1~Bx positions, the next reaction cup is immediately added until the mixing of reaction cup an is completed;

[0189] (4) The gripper grabs the reaction cup and moves it from the Bx position to the C1 position of the magnetic attraction scheduling module to perform the first magnetic attraction cycle; and moves from C1 to Cy in sequence to complete the magnetic attraction of each magnetic attraction cycle; when a reaction cup is moved out of the A1, B1~Bx, C1~Cy positions, the next reaction cup is immediately added until the magnetic attraction of reaction cup an is completed;

[0190] (5) After completing the magnetic attraction and the waste liquid, the reaction cup is moved from the Cy position to the A2 position of the injection scheduling module to complete the second injection, and then the mixing and magnetic attraction are completed in accordance with steps (3) and (4) until n injections and the corresponding mixing and magnetic attraction are completed.

[0191] 2. Comparison with commercially available magnetic bead sample pretreatment instruments

[0192] The magnetic bead sample pretreatment instruments available on the market are usually simple serial single-line modes, that is, each step is completed in sequence by different series of samples, and each reaction process cannot occur at the same time. For example, series a samples need to be injected once, mixed for 1 minute, and magnetically attracted for 3 minutes, and series b samples need to be injected once, mixed for 3 minutes, and magnetically attracted for 2 minutes. In this case, you need to set the parameters of series a samples first, complete the pretreatment process of all series a samples, and then set the parameters of series b samples and load the pretreatment process of series b samples. If a series of samples needs to be injected 2 times or more, the mixing-magnetic attraction-waste liquid extraction process needs to be repeated after each injection until the last injection and subsequent operations are completed before the sample is eluted. In the magnetic bead method sample pretreatment instrument of the present invention, the reaction processes of injection, mixing, and magnetic absorption can occur at the same time, and each process module can be synchronized, and the pipeline can be advanced forward in sequence. The parameters can be freely configured. After the parameters are set, each pretreatment process is highly automated, and the pretreatment processes of different series of samples are automatically connected without interruption, which does not affect the overall rhythm and efficiency, thereby ensuring the high efficiency of the pretreatment equipment and breaking the traditional pretreatment series single-line mode. For example, the pretreatment steps and time of the a series of samples are the same, that is, the number of injections and the mixing time and magnetic absorption time after each injection are the same, so only the first sample is sampled from the first injection to the end of the pretreatment, and the sampling frequency of subsequent samples is one sample per reaction cycle. The commercially available magnetic bead method sample pretreatment instrument needs to complete the pretreatment process of all a series of samples, and the samples are sampled at the same time. When switching from a series of samples to b series of samples, the magnetic bead method sample preprocessing instrument of the present invention only has the first b series sample because it needs to increase the injection time once, and adds a round of injection, mixing, magnetic absorption, and waste liquid absorption process, resulting in a delay in the sample output time. Starting from the second b series sample, the original assembly line automation preprocessing rhythm can still be maintained, and one sample is output per reaction cycle. The commercially available magnetic bead method sample preprocessing instrument obtains the b series samples only after completing the preprocessing process of all a series samples, and then completing the preprocessing process of all b series samples, before the b series samples can be sampled, and it cannot be sampled at a certain frequency.

[0193] In summary, the assembly line scheduling method for magnetic bead sample pretreatment provided by the present invention integrates elements such as the injection module, reaction cup and Tip head storage scheduling module, reagent storage compartment module and scheduling module into the pretreatment equipment, and then realizes on-demand caching and intelligent step-by-step progressive scheduling through an intelligent system, thereby achieving the effect of free configuration of process parameters in the entire process; at the same time, each workstation operates in parallel, so that except for affecting the cycle time from the start to the end of pretreatment of the first sample, the subsequent samples still have a frequency of one sample per reaction cycle, thereby ensuring the overall efficiency of the pretreatment equipment.

[0194] Although the present invention is disclosed as above, the present invention is not limited thereto. Any person skilled in the art can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the scope defined by the claims.

Claims

1. A pipeline scheduling method for magnetic bead sample pretreatment, characterized in that: The sample pretreatment includes mixing and magnetic attraction; the mixing is divided into a plurality of mixing cycles according to the mixing time and is completed in sequence, and the mixing cycle is the minimum unit time that can be measured when the sample is mixed; the magnetic attraction is divided into a plurality of magnetic attraction cycles according to the magnetic attraction time and is completed in sequence, and the magnetic attraction cycle is the minimum unit time that can be measured when the sample is magnetically attracted; the sample pretreatment also includes liquid injection; the liquid injection is completed in sequence according to the required number of liquid injections; the sample pretreatment is completed by a magnetic bead method sample pretreatment instrument, and a scheduling module is provided on the magnetic bead method sample pretreatment instrument, and the scheduling module includes a liquid injection scheduling module, a mixing scheduling module and a magnetic attraction scheduling module; the sample pretreatment is completed by a magnetic bead method sample pretreatment instrument, and a scheduling module is provided on the magnetic bead method sample pretreatment instrument, and the scheduling module includes a liquid injection scheduling module, a mixing scheduling module and a magnetic attraction scheduling module; the following steps are included: (1) Set the mixing cycle and magnetic absorption cycle of the sample; (2) According to the mixing time and magnetic absorption time required for the sample pre-treatment by magnetic beads, determine the number of mixing cycles and magnetic absorption cycles required for mixing and magnetic absorption respectively; (3) According to the number of injections, mixing cycles, and magnetic absorption cycles required for the sample to be pre-treated by the magnetic bead method, the number of scheduling holes required for the sample in the injection scheduling module, mixing scheduling module, and magnetic absorption scheduling module should be determined; (4) sequentially completing the processing of the sample at the scheduling well positions within the number of scheduling well positions; The injection scheduling module is used to add reaction reagents to the reaction cup, and can select a matching number of injection scheduling hole positions according to the number of injections required for sample pretreatment in the reaction cup. After each injection and subsequent operations are completed, the reaction cup will be automatically moved forward by one scheduling hole position for the next injection until all injection times are completed; when the current reaction cup is moved forward to the next scheduling hole position after completing one injection and subsequent operations, the vacant scheduling hole position is filled by the next reaction cup; the subsequent operations include mixing, magnetic absorption, and waste liquid extraction.

2. The method according to claim 1, characterized in that The steps of sample pretreatment include: liquid injection-mixing-magnetic attraction-waste liquid attraction; the number of liquid injections includes one or more times, and each time the liquid is injected, it is necessary to mix-magnetic attraction-waste liquid attraction; the sample is placed in a reaction cup; the reaction cup can be moved between different scheduling holes by a mechanical gripper until the pretreatment of the sample is completed.

3. The method according to claim 1, characterized in that One scheduling hole position of the injection scheduling module described in step (3) corresponds to one injection into the reaction cup therein; one scheduling hole position of the mixing scheduling module corresponds to one reaction cup therein completing mixing for one reaction cycle time; one scheduling hole position of the magnetic attraction scheduling module corresponds to one reaction cup therein completing magnetic attraction for one reaction cycle time; after each reaction cup is processed at the scheduling hole position, it is removed for the next step of processing.

4. The method according to claim 1, characterized in that The injection in step (3) is completed by an injection robot, and the time required to complete one injection does not exceed the time of the mixing cycle or the magnetic attraction cycle.

5. The method according to claim 1, characterized in that The samples include a series of samples, and the a series of samples include samples a1, a2~an. The number of injections, mixing time and magnetic absorption time required for the pre-treatment reactions of the samples a1 and a2~an are consistent; the scheduling hole positions of the injection scheduling module include A1, A2~An, the scheduling hole positions of the mixing scheduling module include B1, B2~Bx, and the scheduling hole positions of the magnetic absorption scheduling module include C1, C2~Cy, and n, x, and y are integers greater than or equal to 2, n=the number of injections required for the sample; x=the number of mixing reaction cycles required for the sample; y=the number of magnetic absorption reaction cycles required for the sample; The method comprises the following steps: Ⅰ. Load sample a1 into reaction cup a1 and place it at position A1 of the injection scheduling module to complete the injection; II. Move the reaction cup a1 from A1 to B1 of the mixing scheduling module to perform the mixing of the first mixing cycle; and move it from B1 to Bx in sequence to complete the mixing of each mixing cycle; when a reaction cup is moved out of the A1, B1~Bx positions, immediately add the next reaction cup until the mixing of the reaction cup an is completed; III. Move the reaction cup a1 from the Bx position to the C1 position of the magnetic attraction scheduling module to carry out the first magnetic attraction cycle; and move from C1 to Cy in sequence to complete the magnetic attraction of each magnetic attraction cycle; when a reaction cup is moved out of the A1, B1~Bx, C1~Cy positions, immediately add the next reaction cup until the magnetic attraction of the reaction cup an is completed; V. After completing the magnetic attraction, the reaction cup moves from the Cy position to the A2 position of the injection scheduling module to complete the second injection, and then completes the mixing and magnetic attraction in accordance with steps II and III, until n injections and corresponding mixing and magnetic attraction are completed.

6. The method according to claim 1, characterized in that When the samples also include emergency treatment series samples and / or other series samples, the emergency treatment series samples can be inserted into the a series samples for emergency loading at any time. During the pretreatment process of the emergency treatment series samples and other series samples, they can have one or more of different injection times, different mixing times, or different magnetic absorption times compared with the a series samples.

7. A method for achieving sequential pipeline processing of samples with different pre-treatment parameters and sequential sample output in each reaction cycle, characterized in that: The method according to any one of claims 1 to 6 is used for processing, wherein the reaction period is the minimum unit time that can be measured when the sample is mixed and / or magnetically attracted.

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