A kit instant preparation module and a method for instant preparation thereof

Abstract

The application discloses a kit instant preparation module and a kit instant preparation method thereof, relates to the technical field of automatic processing, and classifies reagents required by each step of an extraction system adaptively, adds a large volume of the reagents by using a pumping and adding unit, and adds the reagents by using a pipetting assembly, so that the configuration of the entire extraction reaction kit is completed. In the configuration process, the kit instant preparation module has a smaller risk of blockage, the prepared extraction reaction kit has more uniform and consistent performance, and is more suitable for the scene requirement of accurate diagnosis of various diseases.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of automation processing, in particular to a real-time chemical kit preparation module and a real-time chemical preparation method thereof. BACKGROUND

[0002] In the modern medical system, accurate and early detection is an important prerequisite for treatment and recovery. As an ex vivo detection scheme, in vitro diagnostic technology is a basic means of accurate and early detection. Molecular diagnosis in in vitro diagnosis is a diagnostic technology that takes nucleic acid fragments as the analysis object. Nucleic acid fragments are the genetic basis of various structural proteins, enzymes, antigens and antibodies, and immunologically active molecules related to diseases. In other words, molecular diagnosis diagnoses from the root of the disease or defect. Therefore, molecular diagnosis is widely used in modern medical systems, disease prevention and control systems, and customs inspection and quarantine systems. Thus, various types of biological samples (sample types include human or animal tissue fluid, amniotic fluid, blood, etc., biological tissue section separation fluid, plant root stem leaf section separation fluid, animal epidermis separation fluid, etc., and respiratory tract intestinal tract collection swab preservation fluid, etc.), once extracted from the patient's body, must be processed in a suitable processing scheme. The processing scheme usually involves using molecular diagnostic techniques to quantitatively or qualitatively analyze target nucleic acid fragments. The preparation of molecular diagnostic samples is a time-consuming and labor-intensive step that does not require highly specialized skills. Subsequent amplification and quantitative and qualitative analysis steps are usually performed by personnel who have been specially trained and can use professional equipment.

[0003] Currently, many devices separate nucleic acid extraction and amplification devices in different areas and are completed by different operators. Although this can use a large amount of manpower to make up for the processing speed of a large sample amount, it has a high risk of operator contamination, and the detection process can introduce subjective factors to cause detection errors, so it is necessary to develop an automatic real-time configuration extraction reaction kit suitable for the nucleic acid extraction process, and to develop an automatic sample processing system, which only needs to add samples, reagents, consumables, and quality control products in batches, and the automatic sample processing system can automatically complete the extraction and amplification detection process, which can well solve the problem of excessive human involvement. Currently, some leading companies such as Roche, Abbott and Beckman have developed integrated high-flow diagnostic systems. These systems basically design a scheme for real-time configuration of extraction and amplification reaction systems using commercial reagents, rather than pre-packaged reagent kits. In order to ensure that the reagents in the extraction reaction kit can be accurately quantified and configured, the scheme authorized by US patent US8951805B2 arranges the liquid reagent containers to be filled in parallel, and can use sensors to monitor the liquid level in the filling container in real time. When it is detected that the liquid volume in the filling container is less than the preset value, a pump can be started to pump the corresponding liquid to be filled in another container into the filling container, which can realize rapid liquid filling to make the entire filling operation more continuous and efficient. The scheme authorized by US patent US8715574B2 focuses on protecting the filling control method of the entire system. Roche has authorized a scheme in China that can automatically fill reagents, in which the liquid reagent to be filled is stored in a large-volume barrel form and can be loaded in batches. A plurality of corresponding vibrating string sensors are configured on the barrel reagent storage site to accurately sense the weight change of the barrel reagent, thereby achieving the goal of accurately controlling the liquid volume of the filled liquid. The reagent storage barrel is improved and designed to better meet the on-site reagent filling requirements. US patent application US20210199682A1 uses a similar weighing method to fill reagents, but the system detection scenario limits the application of the system to white blood cell concentration detection. In order to ensure the uniformity of the filled reagents, some manufacturers disclose a scheme in which a stirring unit is configured in the reagent barrel to make the reagent to be filled more uniform through intermittent or continuous stirring action.

[0004] However, the disclosed scheme is not the best solution for the configuration of the nucleic acid extraction reaction kit, which includes a large difference in the amount of different types of reagents required for different extraction steps, such as the amount of eluent and washing solution is much less than that of the lysis solution, the use of the pumping filling scheme has difficulty in precise control of the amount, the pumping equipment also needs to be more precise, the magnetic bead solution usually contains an outer coating, is more suitable for storage in a concentrated state, and its mixed solution is a solid-liquid two-phase state, the cost of precise pumping filling is higher, and the frequent start-stop of pumping filling can easily cause system blockage, so a low-cost and high-precision instant preparation module suitable for extraction reaction kit and an instant preparation method of the kit are urgently needed to be developed. SUMMARY

[0005] The purpose of the present application is to provide a kit instant preparation module and its instant preparation method, which classifies the reagents required for each step of the extraction system adaptively, uses a pumping filling unit to fill a large volume of part of the reagents, and uses a pipetting assembly to transfer and fill part of the reagents, to complete the configuration of the entire extraction reaction kit. The kit instant preparation module has a lower risk of blockage during configuration, and the prepared extraction reaction kit has more uniform performance and is more suitable for the scene requirements of accurate diagnosis of various diseases.

[0006] The technical scheme adopted by the present application is as follows:

[0007] A kit instant preparation module, comprising a pumping filling assembly and a pipetting assembly, the pumping filling assembly can pump and fill a predetermined volume of to-be-filled reagent into a specified well, the pipetting assembly can transfer a predetermined volume of pipetting reagent into a specified well, and the well to be received by the pipetting reagent contains to-be-filled reagent with a volume greater than the pipetting reagent.

[0008] Further, the pumping filling assembly comprises a plurality of filling units, and the plurality of filling units are in fluid communication with corresponding to-be-filled reagent barrels through corresponding filling pumps.

[0009] Further, it also includes a reagent preparation area, which contains a proteinase K carrying area, an internal reference carrying area and an activation solution preparation area. The pipetting assembly can move between the proteinase K carrying area, the internal reference carrying area and the activation solution preparation area, and transfer proteinase K and internal reference to the activation solution preparation area in a set volume ratio. The pipetting assembly can repeatedly perform liquid suction and liquid discharge operations in the activation solution preparation area to obtain the activation solution by stirring and mixing.

[0010] Further, it also includes a magnetic bead mixed solution oscillation assembly, which carries a magnetic bead mixed solution reagent bottle. The magnetic bead mixed solution oscillation assembly can oscillate and shake the magnetic bead mixed solution reagent bottle to generate a vortex effect to mix the magnetic beads in the magnetic bead mixed solution reagent bottle.

[0011] Further, the pumping and filling assembly is selectively fluidly connected to a cleaning liquid tank or a reagent tank to be filled through a liquid path assembly; further comprising a cleaning tank, which is fluidly connected to the cleaning liquid tank through a cleaning pump; the pumping and filling assembly is movable to the cleaning tank for outer wall cleaning, and is connectable to the cleaning liquid tank for inner wall cleaning.

[0012] Further, the pumping and filling assembly is arranged on a fixed cross beam and is driven to slide along the length direction of the fixed cross beam by a driving motor of the fixed cross beam, and an extraction and preparation position and a cleaning tank are arranged in parallel in the lateral direction below the fixed cross beam, the pumping and filling assembly is driven to be aligned with the extraction and preparation position to complete the filling operation, or is driven to be aligned with the cleaning tank to complete the cleaning operation; the extraction and preparation position is equipped with an extraction and reaction reagent box, a plurality of hole positions are arranged in the extraction and reaction reagent box in the lateral and longitudinal directions, and the extraction and preparation position is driven to move in the longitudinal direction, so that a specified hole position of the extraction and reaction reagent box in the longitudinal direction is aligned with the filling unit of the pumping and filling assembly.

[0013] Further, further comprising a first three-way valve, a second three-way valve, a cleaning two-way valve and a cleaning three-way valve; three branches of the first three-way valve are respectively connected to the reagent tank to be filled, the cleaning liquid tank and the second three-way valve, other two branches of the second three-way valve are respectively connected to the filling pump and the pumping and filling assembly; two branches of the cleaning two-way valve are respectively connected to the cleaning tank and the cleaning three-way valve, and other two branches of the cleaning three-way valve are respectively connected to the cleaning pump and the cleaning liquid tank.

[0014] Further, comprising a reagent tank bearing part, the reagent tank bearing part comprises a plurality of reagent tank receiving positions, each reagent tank receiving position is provided with a corresponding gravity sensing unit, the reagent tank receiving position can bear the reagent tank to be filled, a bottle cap unit capable of lifting motion is arranged above the corresponding reagent tank to be filled, the bottle cap unit can cover the barrel opening of the reagent tank to be filled, a reagent needle capable of connecting the inside of the reagent tank to be filled and the liquid path of the pumping and filling assembly is arranged on the bottle cap unit, and a bottle cap buffer is arranged at the position corresponding to the barrel opening of the reagent tank to be filled, which can buffer and support the descending bottle cap unit.

[0015] A kit instant preparation method, comprising the following steps:

[0016] Pumping step: starting the pumping and filling assembly, pumping a preset volume of reagent to be filled into the specified hole position of the kit;

[0017] Pipetting step: starting the pipetting assembly, transferring a preset volume of pipetting reagent into the specified hole position containing the reagent to be filled, and the preset volume of pipetting reagent is less than the preset volume of the reagent to be filled.

[0018] Further, in the pumping step, the magnetic bead diluent filling part in the pumping filling assembly is started, a second volume of magnetic bead diluent is pumped and filled into the designated hole position in the kit, the lysis solution filling part in the pumping filling assembly is started, and a fourth volume of lysis solution is pumped and filled into the designated hole position in the kit; in the pipetting step, the magnetic bead mixing liquid oscillation assembly is started, and the magnetic bead mixing reagent bottle is shaken to make the magnetic bead mixing liquid in the magnetic bead mixing reagent bottle produce a vortex effect to mix the magnetic beads; the pipetting assembly is started, and the first volume of magnetic bead mixing liquid is transferred from the mixed magnetic bead mixing reagent bottle to the hole position containing the second volume of magnetic bead diluent, and the third volume of activation liquid containing protease K and internal reference is transferred to the hole position containing the fourth volume of lysis solution; wherein the second volume is not less than three times the first volume, and the fourth volume is not less than three times the third volume.

[0019] In summary, due to the adoption of the above technical solutions, the beneficial effects of the present application are:

[0020] 1. The present application configures two types of reagent transfer assemblies, i.e., the pumping filling assembly and the pipetting assembly, to at least partially separate the reagents required for different steps, so that the operator does not need to prepare them in advance, but can directly load different types of reagent containers to different storage positions, and the instant preparation module itself can realize reagent filling of different volumes and different precisions through the pumping filling assembly and the pipetting assembly, which requires less human intervention and does not need special reagent configuration high-precision conveying equipment, and the pumping system also does not need to be cleaned too frequently.

[0021] 2. The pumping filling assembly of the present application can first fill a preset volume of each type of reagent into at least part of the hole positions of the kit, and then use a pipette to transfer another type of reagent into at least part of the hole positions of the kit, so that small-volume reagents can be more uniformly mixed with large-volume reagents, and the risk of loss of active ingredients due to pipetting wall hanging is also smaller. Multiple pumping filling units can efficiently fill multiple groups of large-volume reagents to be filled at the same time, and the filling unit configuration of five can complete the simultaneous and efficient filling of magnetic bead diluent, lysis solution, washing solution I, washing solution II, and washing solution III.

[0022] 3. At least some wells in the kit are lysis wells, into which the dispensing unit dispenses lysis buffer, and the pipetting assembly transfers activation buffer containing proteinase K and internal control. At least some wells in the kit are magnetic bead wells, into which the dispensing unit dispenses magnetic bead diluent, and the pipetting assembly transfers the vibrated magnetic bead mixture from the magnetic bead mixing assembly. This configuration eliminates the need for pre-mixing magnetic beads, complex high-precision pumping equipment, and the risk of magnetic bead aggregation and blockage due to intermittent dispensing. The kit's real-time preparation module also avoids the use of stirring equipment, preventing the inaccurate quantitative dispensing of magnetic bead solution by gravimetric dispensing. The eccentric oscillation scheme ensures good mixing and prevents damage to the magnetic bead coating due to excessive localized force. The amount of magnetic bead diluent dispensed is more than three times that of the magnetic bead solution, and the amount of lysis buffer dispensed is more than three times that of the activation buffer, accommodating the differences in precision and volume between the two different transfer components.

[0023] 4. The reagent preparation area of ​​the present invention includes a proteinase K carrying area, an internal control carrying area, and an activation solution preparation area, which centralizes reagent storage and preparation, reduces the travel distance of the pipetting assembly, and makes the reagent kit preparation more efficient. Attached Figure Description

[0024] Figure 1 This is a schematic diagram of the instant preparation module for the reagent kit of the present invention;

[0025] Figure 2 This is a schematic diagram of the real-time preparation module of the reagent kit of the present invention from another perspective;

[0026] Figure 3 This is a schematic diagram of the liquid circuit connection of the pumping and refueling assembly of the present invention;

[0027] Figure 4 This is a schematic diagram of the layout structure of the various components of the pumping and filling assembly for liquid connection in this invention;

[0028] Figure 5 This is a schematic diagram of the integrated layout structure of the reagent container of the present invention;

[0029] Figure 6 This is a schematic diagram of the centralized layout structure of the reagent container and various consumables of the present invention;

[0030] Figure 7 This is a schematic diagram of the combined layout of different filling pumps and valves of the present invention;

[0031] Figure 8 This is a schematic diagram of the structure of the operation layer of the present invention;

[0032] Figure 9 This is a layout diagram of the operation layer of the present invention from another perspective;

[0033] Figure 10A schematic diagram of the pumping and filling operation of the pumping and filling assembly of the present application to perform the pumping and filling operation of the reagent to be filled into the extraction reagent kit;

[0034] Figure 11 A state diagram of the transfer of the second type of reagent bottle by the tube clamping unit in the instant preparation module of the reagent kit of the present application;

[0035] Figure 12 A structure diagram of the preparation layer of the present application;

[0036] Figure 13 A structure diagram of the magnetic bead mixed liquid oscillation assembly of the present application;

[0037] Figure 14 A sectional view of the magnetic bead mixed liquid oscillation assembly of the present application;

[0038] Figure 15 A state diagram of the transfer of the reagent to be transferred by the first pipetting unit of the present application;

[0039] Figure 16 A state diagram of the loading of the magnetic rod sleeve into the reagent kit by the extraction magnetic rod sleeve loading unit in the instant preparation module of the reagent kit of the present application;

[0040] Figure 17 A state diagram of the transfer of various consumables by the consumable clamping and transfer unit in the instant preparation module of the reagent kit of the present application.

[0041] Marked in the figure: 10-fixed cross beam, 101-fixed cross beam drive motor, 102-fixed cross beam drive wheel, 103-fixed cross beam transmission belt, 105-fixed beam connecting block, 104-fixed beam slide rail, 106-fixed cross beam driven wheel, 110-pumping filling assembly, 1101-filling unit, 111-liquid pressure sensor, 122-filling lifting belt, 123-filling lifting transmission wheel, 21-extraction preparation position, 211-preparation drive motor, 212-preparation drive wheel, 213-preparation transmission belt, 214-preparation connecting block, 215-preparation slide rail, 22-washing tank, 23-magnetic bead mixed liquid oscillation assembly, 231-oscillation receiving position, 232-oscillation table, 233-oscillation base, 234-oscillation damping element, 2351-oscillation motor, 2352-oscillation eccentric wheel, 24-transport channel, 25-reagent loading drawer, 251-first type reagent bottle storage position, 252-second type reagent bottle storage position, 253-reagent bottle recycling position, 26-reagent dispensing area, 261, 262, 263-reagent bottle receiving position, 264-bottle cap carrying table, 265-fixing unit, 266-reagent bottle detection unit, 271-first consumable temporary storage position, 272-second consumable temporary storage position, 273-magnetic rod sleeve temporary storage position, 31-fluid driving module, 311-filling pump, 312-first three-way valve, 313-second three-way valve, 314-washing two-way valve, 315-washing three-way valve, 316-washing pump, 317-waste liquid recovery pump, 319-waste liquid three-way valve, 32-reagent barrel carrying part, 320-waste liquid barrel, 321-washing liquid barrel, 322-reagent barrel to be filled, 323-bottle cap unit, 324-reagent needle, 325-bottle cap buffer, 33-barreled reagent drawer, 331-reagent drawer drive motor, 332-reagent barrel receiving position, 34-consumable storage part, 341-consumable loading position, 3411-consumable lifting drive wheel, 3412-consumable lifting belt, 3413-consumable lifting driven wheel, 3414-consumable lifting slide rail, 3415-consumable lifting connecting block, 351-first consumable area, 352-second consumable area, 40-first movement cross beam, 401-first movement drive motor, 402-first drive wheel, 403-first transmission belt, 404-first slide rail, 405-first connecting block, 410-first pipetting unit, 420-tube clamping unit, 50-second movement cross beam, 501-second movement drive motor, 502-second drive wheel, 503-second transmission belt, 504-second slide rail, 505-second connecting block, 510-magnetic rod sleeve loading unit, 5101-magnetic rod sleeve intake unit, 511-magnetic rod sleeve loading lifting drive motor, 512-magnetic rod sleeve connecting block, 520-consumable clamping and transferring unit. DETAILED DESCRIPTION

[0042] The application will be described in detail below with reference to the drawings.

[0043] In order to make the objectives, technical solutions and advantages of the present application clearer, the present application will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present application and not intended to limit the present application.

[0044] Embodiment 1

[0045] As described in the background, most of the integrated large-scale in vitro diagnostic devices developed by the head enterprises are designed to configure various types of reagent kits required for on-site detection. The molecular diagnostic technology taking genetic material nucleic acid fragments as the detection object first needs to obtain the nucleic acid fragments to be detected from various types of samples. This processing process is called extraction and purification process. Generally speaking, it contains multiple independent steps. In some specific extraction steps, multiple reagents may be required. The current solution is to mix and configure before use and then add. However, this method has the problem of inaccurate mixing ratio and puts forward higher requirements for the operator.

[0046] Figure 1 And Figure 2Respectively, the structural diagram of different perspectives of the kit instant preparation module; the kit instant preparation module can be spliced by connecting beams, various types of substrates and other structural members; in order to ensure sufficient space for operation, the kit instant preparation module is configured in the shape of a near cuboid structure, and the overall layout of the kit instant preparation module can be classified into three different processing layers from top to bottom; the uppermost layer can be configured as an operation layer, which can drive clamping units, pipetting units and other functional units to perform different operation functions such as transfer, pipetting, filling and the like; the middle layer is a preparation layer composed of various types of storage site operation sites; and the lowermost layer is a basic functional layer for centralized addition of various types of reagents and consumables; the various functional units of the operation layer are configured on two movable motion cross beams and a fixed cross beam 10 which can slide longitudinally along the kit instant preparation module, where the longitudinal direction refers to the X-axis direction; the movable motion cross beams include a first movable motion cross beam 40 and a second movable motion cross beam 50 which are independent of each other and can be driven to move between various operation sites of the preparation layer; the preparation layer includes a reagent loading drawer 25 which can be driven to extend out of the main body range of the kit instant preparation module to transport various types of reagents; in order to ensure the running stroke of the reagent loading drawer 25, the transmission mechanism thereof is configured as a folding transmission structure; the reagent loading drawer 25 includes a first type of reagent bottle receiving site 251, a second type of reagent bottle receiving site 252 and a reagent bottle recycling site 253 on the substrate thereof; the first type of reagent bottle receiving site includes a plurality of arrayed first type of reagent bottle receiving holes for receiving proteinase K reagent bottles, internal reference reagent bottles and dispensing bottles in groups; this example shows that a total of 16 first type of reagent bottle receiving holes are included, of which 2*4 are proteinase K reagent bottle receiving holes, 1*4 are internal reference reagent bottle receiving holes and 1*4 are dispensing bottle receiving holes; the specific number is not limited here; the second type of reagent bottle receiving site 252 can include two independent second type of reagent bottle receiving holes for receiving magnetic bead mixing reagent bottles and eluent reagent bottles; the magnetic bead mixing reagent bottles and the eluent reagent bottles can have basically the same appearance and can be distinguished by color and other distinguishing features of the reagent bottle caps to enable the kit instant preparation module to automatically recognize them, thereby enabling the operator to add different types of reagents at will; the basic functional layer includes a consumable carrying drawer which includes various types of consumable loading sites 341; the number of consumable loading sites 341 is not limited and can be multiple; they can carry eight-pipe amplification consumables, pipetting consumables, deep-well plate extraction consumables and extraction magnetic rod sleeves and other types of consumables; centralized addition of various types of consumables can facilitate operator operation and also help to plan the flow path of various types of consumables to make the entire flow process more convenient; the consumable loading part can be divided into a first consumable area 351 and a second consumable area 352 which can be independently driven to extend out of or retract into the main body range of the kit instant preparation module by independent drive motors;Of course, in order to make the kit instant preparation module have higher integration, a reagent barrel bearing part 32 is arranged at the lower part of the reagent loading drawer 25, which contains a plurality of reagent barrel receiving positions 332, each of which can be arranged with a weight sensor. The reagent barrel bearing part 32 is assembled in the barrel reagent drawer 33, and the reagent barrel receiving positions 332 of the reagent barrel bearing part 32 can be driven by the barrel reagent drawer 33 to be pulled out or brought back to the kit instant preparation module, thereby receiving the reagent barrel to cooperate with the liquid filling operation. Adjacent to the reagent loading drawer 25, a transfer channel 24 and a reagent preparation area 26 are arranged in the longitudinal direction, where the longitudinal direction refers to the X-axis direction. The transfer channel 24 is arranged with a rotatable conveyor belt, which can transfer different types of consumables transferred thereon to other kit instant preparation modules in cooperation with the consumable transfer clamping unit. The reagent preparation area 26 includes a proteinase K reagent bottle bearing area, an internal reference reagent bottle bearing area, and an activation liquid preparation area. In addition, a reagent bottle cap receiving position is arranged at one end of the reagent preparation area 26 to independently receive the reagent bottle cap after opening different types of reagent bottles. The magnetic bead mixed liquid oscillation assembly 23 is connected to the bottom substrate of the kit instant preparation module through a vibration absorption and damping element, which can maximize the absorption of vibration to ensure the reliable operation of the kit instant preparation module. The assembly can mix the bottled magnetic bead mixed liquid more uniformly and efficiently. Since the magnetic bead mixed liquid uses a pipetting assembly for high-precision transfer, it does not need to configure a gravity sensor similar to a pumping filling to control the filling amount. Adjacent to the consumable bearing drawer, a first consumable temporary storage position 271 and a second consumable temporary storage position 272 are arranged. The first consumable temporary storage position 271 and the second consumable temporary storage position 272 can accommodate different types of consumables, such as extraction magnetic rod sleeve boxes and pipetting consumables.The fixed cross beam 10 is provided with a fixed cross beam driving motor 101, and a fixed cross beam driving wheel 102 and a fixed cross beam driven wheel 106 are arranged in the length direction (i.e. the Y-axis direction) of the fixed cross beam 10. The fixed cross beam driving wheel 102 and the fixed cross beam driven wheel 106 are spaced apart by a predetermined distance, and a fixed cross beam transmission belt 103 is arranged between the fixed cross beam driving wheel 102 and the fixed cross beam driven wheel 106. One side of the fixed cross beam transmission belt 103 is fixedly connected with a fixed beam connecting block 105, and the fixed beam connecting block 105 is matched with a fixed beam sliding rail 104. A pumping and filling assembly 110 is connected to the fixed beam connecting block 105. The fixed cross beam driving wheel 102 can be driven to rotate by the fixed cross beam driving motor 101, and in turn drive the fixed beam connecting block 105 to slide along the fixed beam sliding rail 104 through the fixed cross beam transmission belt 103, so that the pumping and filling assembly 110 moves along the extension direction of the fixed beam sliding rail 104. The pumping and filling assembly 110 can also be driven to move up and down by a pumping and lifting driving mechanism. An extraction preparation site 21 matched with the pumping and filling assembly 110 on the fixed cross beam 10 is further arranged on the preparation layer. The extraction preparation site 21 is used for instant configuration of the extraction reaction reagent box. The driving device of the extraction preparation site 21 includes a preparation driving motor 211, and the output shaft of the preparation driving motor 211 is connected with a preparation driving wheel 212. A preparation driven wheel is arranged at a predetermined distance along the longitudinal direction of the reagent box instant preparation module. The longitudinal direction here refers to the X-axis direction. A preparation transmission belt 213 is arranged between the preparation driving wheel 212 and the preparation driven wheel. The extraction preparation site 21 is connected to the preparation transmission belt 213 through a preparation connecting block 214, and the preparation connecting block 214 is matched with a preparation sliding rail 215. The preparation driving motor 211 drives the preparation driving wheel 212 to rotate, so that the preparation connecting block 214 moves along the preparation sliding rail 215 under the action of the preparation transmission belt 213, and in turn moves the extraction preparation site 21 along the extension direction of the preparation sliding rail 215, so as to cooperate with the pumping and filling assembly 110 to complete the filling of the to-be-filled liquid or cooperate with the pipetting assembly to complete the transfer of the to-be-transferred reagent, and finally complete the instant configuration of the extraction reaction reagent box.

[0047] Figure 3Fig. 2 is a schematic diagram of a liquid connection of the pumping and filling assembly 110 of the application; the inventor found that the amount of different types of reagents required in different operation steps is different, and the reagent with a larger liquid requirement needs to be filled by pumping method, which can adapt to the demand of continuous multiple filling, and also does not need to plan too much reagent storage area, and the reagent barrel or reagent tank in the form of several liters, several tens of liters or even several hundred liters can directly complete the batch addition of the reagent to be filled. In this embodiment, the lysis solution, washing I, washing II, washing III and magnetic bead diluent in the extraction step can be filled into different reagent barrels 322 to be filled, and the first three-way valve 312 and the second three-way valve 313 are respectively in fluid communication with the corresponding same number of filling pumps 311, and the filling pumps 311 are also in fluid communication with the plurality of filling units 1101 of the pumping and filling assembly 110 through the second three-way valve 313, and a liquid pressure sensor 111 is arranged in the fluid communication pipeline upstream of the filling unit 1101, so that whether the filling operation of the different filling units 1101 is normal can be monitored, and it can also be used as a control signal for the operation of the pumping and filling assembly 110, and the first three-way valve 312 is also in fluid communication with the cleaning liquid barrel 321 through a plurality of branch pipelines, and the reagent kit instant preparation module is also provided with a cleaning tank 22, which is in fluid communication with the cleaning liquid barrel 321 and the cleaning pump 316 by using the cleaning two-way valve 314 and the cleaning three-way valve 315, and the cleaning tank 22 is provided with a cleaning hole and an overflow groove, and the cleaning hole and the overflow groove are in fluid communication with the waste liquid barrel 320 through the waste liquid three-way valve 319 and the waste liquid recovery pump 317. When it is needed to fill various reagents into the extraction reaction reagent kit in the extraction preparation site 21, the plurality of filling units 1101 are in fluid communication with various reagent barrels 322 to be filled, so that they can pump and fill a predetermined volume of reagent to be filled into at least part of the hole site of the extraction reaction reagent kit; here the volume of the reagent to be filled can be in the order of hundreds of microliters, for example, the filling amount can be 300 μL, 350 μL, 400 μL, 450 μL, 500 μL, 550 μL, etc., so that the pumping and filling assembly 110 only needs to ensure that the appropriate amount of various extraction reagents is filled into at least part of the hole site of the extraction reaction reagent kit through the pumping time and / or the weight signal of the gravity sensor at the reagent barrel receiving site 332; and when the predetermined time filling operation or the pressure detection value of the filling system is abnormal, the pumping and filling assembly 110 can be driven to move above the cleaning tank 22 to perform cleaning operation, and in the cleaning step, the cleaning liquid in the cleaning liquid barrel 321 can be driven into the cleaning tank 22 through a plurality of filling pumps 311 and a cleaning pump 316 through different flow channels, thereby realizing simultaneous cleaning of the inner and outer walls of the pumping and filling assembly 110, so that the system can always efficiently and accurately pump various types of reagents to be filled.In the embodiment, the five reagent buckets 322 to be filled are provided to fill five kinds of reagents at the same time, and the corresponding number of filling pumps 311, filling units 1101, first three-way valves 312 and second three-way valves 313 is five, which can realize faster and more accurate configuration of the extraction system. In some special cases, other numbers can also be configured, which are not limited here.

[0048] Figure 4 Figure is a schematic view of the layout of various components of the liquid connection of the pumping and filling assembly 110 of the application; the pumping and filling assembly 110 is arranged on the fixed cross beam 10 and can be driven to slide along the length direction of the fixed cross beam 10 by the fixed cross beam driving motor 101. The fixed cross beam 10 is arranged in the transverse direction, which is the Y-axis direction. The extraction preparation position 21 and the cleaning tank 22 are arranged in parallel in the transverse direction below the fixed cross beam 10. When the filling unit 1101 is driven to align with one of them, the filling lifting driving motor can drive the filling unit 1101 to lift through the filling lifting driving wheel, the filling lifting transmission wheel 123 and the filling lifting belt 122, thereby completing the filling or cleaning operation. The extraction preparation position 21 can be driven by the preparation driving motor 211 through the pulley transmission mechanism, thereby driving the preparation connecting block 214 to slide along the reagent cartridge instant preparation module in the longitudinal direction, which is the X-axis direction. In this way, on the one hand, the extraction reaction reagent cartridge holes in the extraction preparation position 21 can be aligned with the filling unit 1101, and the reagent filling in all the holes of the extraction reaction reagent cartridge can be completed under the premise that the filling unit 1101 is stationary. The matching area of the extraction preparation position 21 and the filling unit 1101 forms a filling position. On the other hand, the extraction preparation position 21 can be moved to a pipetting position which is spaced apart from the filling position by a predetermined distance, thereby cooperating with the pipetting assembly to perform pipetting addition of another part of the reagent. The pumping and filling assembly 110 in the reagent cartridge instant preparation module is arranged on the fixed cross beam 10 to realize simplified arrangement of the pipeline of the liquid system, thereby making the reagent cartridge instant preparation module more reliable and reducing the risk of tearing, damage and breakage of the fluid communication pipeline. The fluid driving module 31 is also arranged on the side of the reagent cartridge instant preparation module, and the fluid driving module 31 includes the filling pump 311, the first three-way valve 312, the second three-way valve 313, the cleaning two-way valve 314, the cleaning three-way valve 315, the cleaning pump 316, the waste liquid recovery pump 317 and the waste liquid three-way valve 319. The reagent bucket bearing part 32 is also slidably assembled on the bottom substrate.

[0049] Figure 5is a schematic diagram of the integrated layout structure of the reagent bucket of the application; the reagent bucket bearing part 32 comprises a plurality of reagent bucket receiving positions 332, each of which comprises a gravity sensing unit; the reagent bucket receiving positions 332 are arranged in the reagent bucket loading drawer 33 as a whole and can be driven to extend or retract within the main range of the preparation layer by the reagent drawer driving motor 331; the reagent buckets carried by the reagent bucket receiving positions 332 can comprise a waste liquid bucket 320, a cleaning liquid bucket 321 and a plurality of reagent buckets to be filled 322; the positions of different reagent buckets are not limited here; above the corresponding reagent buckets, a bottle cap unit 323 capable of lifting motion is arranged, and a reagent needle 324 in fluid communication with the liquid path of the pumping and filling assembly 110 is arranged, which can be driven to lift and is arranged near the reagent bucket opening and is provided with a bottle cap buffer 325, which can buffer and support the descending bottle cap unit 323 to make the filling operation more reliable.

[0050] Figure 6 is a schematic diagram of the centralized layout structure of the reagent bucket and various consumables of the application; the reagent bucket bearing part 32 and the consumable storage part 34 are arranged side by side, which can realize the centralized addition of various consumables and bucket reagents, and the operator can also check whether the addition of different consumables is normal more conveniently, and the replacement operation is also more simple and convenient; here, the consumable storage part 34 comprises a plurality of storage positions for storing pipette consumables, extraction magnet sleeve, deep well plate consumables, amplification eight-tube consumables and the like; the number of storage positions is not limited, all storage positions can be divided into two independently driven consumable bearing drawers to facilitate the addition of consumables; of course, all storage positions can also be arranged on the same consumable bearing drawer, so that the drawer driving is more simple and convenient; a consumable lifting mechanism is arranged on each consumable bearing drawer, which comprises a consumable lifting driving motor; the output shaft of the consumable lifting driving motor is connected with a consumable lifting driving wheel 3411; a consumable lifting driven wheel 3413 is arranged at a preset distance from the consumable lifting driving wheel 3411 in vertical height; a consumable lifting belt 3412 is arranged between the consumable lifting driving wheel 3411 and the consumable lifting driven wheel 3413; a consumable lifting connecting block 3415 is fixedly connected to the consumable lifting belt 3412; the consumable lifting connecting block 3415 can be driven to slide along the vertically arranged consumable lifting slide rail 3414 and drive the consumable bearing disc in the consumable loading position 341 to move upward, so that the various stacked consumables can be transferred by the consumable clamping unit on the top of the consumable loading position 341.

[0051] Figure 7 is a schematic diagram of the combined layout of different filling pumps 311 and valve elements of the application; here, the corresponding liquid path system Figure 3The number of reagent barrels 322 to be filled in the structure is five. In order to ensure the compactness of the structure and the configuration requirements of the reagent box instant preparation module, the filling pump 311 is divided into two groups and configured on two substrates. Five first three-way valves 312 and five second three-way valves 313 are respectively configured on the substrates of the two groups of filling pumps 311. The cleaning pump 316 is configured on a different substrate from the filling pump 311. The cleaning pump 316 is further configured with a cleaning two-way valve 314, a cleaning three-way valve 315, and a waste liquid three-way valve 319 on the substrate. The waste liquid recovery pump 317 is arranged separately. Each component is connected according to the liquid path system diagram using pipelines, three-way valves, two-way valves, elbows, joints, and the like. The filling pump 311 and the cleaning pump 316 can be the same type of liquid pump, for example, an injection pump. The waste liquid recovery pump 317 is a different type of liquid pump, for example, a diaphragm pump. According to such modular arrangement, the mutual influence of each pump and valve during operation can be reduced, and the system operation reliability is higher.

[0052] Figure 8 and Figure 9Respectively are structural schematic diagram of different visual angle of operation layer of the application; The uppermost operation layer contains fixed cross beam 10, which is arranged in the vertical direction of the instant preparation module of the kit away from the sample adding unit such as reagent drawer, the vertical direction here refers to the direction along the Z axis, a pumping filling assembly 110 is slidably assembled thereon and can slide along the length direction of the fixed cross beam 10, the movement drive has been described before and will not be repeated here. Two independently movable movement cross beams, i.e. the first movement cross beam 40 and the second movement cross beam 50, are also arranged at the position vertically higher than the fixed cross beam 10, the two movement cross beams can be driven by two movement cross beam drive motors through independent movement cross beam transmission mechanisms to slide along the common slide rail arranged in the longitudinal direction of the instant preparation module of the kit, the longitudinal direction here refers to the direction along the X axis, in order to ensure low resistance and reliability of movement, the other end of the two movement cross beams is in rolling contact with the common guide rod through a roller set. The first movement drive motor 401 is arranged on the first movement cross beam 40, the output shaft of the first movement drive motor 401 is connected with the first drive wheel 402, the first driven wheel is arranged at a preset interval in the length direction of the first movement cross beam 40, the first transmission belt 403 is arranged between the first drive wheel 402 and the first driven wheel, the first connecting block 405 is arranged on the first transmission belt 403, when the first movement drive motor 401 rotates, the first connecting block 405 can slide along the first slide rail 404 through transmission, so as to drive the functional units on the first movement cross beam 40 to move in the length direction of the first movement cross beam 40, the width direction of the first movement cross beam 40 contains the first pipetting unit 410 and the tube clamping unit 420 on both sides, the width direction here refers to the direction along the X axis, in order to improve the preparation efficiency of the reaction reagent kit in the instant preparation module of the kit, the second pipetting unit is also contained on the arrangement side of the tube clamping unit 420, the first pipetting unit 410 and the second pipetting unit can be connected with pipetting consumables of different capacities to transfer different types and different volumes of reagents, i.e. the first pipetting unit 410 and the second pipetting unit can jointly constitute a pipetting assembly. The tube clamping unit 420 can not only perform clamping and transfer of various reagent tubes, but also can perform cap opening and closing operation of the reagent tube in cooperation with other fixed units. The second movement drive motor 501 is arranged on the second movement cross beam 50, the output shaft of the second movement drive motor 501 is connected with the second drive wheel 502, the second driven wheel is arranged at a preset interval in the length direction of the second movement cross beam 50, the length direction here refers to the direction along the Y axis, the second transmission belt 503 is arranged between the second drive wheel 502 and the second driven wheel, the second connecting block 505 is arranged on the second transmission belt 503, when the second movement drive motor 501 rotates, the second connecting block 505 can slide along the second slide rail 504 through transmission, so as to drive the functional units on the second movement cross beam 50 to move in the length direction of the second movement cross beam 50, the width direction of the second movement cross beam 50 contains the extraction magnet bar sleeve loading unit 510 and the consumable clamping and transfer unit 520 on both sides.

[0053] Figure 10 Figure 13 is a schematic diagram of the pumping and filling assembly 110 of the present application performing the filling operation of the liquid to be filled in the extraction reaction kit; after the various reagents and consumables in the instant preparation module of the kit are correctly added, the extraction preparation position 21 can be driven to run by the preparation driving motor 211 to a position where at least part of the hole position is aligned with the pumping and filling assembly 110, at this time the pumping and filling assembly 110 on the fixed cross beam 10 can be driven to descend so that at least part of the filling unit 1101 is immersed in part of the hole position of the extraction reaction kit, and then the filling pump 311 performs the preset amount of liquid filling according to the filling time, weight change and / or pressure change, etc. In this embodiment, the instant preparation module of the kit first performs pumping and filling of a larger volume of liquid to be filled in the extraction reaction kit, and the filling amount can be pre-configured in the control module, so that a certain liquid level height of the base liquid can be formed in the extraction reaction kit, and then a smaller volume of reagent to be transferred is transferred by using the first pipetting unit 410 or the second pipetting unit, so that the loss of the reagent to be transferred caused by wall hanging can be minimized, and better mixing effect can be achieved, and problems such as splashing loss caused by pumping and filling of the reagent to be filled into the reagent added by pipetting can also be eliminated. The extraction reaction kit after filling can be driven to move to the pipetting receiving position by the preparation layer, and further complete the addition of all reagents under the cooperation of the pipetting assembly.

[0054] Figure 11 Figure 14 is a structural diagram of the preparation layer of the present application; when the first type of reagent bottle storage position 251 and the second type of reagent bottle storage position 252 are placed with reagent bottles of various types, the tubular clamping unit 420 can be driven by the first movement cross beam 40 to transfer different reagent bottles from the reagent loading drawer 25 to the various functional areas of the reagent dispensing area 26, the functional areas of the reagent dispensing area 26 include a proteinase K carrying area, an internal reference carrying area and an activation liquid dispensing area, each functional area is provided with a corresponding reagent bottle receiving position, in Figure 11 Figure 15 is a schematic diagram of the reagent bottle receiving position in Figure 14; the reagent bottle receiving position in Figure 14 has three reagent bottle receiving positions 261, 262 and 263, which are respectively a proteinase K receiving position, an internal reference receiving position and a dispensing receiving position, each reagent bottle receiving position contains a fixing unit 265, which can clasp and fix various reagent bottles transferred by the tubular clamping unit 420 in different receiving positions, in order to ensure that each reagent bottle receiving position can accurately determine whether the reagent bottle is correctly received therein, a reagent bottle detection unit 266 is also configured in each reagent bottle receiving position, the reagent bottle after transfer can be opened by the tubular clamping unit 420, and the opened reagent bottle cap is placed at different positions of the bottle cap carrying table 264.

[0055] Figure 12is the state diagram of the transfer of the second type of reagent bottle by the tube clamping unit 420 in the instant kit preparation module of the present application; the tube clamping unit 420 can transfer the eluent reagent bottle in the second type of reagent bottle to the eluent pipetting position by clamping, and the inventors notice that the amount of eluent used in the extraction process is smaller than that of other reagents, and the order of magnitude is in the order of tens of microliters or hundreds of microliters, so here the eluent is also transferred by the pipetting assembly, which can ensure the accuracy of the transfer amount on the one hand, and also reduce the number of cleaning of the filling unit 1101, increase the reliability of the filling unit 1101 (mainly because more filling units 1101 means higher risk of blockage and higher frequency of cleaning operation), the tube clamping unit 420 can transfer the eluent reagent bottle to the eluent pipetting position, and perform the uncapping of the eluent reagent bottle, and then after the pumping and filling are completed, a set amount of eluent can be transferred to the elution hole position of the extraction reaction reagent kit by pipetting.

[0056] Figure 13 is a structure diagram of a magnetic bead mixed liquid oscillation assembly of the present application, Figure 14is a sectional view of the magnetic bead mixed solution oscillation assembly of the present application; the tubular clamping unit 420 can transfer the magnetic bead mixed solution reagent bottle to the magnetic bead mixed solution oscillation assembly 23 by clamping the bottle cap of the second type of reagent bottle, the magnetic bead mixed solution in the present application can be directly obtained from the mature commercial magnetic bead mixed solution bottle added from the loading part, and the magnetic bead mixed solution always exists in a concentrated state, so its storage time is also longer and the performance is more stable, the operator does not have to mix the diluent with it in advance, because the volume of the concentrated magnetic bead mixed solution is smaller, the energy consumption required for mixing is also smaller, and it is not necessary to use a stirring type mixing scheme similar to the prior art, because the transfer volume of the concentrated magnetic bead mixed solution is smaller and meets the requirements of the accuracy range of the pipette, the transfer volume of the magnetic bead mixed solution is generally not more than 80 μL, compared with the larger transfer volume (not less than 3 times the pipetting volume) after mixing with the magnetic bead diluent, it is difficult to use the pipette to transfer, and in the pumping and filling scheme, the performance requirements of the filling pump 311 of the solid-liquid two-phase medium are higher, and the interval of the filling operation will cause the risk of magnetic bead agglomeration and blockage in the system, resulting in lower system operation reliability, and it is difficult to be accurate all the time. Because stirring is required, the accuracy of the filling amount controlled by the weighing method is lower. The uppermost part of the magnetic bead mixed solution oscillation assembly 23 is the oscillation receiving site 231, the top open end of which can receive the magnetic bead mixed solution reagent bottle transferred, and the bottom is connected to the oscillation table 232, the oscillation table 232 is also connected to the oscillation base 233, the oscillation base 233 is connected to the fixed rod and the vibration absorption base through a plurality of vibration absorption elements 234, so that the vibration of the oscillation receiving site 231 has little effect on other components in the reagent box immediate preparation module. The oscillation motor 2351 and the oscillation eccentric wheel 2352 are respectively arranged on the oscillation table 232 and the oscillation base 233, the rotation movement output by the oscillation motor 2351 drives the eccentric rotation movement of the oscillation eccentric wheel 2352, and then the magnetic bead mixed solution reagent bottle on the oscillation receiving site 231 is shaken and oscillated, and then the magnetic bead mixed solution is always subjected to vortex effect to mix the magnetic beads. Only when pipetting transfer is required, the rotation is stopped to ensure that the magnetic bead mixed solution is always in a more uniform state, and the pipetting accuracy of the pipetting assembly is much higher than that of the pumping and filling scheme, so that the reagent filling of the pumped magnetic bead diluent and the pipetted magnetic bead mixed solution is more stable (the filling amount of the magnetic bead diluent can be more than 3 times the filling amount of the magnetic bead mixed solution).

[0057] Figure 15is a schematic diagram of the first pipetting unit 410 of the present application executing the transfer state of the reagent to be pipetted; in the present embodiment, the inventors have noticed that although premixing pump filling has a certain convenience, it has other reliability problems such as salting out blockage and corrosion; here, the two reagents of protease K and internal reference have a small filling amount, and generally the filling amount of the two reagents in one extraction hole is 10 μL, 15 μL, 20 μL, 25 μL, 30 μL, 35 μL, 40 μL, 45 μL, 50 μL, etc. The use of the first pipetting unit 410 or the second pipetting unit can more accurately control the addition amount. The pipetting assembly can first aspirate a preset volume of protease K from the protease K reagent bottle and pipette it into the preparation bottle, and then aspirate a preset volume of internal reference reagent from the internal reference reagent bottle and transfer it to the preparation bottle. The pipetting assembly can mix the two reagents in the preparation bottle by multiple suction operations. The pipetting sequence of the two reagents is not limited. Finally, the pipetting assembly can pipette the mixed activation solution into the filling hole of the lysis solution. Of course, in order to adapt to different pipetting amounts and pipetting precision requirements, the pipetting assembly is configured as the first pipetting unit 410 and the second pipetting unit. The first pipetting unit 410 and the second pipetting unit are connected with pipetting consumables of different capacities to efficiently and accurately execute the transfer of different reagents.

[0058] Figure 16 is a state diagram of the extraction magnet rod sleeve loading unit in the instant preparation module of the kit of the present application loading the magnet rod sleeve into the extraction reaction kit after completing the transfer of the reagent; all the holes of the extraction reaction kit after filling and pipetting have been correspondingly configured with various types of extraction reagents. At this time, the extraction preparation site 21 can still stay at the pipetting site. The second movement cross beam 50 can drive the magnet rod sleeve loading unit 510 to move to the upper side of the magnet rod sleeve temporary storage site 273. Then the magnet rod sleeve loading lifting drive motor 511 can drive the magnet rod sleeve drive screw 512 and the magnet rod sleeve connecting block 513 connected by screwing to make the magnet rod sleeve loading unit 510 descend and connect a corresponding number of magnet rod sleeves through the plurality of magnet rod sleeve suction units 5101. Then it rises and moves to the upper side of the extraction reaction kit in the pipetting site with the second movement cross beam 50. Here, the plurality of magnet rod sleeve suction units 5101 can be aligned with the magnetic bead storage site of the kit. Then it is driven downward by the magnet rod sleeve loading lifting drive motor 511 and finally loaded into the magnetic bead storage hole of the extraction reaction kit with the assistance of the unloading unit, completing the instant configuration of the extraction reaction kit.

[0059] Figure 17The consumable clamping and transferring unit 520 of the instant kit preparation module of the present application transfers various consumable state diagrams; the extraction reaction kit with loaded magnetic rod sleeve can be clamped by the consumable clamping and transferring unit 520 on the second moving cross beam 50 and transferred to the transmission channel 24, so that the instant configuration extraction reaction kit can be used for nucleic acid extraction operation in other subsequent related modules. Of course, the consumable clamping and transferring unit 520 can also transfer a new empty extraction reaction kit to the extraction preparation site 21, can supplement the transfer of the extraction magnetic rod sleeve box to the magnetic rod sleeve temporary storage site 273, can transfer the first pipette consumable box to the first consumable temporary storage site 271, can transfer the second pipette consumable box to the second consumable temporary storage site 272, and can also transfer various boxes from the corresponding positions to the transmission channel 24. Of course, the eight-pipe amplification consumable box can also be transferred to the transmission channel 24. Here, there is no limitation.

[0060] Therefore, the present application also discloses a sample processing system comprising a kit instant preparation module, and more preferably, the sample processing system is a flow sample processing system, comprising a sample tube storage and flow module, a kit instant preparation module, an extraction processing module, an amplification system preparation module and an amplification detection module, so that large-scale continuous sample processing can be realized under the premise of high-integration layout.

[0061] The kit instant preparation method of the kit instant preparation module of the present application also discloses a kit instant preparation method, especially an extraction reaction kit preparation method, comprising a pumping and filling assembly 110, the pumping and filling assembly 110 comprising not less than one filling unit 1101, the filling unit 1101 being in fluid communication with a to-be-filled reagent barrel 322 through a filling pump, and the filling unit 1101 being capable of pumping and filling a preset volume of to-be-filled reagent into at least part of the holes of the extraction reaction kit; and a pipetting assembly, the pipetting assembly being capable of sucking a preset volume of to-be-pipetted reagent from a to-be-pipetted reagent bottle and transferring it to at least part of the holes of the kit, so that all the holes receiving the to-be-pipetted reagent in the kit also contain to-be-filled reagent with a volume greater than that of the to-be-pipetted reagent. The inventor classifies different reagents of the extraction reaction kit, does not need to premix before machine operation, maintains the original storage state of various reagents, and then utilizes the automatic pumping and filling assembly 110 and the pipetting assembly to transfer different amounts of reagents from various reagent barrels or reagent bottles to mix in the holes to form the instant-prepared extraction reaction kit, which can ensure accurate filling amount of various reagents of the extraction reaction kit and higher extraction efficiency.

[0062] The adding unit 1101 can first pump a preset volume of to-be-added reagent into at least part of the wells of the extraction reaction kit, and then the pipetting assembly can transfer a preset volume of to-be-pipetted reagent into at least part of the wells of the extraction reaction kit that have been added with the to-be-added reagent. The adding unit 1101 is in a plurality of quantity, and the plurality of adding units 1101 are in fluid communication with a corresponding number of to-be-added reagent barrels 322 through a plurality of adding pumps in the same number. The adding unit 1101 includes a magnetic bead diluent adding part and a lysis solution adding part. The pipetting assembly can pipette and transfer a first volume of magnetic bead mixed solution into at least part of the wells of the extraction reaction kit, and all the wells receiving the magnetic bead mixed solution further contain a second volume of magnetic bead diluent pumped and added by the magnetic bead diluent adding part. The pipetting assembly can pipette and transfer a third volume of activation solution containing proteinase K and an internal reference into at least part of the remaining wells of the extraction reaction kit, and all the wells receiving the activation solution further contain a fourth volume of lysis solution pumped and added by the lysis solution adding part. The second volume of magnetic bead diluent is not less than 3 times the volume of the first volume of magnetic bead mixed solution, and the fourth volume of lysis solution is not less than 3 times the volume of the third volume of activation solution. In this way, the difference in optimal precision range of different transfer operations and the requirement for long-time reliable operation can be adapted.

Claims

1. A kit instant chemistry preparation module, characterized by, The pumping filling assembly can pump and fill a preset volume of reagent to be filled into a designated well site, and the pipetting assembly can transfer a preset volume of pipetting reagent to the designated well site, and the well site to receive the pipetting reagent contains a volume of reagent to be filled that is greater than the pipetting reagent; the pumping filling assembly first fills a preset volume of each type of reagent into at least part of the wells of the reagent box, and then uses the pipetting assembly to pipette and transfer another type of reagent into at least part of the wells of the filled reagent box, so that a small volume of pipetting reagent is uniformly mixed in a large volume of reagent to be filled; the large volume of reagent to be filled includes a second volume of magnetic bead diluent, and the small volume of pipetting reagent includes a first volume of magnetic bead mixture, the magnetic bead mixture is in a concentrated state, and the second volume is not less than three times the first volume.

2. The kit-instant preparation module of claim 1, wherein, The pumping filling assembly includes a plurality of filling units, and the plurality of filling units are in fluid communication with corresponding reagent barrels through corresponding filling pumps.

3. The kit-instant preparation module of claim 1, wherein, It also includes a reagent preparation area, which contains a proteinase K carrying area, a reference carrying area, and an activation solution preparation area. The pipetting assembly can move between the proteinase K carrying area, the reference carrying area, and the activation solution preparation area, and transfer proteinase K and reference to the activation solution preparation area in a set volume ratio. The pipetting assembly can repeatedly perform liquid suction and discharge operations in the activation solution preparation area to obtain the activation solution by stirring and mixing.

4. The kit-instant preparation module of claim 1, wherein, It also includes a magnetic bead mixture oscillation assembly, which carries a magnetic bead mixture reagent bottle. The magnetic bead mixture oscillation assembly can oscillate and shake the magnetic bead mixture reagent bottle to generate a vortex effect to mix the magnetic beads in the magnetic bead mixture reagent bottle.

5. The kit-instant preparation module of claim 1, wherein, The pumping filling assembly is selectively in fluid communication with the washing liquid barrel or the reagent barrel through a liquid path assembly; it also includes a washing tank, which is in fluid communication with the washing liquid barrel through a washing pump; the pumping filling assembly can be moved to the washing tank for outer wall cleaning and can be connected to the washing liquid barrel for inner wall cleaning.

6. The kit-instant preparation module of claim 5, wherein, The pumping filling assembly is arranged on a fixed cross beam and can be driven by a fixed cross beam drive motor to slide along the length direction of the fixed cross beam. An extraction preparation site and a washing tank are arranged in parallel below the fixed cross beam. The pumping filling assembly can be driven to align with the extraction preparation site to complete the filling operation or to align with the washing tank to complete the cleaning operation. The extraction preparation site is equipped with an extraction reaction reagent box, which is provided with a plurality of wells arranged in an array in the horizontal and vertical directions. The extraction preparation site can be driven to move in the vertical direction, so that a designated well of the extraction reaction reagent box in the vertical direction can align with a filling unit of the pumping filling assembly.

7. The kit-instant preparation module of claim 5, wherein, It also includes a first three-way valve, a second three-way valve, a washing two-way valve, and a washing three-way valve. Three branches of the first three-way valve are connected to the reagent barrel to be filled, the washing liquid barrel, and the second three-way valve, respectively. The other two branches of the second three-way valve are connected to the filling pump and the pumping filling assembly, respectively. Two branches of the washing two-way valve are connected to the washing tank and the washing three-way valve, respectively. The other two branches of the washing three-way valve are connected to the washing pump and the washing liquid barrel, respectively.

8. The kit-instant preparation module of claim 2, wherein, The reagent bucket bearing part comprises a plurality of reagent bucket receiving positions, each of which is provided with a corresponding gravity sensing unit. The reagent bucket receiving position can bear a reagent bucket to be filled. A bottle cap unit capable of lifting motion is arranged above the reagent bucket to be filled. The bottle cap unit can be covered on the bucket opening of the reagent bucket to be filled. The bottle cap unit is provided with a reagent needle capable of communicating the inside of the reagent bucket to be filled with the liquid path of the pumping filling assembly. A bottle cap buffer is arranged at the position of the bucket opening corresponding to the reagent bucket to be filled. The bottle cap buffer can form a buffer support for the descending bottle cap unit.

9. A method of instantizing a kit, characterized by, The method comprises the following steps: The pumping step: starting the pumping filling assembly to pump a preset volume of reagent to be filled into the designated hole position of the reagent box; wherein the magnetic bead diluent filling part in the pumping filling assembly pumps a second volume of magnetic bead diluent into the designated hole position in the reagent box. The pipetting step: starting the pipetting assembly to transfer a preset volume of pipetting reagent to the designated hole position containing the reagent to be filled, and the preset volume of pipetting reagent is less than the preset volume of reagent to be filled; wherein the pipetting assembly transfers a first volume of concentrated magnetic bead mixture to the hole position containing the second volume of magnetic bead diluent; the second volume is not less than three times the first volume.

10. The kit instant preparation method according to claim 9, wherein In the pumping step, the lysis liquid filling part in the pumping filling assembly is started to pump a fourth volume of lysis liquid into the designated hole position in the reagent box; in the pipetting step, the magnetic bead mixture shaking assembly is started to shake the magnetic bead mixture reagent bottle, so that the magnetic bead mixture in the magnetic bead mixture reagent bottle produces vortex effect to mix the magnetic beads; the pipetting assembly is started to transfer a first volume of magnetic bead mixture from the mixed magnetic bead mixture reagent bottle to the hole position containing the second volume of magnetic bead diluent, and transfer a third volume of activation liquid containing protease K and internal reference from the hole position containing the fourth volume of lysis liquid; wherein the fourth volume is not less than three times the third volume.

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