Method for establishing a PCR reaction system

Abstract

The application provides a PCR reaction system establishment method, relates to the technical field of nucleic acid extraction, and comprises the following steps: displacement and sample adding of a lysis kit: in the sample adding step, in the first round, a plurality of pipettes of a variable-distance pipetting device are used to suck samples in sample tubes at least one sample tube interval from sample tube 1, and then the samples are poured into the column hole positions at least one hole interval from hole position 1, wherein the interval sample tube number is the same as the interval hole number; in the second round, the plurality of pipettes are used to suck samples in sample tubes at least one sample tube interval from sample tube 2, and then the samples are poured into the column hole positions at least one hole interval from hole position 2; and the third round and subsequent rounds are sequentially performed. Through the method, the technical problem that the serial numbers of biological samples before and after pipetting cannot be one-to-one corresponding in the prior art is solved, and the detection result is ensured to be one-to-one corresponding to the sample.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of nucleic acid extraction, in particular to a PCR reaction system establishment method. BACKGROUND

[0002] Nucleic acid (ribonucleic acid RNA, deoxyribonucleic acid DNA) is a basic unit representing the genetic characteristics of living organisms. Nucleic acid detection is an advanced biological detection at the molecular level, which has more advantages such as sensitivity, specificity, and no window period compared with traditional morphological detection, cytological detection, immunological detection, etc. In recent years, nucleic acid detection has gradually been recognized as a standard gold standard method, and is widely used in the detection and diagnosis of major infectious diseases such as hepatitis and AIDS, and new infectious diseases such as SARS, influenza, and hand-foot-mouth disease, and is widely used in biological and medical research.

[0003] In nucleic acid detection, the key steps are to extract and purify biological samples, and to mix the purified product with PCR (Polymerase Chain Reaction) reagent solution, and finally produce test products to establish a PCR reaction system. Specifically, before the experiment starts, the sample tube, purification reagent kit, suction head, reagent, and PCR tube are placed in the corresponding area of the instrument. The sample tube is generally prepared in 96 pieces, and the distribution mode is 6x16, that is, it is divided into 6 rows and 16 columns, and the order is labeled from the row to the bottom, such as 1-16, 17-32. The hole position on the purification reagent kit corresponds to the sample tube, and there are also 96 pieces, and the distribution mode is 8x12, that is, it is divided into 8 rows and 12 columns, and the order is labeled from the column to the bottom, such as 1-8, 9-16. During the experiment, in order to improve the detection efficiency, multiple pipettes are generally used for pipetting. However, due to the different distances between the sample tubes and the distances between the hole positions on the purification reagent kit, the original sample tube sequence number and the hole sequence number after pipetting cannot be one-to-one corresponding, which leads to the final detection result and the biological sample cannot be one-to-one matched. SUMMARY

[0004] The purpose of the present application is to provide a PCR reaction system establishment method to alleviate the technical problem that the sequence numbers of biological samples before and after pipetting cannot be one-to-one corresponding in the related art.

[0005] In order to solve the above technical problems, the technical means adopted by the present application is:

[0006] The PCR reaction system establishment method provided by the present application comprises the following steps:

[0007] Displacement of lysis reagent kit:

[0008] The mobile device clamps the lysis kit at the lysis position and moves it to the rotating area, and the rotating device drives the lysis kit to rotate so that each column of hole positions on the lysis kit is parallel to each row of sample tubes.

[0009] The sample adding step includes:

[0010] In the first round, the multiple pipettes of the variable-distance pipetting device suck the samples in the sample tubes at least one sample tube interval, move above the lysis kit, and then spit the samples into the column hole positions at least one hole interval.

[0011] In the second round, the multiple pipettes suck the samples in the sample tubes at least one sample tube interval, and then spit the samples into the column hole positions at least one hole interval.

[0012] In the third round and the subsequent rounds, the multiple pipettes spit the samples in the remaining sample tubes into the corresponding hole positions.

[0013] Further, in the sample adding step, the pipettes are four, and the samples are 96.

[0014] In the first round, the four pipettes suck the samples in the sample tubes at least one sample tube interval, and then spit the samples into the column hole positions at least one hole interval.

[0015] In the second round, the four pipettes suck the samples in the sample tubes at least one sample tube interval, and then spit the samples into the column hole positions at least one hole interval.

[0016] In the third round, the four pipettes suck the samples in the sample tubes at least one sample tube interval, and then spit the samples into the column hole positions at least one hole interval.

[0017] In the fourth round, the four pipettes suck the samples in the sample tubes at least one sample tube interval, and then spit the samples into the column hole positions at least one hole interval.

[0018] In the fifth round and the subsequent rounds, the four pipettes spit the samples in the remaining sample tubes into the remaining column hole positions.

[0019] Further, the variable-distance pipetting device includes a variable-distance mechanism, and the variable-distance mechanism is in driving connection with the multiple pipettes.

[0020] In the sample adding step, the variable distance mechanism drives the multiple pipettes to spread out, then the multiple pipettes select the preset sample tubes and suck the samples in intervals;

[0021] In the sample discharging step, the variable distance mechanism drives the multiple pipettes to fold up, then the multiple pipettes select the preset column holes and discharge the samples in intervals.

[0022] Further, the PCR reaction system establishing method further comprises the following steps:

[0023] Consumable preparation: place six reagent kits in the respective purification positions of the purification area, place sample tubes in the sample area, place TIP heads in the TIP head area, place reagents in the reagent area, and place PCR tubes in the PCR reaction system area;

[0024] Purification: the moving device clamps the lysed reagent kit after the sample adding step and moves it back to the lysed position, and the purification device purifies the sample in the lysed reagent kit;

[0025] Liquid preparation: after the lysed reagent kit is placed back to the lysed position, the pipettes mix the reagents in the reagent area into the PCR tubes;

[0026] Elution reagent kit shifting: the moving device clamps the elution reagent kit on the elution position and moves it to the rotating area, and the rotating device drives the elution reagent kit to rotate to each column hole on the elution reagent kit and each row of sample tubes to be parallel to each other;

[0027] Liquid mixing: the multiple pipettes suck the products in the elution reagent kit in intervals and then discharge them into the PCR tubes that have completed liquid preparation in intervals.

[0028] Further, the rotating area is between the purification area and the sample area;

[0029] From the rotating area to the direction away from the rotating area and the sample area, the respective purification positions are distributed in intervals;

[0030] The reagent kit placed on the purification position has each row of holes parallel to each row of sample tubes in the sample area.

[0031] Further, the TIP head area, the reagent area, and the PCR reaction system area are arranged on the side of the sample area away from the rotating area, and along the length direction of the sample area, the three areas are distributed in sequence.

[0032] Further, the rotating device comprises a rotating driving member and a fixed table;

[0033] The rotating driving member is in transmission connection with the fixed table;

[0034] The fixed platform is used to place the reagent kit.

[0035] Furthermore, the moving device includes a lifting mechanism and a gripping mechanism;

[0036] The lifting mechanism is connected to the clamping mechanism for driving the clamping mechanism to move longitudinally;

[0037] The gripping mechanism is used to grip the reagent kit.

[0038] Furthermore, the gripping mechanism includes an electric gripper and a gripper hand;

[0039] The gripper has two grippers, which are correspondingly arranged on the two grippers of the electric gripper, so that they can move towards or away from each other during the operation of the two grippers.

[0040] Furthermore, the mobile device also includes a first moving mechanism and a second moving mechanism;

[0041] The first moving mechanism is disposed on the frame and can travel along the length of the frame;

[0042] The second moving mechanism is disposed on the first moving mechanism and is connected to the variable-pitch pipetting device for driving the variable-pitch pipetting device to move along the width direction of the frame.

[0043] Compared with existing technologies, the beneficial effects of the PCR reaction system establishment method provided by this invention are as follows:

[0044] In this application, before sample addition, the moving and rotating devices work together to move the lysis kit to the rotating area, ensuring that each row of wells in the lysis kit is parallel to each row of sample tubes. This ensures that when samples are aspirated row by row and dispensed column by column, the number of wells added is an integer after each row of samples is aspirated, facilitating accurate matching after sample pipetting. During sample addition, in the first round, multiple pipettes aspirate samples from the corresponding row of sample tubes, starting from sample tube 1 and skipping at least one sample tube. Then, the aspirated samples are aspirated from well 1, skipping at least one well. In the first round, multiple pipettes, starting from sample tube 2 and skipping at least one sample tube, aspirate samples from the sample tubes in the same row. After aspiration, starting from well 2 and skipping at least one well, the pipettes aspirate samples from the wells in the same row. This process is repeated in the third round and thereafter. Here, the number of sample tubes skipped is the same as the number of wells skipped, and the number of skips in each round is also the same as in the previous round. This ensures that the original sample tube number corresponds one-to-one with the well number after pipetting, thus ensuring a one-to-one correspondence between the sample and the test result. Attached Figure Description

[0045] In order to make the technical solutions in the specific embodiments or related art of the present application clearer, the accompanying drawings needed in the specific embodiments or related art description will be briefly introduced as follows. Obviously, the accompanying drawings in the following description are some embodiments of the present application, and other accompanying drawings can be obtained by those skilled in the art without any creative effort on the basis of these accompanying drawings.

[0046] Figure 1 The structural schematic diagram of the nucleic acid purification instrument provided by the embodiment of the present application is shown in the figure.

[0047] Figure 2 The top view of each area of the nucleic acid purification instrument provided by the embodiment of the present application is shown in the figure.

[0048] Figure 3 The structural schematic diagram of the rotating device of the nucleic acid purification instrument provided by the embodiment of the present application is shown in the figure.

[0049] Figure 4 The application schematic diagram of the moving device of the nucleic acid purification instrument provided by the embodiment of the present application is shown in the figure.

[0050] Figure 5 The enlarged view of the I of the figure is shown in the figure. Figure 4

[0051] The enlarged view of the II of the figure is shown in the figure. Figure 6 Figure 4 The transmission schematic diagram of the second moving mechanism and the variable-distance pipetting device of the nucleic acid purification instrument provided by the embodiment of the present application under a certain viewing angle is shown in the figure.

[0052] Figure 7 The transmission schematic diagram of the second moving mechanism and the variable-distance pipetting device of the nucleic acid purification instrument provided by the embodiment of the present application under another viewing angle is shown in the figure.

[0053] Figure 8 The transmission schematic diagram of the lifting mechanism and the clamping mechanism of the nucleic acid purification instrument provided by the embodiment of the present application is shown in the figure.

[0054] Figure 9 The icon is shown in the figure.

[0055] The icon is shown in the figure.

[0056] ​100 - mobile device; 110 - lifting mechanism; 120 - clamping mechanism; 130 - first moving mechanism; 140 - second moving mechanism; 111 - fixed frame; 112 - lifting motor; 113 - screw rod; 114 - transmission nut; 115 - sliding connecting plate; 121 - electric clamping jaw; 122 - clamping hand; 131 - first rotary driver; 132 - first driving wheel; 133 - first driven wheel; 134 - first synchronous belt; 135 - X-direction sliding frame; 136 - first fixed plate; 141 - second rotary driver; 142 - second driving wheel; 143 - second driven wheel; 144 - second synchronous belt; 145 - second fixed plate;

[0057] 200 - lysis kit; 300 - rotating area;

[0058] 400 - rotating device; 410 - rotating driving part; 420 - fixed table; 430 - limiting block; 440 - fixed support;

[0059] 500 - variable distance pipetting device; 600 - purification area; 700 - sample area; 800 - TIP head area; 900 - reagent area; 1000 - PCR reaction system area; 1100 - purification device; 1200 - elution kit; 1300 - rack; 1400 - bottom plate. DETAILED DESCRIPTION

[0060] The technical solutions of the present application will be described clearly and completely below in conjunction with the drawings. Obviously, the described embodiments are part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0061] In the description of the present application, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer" and the like indicate the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present application and simplifying the description, and do not indicate or imply that the indicated device or element must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as a limitation on the present application. In addition, the terms "first", "second", "third" are only for the purpose of description, and cannot be understood as indicating or implying relative importance.

[0062] In the description of the present application, it should be noted that unless specifically defined and limited otherwise, the terms "mounting", "connecting", "connecting" should be understood broadly, for example, it can be fixedly connected, or it can be detachably connected, or integrally connected; it can be mechanically connected, or it can be electrically connected; it can be directly connected, or it can be indirectly connected through an intermediate medium, or it can be connected inside two elements. For those skilled in the art, the specific meaning of the above terms in the present application can be understood according to the specific circumstances.

[0063] The PCR reaction system establishment method provided by the embodiment comprises the following steps:

[0064] S1, consumable preparation: place six reagent kits on the purification sites of the purification area 600, place the sample tubes on the sample area 700, place the TIP head on the TIP head area 800, place the reagents on the reagent area 900, and place the PCR tubes on the PCR reaction system area 1000;

[0065] S2, reagent box displacement: the moving device 100 clamps the lysis reagent box 200 on the lysis site and moves it to the rotating area 300, and the rotating device 400 drives the lysis reagent box 200 to rotate to each column of hole sites on the lysis reagent box 200 and each row of sample tubes are parallel to each other;

[0066] S3, sample addition: in the first round, the multiple pipettes of the variable-distance pipetting device 500 start from the sample tube 1 and at least interval one sample tube to suck the sample in the row of sample tubes, and after the sample is sucked, it is moved to above the lysis reagent box 200, and at least interval one hole site to spit the sample into the column hole site from hole site 1, wherein the interval sample tube number and the interval hole site number are the same; in the second round, the multiple pipettes start from the sample tube 2 and at least interval the same number of sample tubes as the first round to suck the sample in the row of sample tubes, and after the sample is sucked, the sample is spit into the column hole site from hole site 2 with the same number of hole sites as the first round interval; in the third round and later, the multiple pipettes add the samples in the remaining sample tubes to the corresponding hole sites in the above manner;

[0067] S4, purification: the moving device 100 clamps the lysis reagent box 200 which has completed the sample addition in the sample addition step, and moves it back to the lysis site, and the purification device 1100 purifies the sample in the lysis reagent box 200;

[0068] S5, liquid preparation: after the lysis reagent box 200 is placed back to the lysis site, the pipettes mix the reagents in the reagent area 900 into the PCR tubes;

[0069] S6, elution kit displacement: the moving device 100 picks up the elution kit 1200 in the elution position and moves it to the rotating area 300, and the rotating device 400 drives the elution kit 1200 to rotate to each column of hole positions on the elution kit 1200 and each row of sample tubes are parallel to each other;

[0070] S7, mixing: multiple pipettes take the product in the elution kit 1200 at intervals, and then spit it into the PCR tube that has completed the liquid preparation at intervals.

[0071] Reference Figure 1 and Figure 2 The rotating area 300 is between the purification area 600 and the sample area 700, from the rotating area 300 towards the direction away from the rotating area 300 and the sample area 700, each purification position is distributed at intervals, and the kit placed on the purification position has each row of hole positions parallel to each row of sample tubes in the sample area 700; the T I P head area 800, the reagent area 900 and the PCR reaction system area 1000 are arranged on the side of the sample area 700 away from the rotating area 300, and along the length direction of the sample area 700, the three are arranged in sequence, wherein the T I P head area 800 is provided with two hole plates, the number of hole positions on the hole plate is the same as that of the kit, but each column of hole positions on the hole plate is parallel to each row of sample tubes, and the T I P head is arranged at the corresponding hole position on the hole plate; the reagent area 900 is provided with a hole plate, and the hole plate is placed with each reagent and part of the T I P head; the PCR reaction system area 1000 is provided with a hole plate, the number of hole positions on the hole plate is the same as that of the kit, but each column of hole positions on the hole plate is parallel to each row of sample tubes, and the PCR tube is arranged at the corresponding hole position on the hole plate.

[0072] The above arrangement makes the layout of consumables more compact, the space utilization rate is high, and when adding samples, preparing liquids and mixing, the displacement of the variable-distance pipetting device 500 is small, thereby reducing the time for obtaining the final PCR reaction system.

[0073] After the consumables are placed, the moving device 100 and the rotating device 400 cooperate to move the lysis kit 200 to the rotating area 300, and make each column of hole positions of the lysis kit 200 parallel to each row of sample tubes, so that when the samples are taken by row and spit by column, the number of hole positions for adding samples is an integer after taking each row of samples, thereby facilitating the corresponding after the sample pipetting.

[0074] When adding samples, in the first round, multiple pipettes start from sample tube 1 and suck samples in the sample tubes in the same row at least one sample tube interval, and then spit the samples into the wells in the same column at least one well interval from well 1. In the second round, multiple pipettes start from sample tube 2 and suck samples in the sample tubes in the same row at least one sample tube interval, and then spit the samples into the wells in the same column at least one well interval from well 2. In the third round and the subsequent rounds, the same method is used. Here, the number of sample tube intervals is the same as the number of well intervals, and the interval number of the subsequent round is the same as that of the previous round. In this way, the sample tube sequence number before sample transfer is one-to-one corresponding to the well sequence number after sample transfer, thereby ensuring one-to-one correspondence between the sample and the detection result.

[0075] When purifying, the purification device 1100 extracts DNA, RNA, and final nucleic acid through steps such as heating, lysis, horizontal shaking, vertical shaking, washing, and elution. The purified product is in the elution kit 1200. The liquid preparation step is performed in parallel with the purification step. After the reagents in the reagent area 900 are mixed into the PCR tube by the pipette, the PCR reagent liquid preparation is completed. After the purification is completed, the moving device 100 and the rotating device 400 cooperate to move the elution kit 1200 containing the purified product to the rotating area 300, and make each column of wells of the elution kit 1200 parallel to each row of sample tubes. Referring to Figure 2 At this time, the elution kit 1200 corresponds to the well plate in which the PCR tube is placed, that is, the columns of the two are parallel to each other. Multiple pipettes can sequentially add the purified product to the PCR tube to produce the test product and establish a PCR reaction system, and the test product in each PCR tube can find the corresponding sample.

[0076] It should be noted here that in the sample adding step and the liquid mixing step, the pipette needs to be equipped with a TIP head first, that is, the pipette needs to be moved to the TIP head area 800 first, and then the sample or the purified product is sucked after the TIP head is vertically inserted and tightened. After spitting into the well, the TIP head is removed and a new TIP head is reassembled for the next suction. In the liquid preparation step, the pipette needs to be moved to the reagent area 900 first, and then the TIP head of the reagent area 900 is assembled. Then the reagent is sucked and spit into the PCR tube. Similarly, a new TIP head needs to be assembled for the next suction.

[0077] In an embodiment of the present application, referring to Figure 2 and Figure 7 The sample has 96 samples, which are arranged in sample tubes arranged in 6x16, the kit is a 96-well plate, the number of wells is arranged in 8x12, and the pipette has four pipettes.

[0078] When adding samples, in the first round, four pipettes start to suck samples from sample tubes 1, interval one sample tube, and then spit the samples into the corresponding column hole, interval one hole; in the second round, four pipettes start to suck samples from sample tubes 2, interval one sample tube, and then spit the samples into the corresponding column hole, interval one hole; in the third round, four pipettes start to suck samples from sample tubes 9, interval one sample tube, and then spit the samples into the corresponding column hole, interval one hole; in the fourth round, four pipettes start to suck samples from sample tubes 10, interval one sample tube, and then spit the samples into the corresponding column hole, interval one hole; in the fifth round and later, four pipettes add the remaining sample tubes to the remaining column hole in the above-mentioned manner. It can be seen that the above-mentioned sample adding process ensures that the sample serial number and the 96-hole plate hole serial number are one-to-one corresponding, and ensures that the sample and the final result are one-to-one matched. In addition, the sample adding process is also applicable to other specifications of hole plates and sample numbers.

[0079] Reference Figure 7 The variable-distance pipetting device 500 comprises a variable-distance mechanism, which is in driving connection with the plurality of pipettes; in the sample adding step, when sucking the sample, the variable-distance mechanism drives the plurality of pipettes to expand, and then the plurality of pipettes select a preset row of sample tubes and suck the sample intervally; when spitting the sample, the variable-distance mechanism drives the plurality of pipettes to fold, and then the plurality of pipettes select a preset column of hole positions and spit the sample intervally.

[0080] By using the variable-distance pipetting device 500, when the plurality of pipettes expand, the pipettes can suck the sample interval one sample tube, and when the plurality of pipettes fold, the pipettes can spit the sample interval one hole, so as to ensure that the sample serial number, the hole serial number and the PCR tube serial number are one-to-one corresponding in the establishment of the PCR reaction system.

[0081] Further, reference Figure 3 The rotating device 400 comprises a rotating driving member 410 and a fixed table 420; the rotating driving member 410 is in driving connection with the fixed table 420; the fixed table 420 is used for placing the reagent box.

[0082] Specifically, continuing to refer to Figure 3 The rotating device 400 further comprises a fixed support 440, which is installed on the bottom plate 1400; the rotating driving member 410 adopts a motor, which is fixedly installed on the fixed support 440; the fixed table 420 is fixed to the output shaft of the motor to rotate synchronously with the output shaft; the fixed table 420 is provided with a plurality of limiting blocks 430, which are used for limiting the translational degree of freedom of the reagent box in the horizontal direction.

[0083] Reference Figure 2, through the rotating device 400, in the displacement step, the lysis kit 200 can be rotated 90°, so that each column of hole positions thereof is parallel to each row of sample tubes, so as to ensure the correspondence of the sample and the hole position after sample addition; the elution kit 1200 can be rotated 90°, so as to correspond to the hole plate on which the PCR tube is placed, so as to ensure the correspondence of the sample and the PCR tube after mixing. In addition, after the sample addition is completed, the rotating device 400 needs to drive the lysis kit 200 to rotate 90° again, so as to facilitate the clamping of the moving device 100.

[0084] Further, referring to Figures 4 to 9 , the moving device 100 comprises a first moving mechanism 130, a second moving mechanism 140, a lifting mechanism 110 and a clamping mechanism 120; wherein the first moving mechanism 130 is arranged on the rack 1300 and can travel on the rack 1300 along the length direction of the rack 1300; the second moving mechanism 140 is arranged on the first moving mechanism 130 and is in driving connection with the variable-distance pipetting device 500, for driving the variable-distance pipetting device 500 to move along the width direction of the rack 1300; the lifting mechanism 110 is arranged on the first moving mechanism 130 to travel with the first moving mechanism 130; the lifting mechanism 110 is also in driving connection with the clamping mechanism 120, for driving the clamping mechanism 120 to move along the longitudinal direction, and the clamping mechanism 120 is used for clamping the reagent kit.

[0085] Specifically, referring to Figures 4 to 6 , the first moving mechanism 130 comprises a first rotating driver 131, a first driving wheel 132, a first driven wheel 133, a first synchronous belt 134, an X-direction sliding frame 135 and a first fixed plate 136; wherein the first rotating driver 131 is an electric motor and is fixed to the rack 1300, the first driving wheel 132 is fixed to the output end of the electric motor, the first driven wheel 133 is in rotational cooperation with the rack 1300, the first synchronous belt 134 is tensioned to the first driving wheel 132 and the first driven wheel 133, the X-direction sliding frame 135 is in sliding connection with the rack 1300, and the first fixed plate 136 has two, the two first fixed plates 136 are clamped and fixed to the first synchronous belt 134, and are also fixedly connected with the X-direction sliding frame 135. When the electric motor is started, the first synchronous belt 134 rotates and synchronously drives the X-direction sliding frame 135 to move, that is, the X-direction sliding frame 135 realizes the movement along the X-direction of the rack 1300.

[0086] Referring to Figure 7 and Figure 8, the second moving mechanism 140 comprises a second rotating driver 141, a second driving wheel 142, a second driven wheel 143, a second synchronous belt 144 and two second fixing plates 145; wherein the second rotating driver 141 is an electric motor fixed to the X-direction sliding frame 135, the second driving wheel 142 is fixed to the output end of the electric motor, the second driven wheel 143 is pivoted to the X-direction sliding frame 135, the second synchronous belt 144 is tensioned between the second driving wheel 142 and the second driven wheel 143, and the two second fixing plates 145 are clamped and fixed to the second synchronous belt 144 and are further fixedly connected to the variable-distance pipetting device 500. When the electric motor is started, the second synchronous belt 144 rotates and synchronously drives the variable-distance pipetting device 500 to move, i.e. the variable-distance pipetting device 500 realizes movement along the Y-direction of the rack 1300, and at the same time, it can also move along the X-direction of the sliding frame 135. Thus, under the cooperation of the first moving mechanism 130 and the second moving mechanism 140, the variable-distance pipetting device 500 can move in the horizontal plane to realize sample adding, liquid preparation and liquid mixing.

[0087] Reference Figure 9 , the lifting mechanism 110 comprises a fixed frame 111, a lifting motor 112, a lead screw 113, a transmission nut 114 and a sliding connection plate 115; wherein the fixed frame 111 is mounted to the X-direction sliding frame 135, so that the lifting mechanism 110 can move along the X-direction of the rack 1300, the lifting motor 112 is fixed to the top end of the fixed frame 111, the lead screw 113 is connected with the output shaft of the lifting motor 112, the transmission nut 114 is threadedly connected with the lead screw 113, and the sliding connection plate 115 is slidingly connected with the fixed frame 111 and is fixedly connected with the transmission nut 114. Thus, when the lifting motor 112 is started, the lead screw 113 rotates, and the sliding connection plate 115 slides up and down with the transmission nut 114.

[0088] Reference Figure 9 , the clamping mechanism 120 comprises an electric clamping jaw 121 and two clamping hands 122; the electric clamping jaw 121 is fixed to the sliding connection plate 115, and the two clamping hands 122 are correspondingly arranged at the two clamping jaws of the electric clamping jaw 121 to move towards or away from each other. When the electric clamping jaw 121 is started, the clamping hands 122 can clamp and release the reagent box. In addition, through the first moving mechanism 130 and the lifting mechanism 110, the clamping jaw can move along the X-direction and Z-direction of the rack 1300 to realize clamping and placing of the reagent box.

[0089] It should be pointed out finally that the above embodiments are only used to illustrate the technical solutions of the present application, and are not intended to limit the present application. Although the present application has been described in detail with reference to the above embodiments, those skilled in the art should understand that the technical solutions recorded in the above embodiments can be modified, or some or all of the technical features can be replaced by equivalents, and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present application.

Claims

1. A method for establishing a PCR reaction system, characterized by, The method comprises the following steps: The lysis kit is moved: The moving device clamps the lysis kit on the lysis site and moves it to the rotating area, and the rotating device drives the lysis kit to rotate to the state that each column of hole sites on the lysis kit is parallel to each row of sample tubes; The sample is added: The pipettes of the variable-distance pipetting device have four pipettes, and the sample has 96 samples, which are arranged in 16 rows and 6 columns. After the lysis kit is rotated, it is arranged in 8 rows and 12 columns. In the first round, the four pipettes of the variable-distance pipetting device start from the first column and the first sample tube, and suck the sample in the sample tube in the column at intervals of one sample tube. After the sample is sucked, the pipettes are moved above the lysis kit, and the sample is poured into the hole site in the column from the first hole site in the first column at intervals of one hole site. The number of interval sample tubes is the same as the number of interval hole sites. In the second round, the four pipettes start from the second sample tube in the column and suck the sample in the sample tube in the column at intervals of one sample tube. After the sample is sucked, the pipettes start from the second hole site in the first column and pour the sample into the hole site in the column at intervals of one hole site. In the third round, the four pipettes start from the ninth sample tube in the column and suck the sample in the sample tube in the column at intervals of one sample tube. After the sample is sucked, the pipettes start from the first hole site in the second column and pour the sample into the hole site in the column at intervals of one hole site. In the fourth round, the four pipettes start from the tenth sample tube in the column and suck the sample in the sample tube in the column at intervals of one sample tube. After the sample is sucked, the pipettes start from the second hole site in the second column and pour the sample into the hole site in the column at intervals of one hole site. In the fifth round and later, the four pipettes pour the remaining samples in the sample tubes into the hole sites in the remaining columns of the lysis kit in the order of the first round to the fourth round. The method for establishing the PCR reaction system further comprises the following steps: Preparation of consumables: six kits are placed in the purification sites of the purification area, sample tubes are placed in the sample area, TIP heads are placed in the TIP head area, reagents are placed in the reagent area, and PCR tubes are placed in the PCR reaction system area. Purification: the moving device clamps the lysis kit after the sample adding step is completed, and moves it back to the lysis site. The purification device purifies the sample in the lysis kit. Liquid preparation: after the lysis kit is placed back to the lysis site, the pipettes mix the reagents in the reagent area into the PCR tubes. The elution kit is moved: Mixing: multiple pipettes suck the product in the elution kit at intervals, and then pour it into the PCR tubes that have completed liquid preparation at intervals. The rotating area is between the purification area and the sample area; From the rotating area to the direction away from the rotating area and the sample area, each purification site is distributed at intervals; The hole sites in each row of the kit placed in the purification site are parallel to each row of sample tubes in the sample area.

2. The PCR reaction system establishment method according to claim 1, characterized in that, The variable-distance pipetting device comprises a variable-distance mechanism, which is in transmission connection with the plurality of pipetting guns; In the sample adding step, when the samples are sucked, the variable-distance mechanism drives the plurality of pipetting guns to spread out, and then the plurality of pipetting guns select the preset sample tubes in the row and suck the samples at intervals; In the sample discharging step, when the samples are discharged, the variable-distance mechanism drives the plurality of pipetting guns to fold, and then the plurality of pipetting guns select the preset sample wells in the column and discharge the samples at intervals.

3. The PCR reaction system establishment method according to claim 1, characterized in that, The TIP head area, the reagent area and the PCR reaction system area are arranged on the side of the sample area away from the rotating area, and along the length direction of the sample area, the three areas are sequentially distributed.

4. The PCR reaction system establishment method according to claim 1, characterized in that, The rotating device comprises a rotating driving member and a fixed table; The rotating driving member is in transmission connection with the fixed table; The fixed table is used for placing the reagent box.

5. The PCR reaction system establishment method according to claim 1, characterized in that, The moving device comprises a lifting mechanism and a clamping mechanism; The lifting mechanism is in transmission connection with the clamping mechanism, and is used for driving the clamping mechanism to move along the longitudinal direction; The clamping mechanism is used for clamping the reagent box.

6. The PCR reaction system establishment method according to claim 5, characterized in that, The clamping mechanism comprises an electric clamping jaw and a clamping hand; The clamping hand has two, and is correspondingly arranged on the two clamping jaws of the electric clamping jaw, so as to move towards or away from each other in the action of the two clamping jaws.

7. The PCR reaction system establishment method according to claim 5, characterized in that, The moving device further comprises a first moving mechanism and a second moving mechanism; The first moving mechanism is arranged on the rack and can travel on the rack along the length direction of the rack; The second moving mechanism is arranged on the first moving mechanism and is in transmission connection with the variable-distance pipetting device, and is used for driving the variable-distance pipetting device to move along the width direction of the rack.

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