Specific protein analyzer for tumor immunity assessment and tumor immunity assessment method

Abstract

The present invention provides a specific protein analyzer for tumor immunity assessment and a tumor immunity assessment method, including a support seat and a reagent tube, the top surface of the support seat is provided with a transfer box, the rear wall of the transfer box is provided with a detection mechanism, the detection mechanism and the inside of the transfer box are connected through, a transfer assembly is provided inside the transfer box, the top surface of the support seat is a concave structure, the concave structure of the support seat is provided with a sample placement assembly, a plurality of reagent tubes are inserted and placed on the top surface of the sample placement assembly, the top of the reagent tube is closed and connected with a sealing plug, and a separation mechanism is provided on one side of the support seat. The present invention transfers the reagent tubes one by one through the transfer assembly, uses the detection mechanism to extract the human plasma in the test reagent tube, and the remaining reagent tube and the human plasma inside continue to be transferred between the lower through assembly and the upper through assembly, and the lower through assembly is used to penetrate the bottom of the reagent tube, so that the remaining human plasma flows out to the waste liquid bag, thereby improving the convenience of separating the reagent tube and the human plasma.

Description

Technical Field

[0001] The present invention relates to the technical field of biological detection equipment, and in particular to a specific protein analyzer for evaluating tumor immunity and a tumor immunity evaluation method. Background Art

[0002] Traditional methods of early cancer detection include: imaging, serology, and genetic testing. However, imaging is mostly used for clinical diagnosis and cannot detect cancer factors early. The detection period is in the middle and late stages. Serology is inaccurate, with an accuracy of about 50% to 60%, and the rates of missed detection, false positives, and false negatives are extremely high. Genetic testing is difficult to promote, and has the characteristics of complex operating techniques and high prices. From this point of view, compared with traditional treatment methods, tumor immunity assessment is the most effective means of preventing and treating cancer. It allows people to understand their immunity level and immune status at any time, master their physical health status, and enable people to adjust their lifestyles in time to prevent diseases before they occur. It can also provide important quantitative basis for patients with various tumors, such as disease diagnosis, medication guidance, and prognosis.

[0003] In tumor immunity assessment, it is necessary to use a reagent tube with a sealed stopper to collect human peripheral blood, use a specific protein analyzer to puncture the sealed stopper to extract the plasma in the reagent tube, and detect the complement, immunoglobulin IgE, immunoglobulin IgA, immunoglobulin IgG and immunoglobulin IgM expression in the plasma. According to the deviation between the test value and the normal detection range, the relevant results are given, and then the tested reagent tube is transported for recycling. However, this specific protein analyzer has the following problems in recycling:

[0004] After testing human plasma, the reagent tube with human plasma will continue to be transferred without separating the human plasma in the reagent tube, resulting in the inability to directly recycle the reagent tube. The sealing plug of the reagent tube needs to be opened manually to pour out the human plasma. This separation and recovery method is very cumbersome and the incomplete separation of the reagent tube and the residual liquid directly leads to: ① Sensitivity limitation in subsequent tumor immunity evaluation: Although the specific protein analyzer can detect a variety of specific proteins, its sensitivity is not enough to detect trace protein changes in the tumor microenvironment. This is because the reagent tube and the residual liquid are not completely separated, especially in the early stage of tumor or minimal residual disease (MRD). ② The standardization of scoring in subsequent tumor immunity evaluation: Specific protein analyzers from different manufacturers may use different detection methods and standards, and because of the convenience of separating the reagent tube and human plasma in manual operation, the comparability of the subsequent scoring results is affected. In addition, the operating standards and quality control measures between different laboratories may be inconsistent, which will also affect the accuracy and reliability of the results. Therefore, it is necessary to control the convenience of separating the reagent tube and human plasma in manual operation to ensure the complete separation of the reagent tube and the residual liquid.

[0005] In summary, there is a need for a specific protein analyzer that is easy to operate for metastasis detection and processing, as well as a systematic method for evaluating tumor immunity. Summary of the invention

[0006] In view of the deficiencies of the prior art, the present invention provides a specific protein analyzer for tumor immunity assessment and a tumor immunity assessment method, which solves the problems mentioned in the background technology.

[0007] To achieve the above objectives, the present invention is implemented through the following technical solutions:

[0008] A specific protein analyzer for evaluating tumor immunity comprises a support base and a reagent tube, wherein a transfer box is arranged on the top surface of the support base, a detection mechanism is arranged on the rear wall of the transfer box, the detection mechanism and the interior of the transfer box are connected through, a transfer assembly is arranged inside the transfer box, the top surface of the support base is a concave structure, a sample placement assembly is arranged inside the concave structure of the support base, a plurality of reagent tubes are inserted and placed on the top surface of the sample placement assembly, a sealing plug is connected to the top of the reagent tube, a separation mechanism is arranged on one side of the support base, a first groove is provided inside the support base, a second groove is provided on the bottom surface of the support base, the separation mechanism is arranged on the top of the first groove and the second groove, a waste liquid bag is inserted inside the second groove, a recovery bag is inserted on the front wall of the support base, a pushing assembly and a second pneumatic rod are arranged on the rear wall of the transfer box, the sample placement assembly is transferred to the bottom of the separation mechanism through the cooperation of the pushing assembly and the second pneumatic rod, a control chassis is fixed on the front wall of the transfer box, and a touch screen is arranged on the front wall of the control chassis;

[0009] The detection mechanism includes a detection box, an extraction component, an analysis device, a cleaning device and an air drying device. The detection box is connected to the rear wall of the transfer box. An extraction component is arranged inside the detection box. The extraction component is used to extract the reagent tube transferred by the transfer component. The extraction component is sequentially arranged with an analysis device, a cleaning device and an air drying device in the circumferential direction.

[0010] The separation mechanism includes a fourth motor, a gear, a lower-through component and an upper-through component. The lower-through component is inserted into the first groove, and the upper-through component is inserted into the top surface of the support seat. The fourth motor is arranged on the top surface of the support seat. The rotating end of the fourth motor is connected to a gear, and both sides of the gear are respectively meshed and connected between the upper-through component and the lower-through component. The lower-through component is used to puncture the bottom end of the reagent tube to release the residual liquid, and the upper-through component is used to stop the reagent tube from rising.

[0011] As a second aspect of the invention, a method for evaluating tumor immunity is proposed, comprising the following steps:

[0012] S1. Data Collection

[0013] S1-1, obtaining sample data of the questionnaire, wherein the sample data includes: lifestyle, personal medical history, family history of cancer, environmental factors and pathological data of the target patient collected by the questionnaire;

[0014] S1-2, draw 2 ml of peripheral blood from the target patient, detect the white blood cell count, neutrophil percentage and count, lymphocyte percentage and count, monocyte percentage and count, and determine the basic status of the target patient's blood cells;

[0015] S1-3, based on the specific protein analyzer, detecting the humoral immune protein in the peripheral blood of the target patient to determine the humoral immune protein;

[0016] S1-4, use flow cytometry to detect peripheral blood lymphocytes, separate and count lymphocytes from the peripheral blood of the target patient, and obtain the proportion and number of immune cells;

[0017] S2. Data preprocessing

[0018] S2-1, score the collection results obtained in each stage of S1-1, S1-2 and S1-3, and substitute the test data obtained in step S1-4 into Flowjo V10 analysis software, Excel and Grphpad prism 8.0 for data analysis and processing, and use t-test to score the data of the deviation between the test value and the normal detection range in the test data;

[0019] S2-2, use flow cytometry to detect the killing activity of NK cells in peripheral blood PBMC against tumor cells, and score according to the deviation between the detection value and the normal detection range;

[0020] S3. Calculate the immunity score:

[0021] According to the formula Perform weighted calculation on the scores obtained at each stage and output the immunity score;

[0022] Where n represents the number of items tested in the immune assessment, Pi represents the item weight, and Si represents the test value at each stage of scoring.

[0023] The present invention provides a specific protein analyzer for tumor immunity assessment and a tumor immunity assessment method. Compared with the prior art, it has the following beneficial effects:

[0024] 1. When testing the specific protein in human plasma loaded in the reagent tube, first place the reagent tube on the sample placement component, then transfer the reagent tubes one by one through the transfer component, use the detection mechanism to extract the human plasma in the test reagent tube, and the remaining reagent tube and the human plasma inside are further transferred between the lower through component and the upper through component, and the lower through component is used to penetrate the bottom of the reagent tube so that the remaining human plasma flows out to the waste liquid bag, and then the reagent tube is placed in the recovery bag, which improves the convenience of separating the reagent tube and the human plasma;

[0025] 2. The second lifting plate is lowered by driving the second tooth plate to move downward, so that the multiple third needle tubes pierce the sealing plugs and enter the reagent tubes at the same time. Then, the cleaning liquid in the water tank is introduced into the reagent tubes for cleaning by using a water pump. The cleaning solution is continuously discharged through the second needle tubes, thereby ensuring the complete separation of the reagent tubes and the residual liquid.

[0026] 3. Accurately quantifying the tumor immunity assessment based on the collection results at each stage is of great significance for the public's health management and the diagnosis and treatment of tumor populations. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0028] Figure 1 The schematic diagram of the structure of the specific protein analyzer for evaluating tumor immunity of the present invention is shown;

[0029] Figure 2 The overall back structure schematic diagram of the present invention is shown;

[0030] Figure 3 A schematic diagram of the connection structure of the top surface of the support seat of the present invention is shown;

[0031] Figure 4 The schematic diagram of the transfer box structure of the present invention is shown;

[0032] Figure 5 A schematic diagram of the structure of the sample placement component of the present invention is shown;

[0033] Figure 6 A schematic diagram of the transfer assembly structure of the present invention is shown;

[0034] Figure 7 A schematic diagram of the structure of the extraction component of the present invention is shown;

[0035] Figure 8A schematic diagram of the separation mechanism structure of the present invention is shown;

[0036] Fig. 9 It shows a schematic diagram of the structure of the lower piercing assembly and the upper piercing assembly of the present invention cooperating with the puncture reagent tube;

[0037] Fig.10 A schematic diagram of the structure of the push component of the present invention is shown;

[0038] Fig.11 A schematic diagram of the structure of the waste liquid bag of the present invention is shown;

[0039] Fig.12 A schematic diagram of the structure of the recycling bag of the present invention is shown;

[0040] Fig.13 A schematic diagram of lymphocyte subpopulations obtained by flow cytometry based on the tumor immunity assessment method proposed in the present invention is shown;

[0041] Fig.14 A schematic diagram of detecting the killing activity of NK cells against tumor cells in peripheral blood PBMC using flow cytometry is shown;

[0042] Fig.15 The figure shows a schematic flow chart of the tumor immunity evaluation method proposed in the present invention.

[0043] As shown in the figure: 1. support seat; 11. first groove; 12. second groove; 13. first pneumatic rod; 14. first push plate; 2. sample placement assembly; 21. tray; 211. third groove; 212. first limit plate; 213. puncture groove; 22. positioning plate; 3. transfer box; 31. push assembly; 311. first motor; 312. first fixing plate; 313. second push plate; 314. screw; 32. second pneumatic rod; 33. sliding interface; 34. touch screen; 35. control chassis; 36. oblique stopper; 4. detection mechanism; 41. detection box; 42. extraction assembly; 421. third pneumatic rod; 422. third push plate; 423. second motor; 424. first turntable; 425. air pump; 426. guide cylinder; 427. first needle tube; 4 3. Analytical device; 44. Cleaning device; 45. Air drying device; 5. Waste liquid bag; 6. Recovery bag; 7. Transfer assembly; 71. Second fixed plate; 72. Third motor; 74. Rotating shaft; 741. Second limit plate; 75. Sleeve; 76. Second turntable; 77. Clamping claw; 78. Fourth pneumatic rod; 8. Separation mechanism; 81. Fourth motor; 82. Gear; 83. Down-through assembly; 831. First tooth plate; 832. First lifting plate; 833. First liquid guide tube; 834. Second needle tube; 84. Up-through assembly; 841. Second tooth plate; 842. Third fixed plate; 843. Water tank; 844. Water pump; 845. Second lifting plate; 846. Second liquid guide tube; 847. Third liquid guide tube; 848. Third needle tube; 9. Reagent tube; 91. Sealing plug. DETAILED DESCRIPTION

[0044] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are clearly and completely described. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0045] Embodiment 1

[0046] In order to solve the technical problems in the background technology, the following specific protein analyzer for tumor immunity assessment and tumor immunity assessment method are provided:

[0047] As a first aspect of the present invention, Figure 1 - Fig.12As shown, the specific protein analyzer for tumor immunity assessment and the tumor immunity assessment method provided by the present invention include a support base 1 and a reagent tube 9, the top surface of the support base 1 is provided with a transfer box 3, the rear wall of the transfer box 3 is provided with a detection mechanism 4, the detection mechanism 4 and the inside of the transfer box 3 are through-connected, a transfer component 7 is provided inside the transfer box 3, the top surface of the support base 1 is a concave structure, a sample placement component 2 is provided inside the concave structure of the support base 1, a plurality of reagent tubes 9 are inserted on the top surface of the sample placement component 2, the top of the reagent tube 9 is sealed and connected with a sealing plug 91, and the support base 1 is provided with a plurality of reagent tubes 9 on the top surface. A separation mechanism 8 is arranged on the side, a first groove 11 is opened inside the support seat 1, a second groove 12 is opened on the bottom surface of the support seat 1, the separation mechanism 8 is arranged at the top of the first groove 11 and the second groove 12, a waste liquid bag 5 is inserted in the second groove 12, a recovery bag 6 is inserted in the front wall of the support seat 1, a pushing assembly 31 and a second pneumatic rod 32 are arranged on the rear wall of the transfer box 3, the sample placement assembly 2 is transferred to the bottom of the separation mechanism 8 through the pushing assembly 31 and the second pneumatic rod 32, a control chassis 35 is fixed on the front wall of the transfer box 3, and a touch screen 34 is arranged on the front wall of the control chassis 35;

[0048] The detection mechanism 4 includes a detection box 41, an extraction component 42, an analysis device 43, a cleaning device 44 and an air drying device 45. The detection box 41 is connected to the rear wall of the transfer box 3. The detection box 41 is provided with an extraction component 42 inside. The extraction component 42 is used to extract the reagent tube 9 transferred by the transfer component 7. The extraction component 42 is provided with an analysis device 43, a cleaning device 44 and an air drying device 45 in sequence in the circumferential direction.

[0049] The separation mechanism 8 includes a fourth motor 81, a gear 82, a lower-through component 83 and an upper-through component 84. The lower-through component 83 is inserted into the first groove 11, and the upper-through component 84 is inserted into the top surface of the support seat 1. The fourth motor 81 is arranged on the top surface of the support seat 1. The rotating end of the fourth motor 81 is connected to the gear 82, and the two sides of the gear 82 are respectively meshed and connected between the upper-through component 84 and the lower-through component 83. The lower-through component 83 is used to puncture the bottom end of the reagent tube 9 to release the residual liquid, and the upper-through component 84 is used to stop the reagent tube 9 from rising.

[0050] It should be noted that the following effects can be achieved according to the above structure:

[0051] Because the specific protein analyzer will continue to transfer the reagent tube with human plasma when detecting human plasma during the recovery process, without separating the human plasma in the reagent tube, the reagent tube cannot be directly recycled, which directly leads to: ① Sensitivity limitation in subsequent tumor immunity evaluation: Although the specific protein analyzer can detect a variety of specific proteins, its sensitivity is not enough to detect trace protein changes in the tumor microenvironment. This is because of the incomplete separation of the reagent tube and the residual liquid, especially in the early stage of the tumor or minimal residual disease (MRD). ② The standardization of scoring in subsequent tumor immunity evaluation: Specific protein analyzers from different manufacturers may use different detection methods and standards, and because of the convenience of separating the reagent tube and human plasma in manual operation, the comparability of the subsequent scoring results is affected. Therefore, the present invention proposes that when detecting a specific protein in human plasma loaded in a reagent tube 9, the reagent tube 9 is first placed on the sample placement component 2, and then the reagent tubes 9 are transferred one by one through the transfer component 7, and the human plasma in the test reagent tube 9 is extracted using the detection mechanism 4, while the remaining reagent tube 9 and the human plasma inside are continued to be transferred between the lower penetration component 83 and the upper penetration component 84, and the bottom of the reagent tube 9 is penetrated by the lower penetration component 83, so that the remaining human plasma flows out to the waste liquid bag 5, and then the reagent tube 9 is placed in the recovery bag 6, thereby improving the convenience of separating the reagent tube 9 and the human plasma.

[0052] In this embodiment, the down-through component 83 includes a first tooth plate 831, a first lifting plate 832, a first liquid guiding tube 833 and a second needle tube 834. The first tooth plate 831 is plugged into the support seat 1. One side of the first tooth plate 831 is meshed with the gear 82. The first lifting plate 832 is fixed to the side wall of the first tooth plate 831. The first lifting plate 832 is arranged in the first groove 11. A plurality of first liquid guiding tubes 833 are plugged into the first lifting plate 832. The top of the first liquid guiding tube 833 is connected to the second needle tube 834. The bottom of the first liquid guiding tube 833 is arranged in the waste liquid bag 5.

[0053] Based on the above technical concept, by driving the first tooth plate 831 to make the first lifting plate 832 rise, multiple second needle tubes 834 can simultaneously pierce into the reagent tube 9 to release the human plasma remaining in the reagent tube 9, which is easy to operate.

[0054] In this embodiment, the upper wear component 84 includes a second tooth plate 841, a third fixed plate 842, a water tank 843, a water pump 844, a second lifting plate 845, a second liquid guide tube 846, a third liquid guide tube 847 and a third needle tube 848. The second tooth plate 841 is plugged into the support seat 1, and one side of the second tooth plate 841 is meshed and connected between the gears 82. The third fixed plate 842 is fixed to one side of the first tooth plate 831. The top surface of the third fixed plate 842 is provided with a water tank 843, and the top surface of the water tank 843 is provided with a water pump 844. The side wall of the water tank 843 is connected to the second liquid guide tube 846, and the bottom surface of the second liquid guide tube 846 is connected with a plurality of third liquid guide tubes 847. The second lifting plate 845 is fixed to the side wall of the third fixed plate 842. The third liquid guide tube 847 is plugged into the second lifting plate 845, and the bottom end of the third liquid guide tube 847 is connected to the third needle tube 848.

[0055] By driving the second tooth plate 841 to descend, the second lifting plate 845 is lowered, so that multiple third needle tubes 848 pierce the sealing plug 91 and enter the interior of the reagent tube 9 at the same time, and then the water pump 844 is used to introduce the cleaning liquid in the water tank 843 into the reagent tube 9 for cleaning. The cleaned solution continues to be discharged through the second needle tube 834, ensuring the complete separation of the reagent tube 9 and the residual liquid.

[0056] Embodiment 2

[0057] like Figure 3 , Figure 5 , Figure 6 As shown, based on the above embodiment, this embodiment further provides the following contents:

[0058] In order to achieve the above effect, the following structure is adopted;

[0059] The sample placement assembly 2 includes a tray 21 and a positioning plate 22. A plurality of third grooves 211 are provided on the top surface of the tray 21. A first limiting plate 212 is fixed to the inner wall of the third groove 211. The positioning plate 22 is inserted into the third groove 211. The bottom surface of the positioning plate 22 is attached to the top surface of the first limiting plate 212. A through hole is provided inside the positioning plate 22. The through hole of the positioning plate 22 is used for inserting the reagent tube 9. A puncture groove 213 is provided through the bottom surface of the third groove 211.

[0060] By placing the reagent tube 9 on the positioning plate 22, when extracting the plasma in the reagent tube 9, the reagent tube 9 can be transferred out by simply pushing the positioning plate 22 out of the tray 21, and after the reagent tube 9 is transferred out, the positioning plate 22 is continuously transferred, so that the positioning plate 22 continues to receive the extracted reagent tube 9, and the positioning plate 22 is inserted into another tray 21, thereby improving the convenience of transferring the reagent tube 9 and improving the convenience of transfer;

[0061] A puncture groove 213 is directly opened at the bottom of the tray 21, so that when the reagent tube 9 is transferred to the top of the lower penetration component 83, the second needle tube 834 can directly pass through the puncture groove 213 to puncture the reagent tube 9, which not only ensures the transfer of the reagent tube 9, but also ensures the separation of the reagent tube 9 and the residual liquid.

[0062] In the present embodiment, the transfer assembly 7 comprises a second fixed plate 71, a third motor 72, a rotating shaft 74, a sleeve 75, a second rotary disk 76, a clamping claw 77 and a fourth pneumatic rod 78. The second fixed plate 71 is fixed to the inner wall of the transfer box 3. The third motor 72 is arranged on the top surface of the second fixed plate 71. The rotating top end of the third motor 72 is connected to the rotating shaft 74. A second limiting plate 741 is fixed to the side wall of the rotating shaft 74. A sleeve 75 is sleeved on the top end of the rotating shaft 74. A second rotary disk 76 is fixed to the top end of the sleeve 75. The top surface of the second rotary disk 76 is connected to the fourth pneumatic rod 78. The fourth pneumatic rod 78 is connected to the inner top surface of the transfer box 3. A clamping claw 77 is fixed on the top surface of the second rotary disk 76. Four clamping claws 77 are arranged in a cross distribution.

[0063] The second turntable 76 is driven to rotate by the third motor 72, and the fourth pneumatic rod 78 is used to drive the second turntable 76 to rise and fall, so as to control the clamping claw 77 to clamp the reagent tube 9, separate the reagent tube 9 from the positioning plate 22 upward, and then rotate the reagent tube 9 to the extraction component 42 position for extraction and detection, and then continue to rotate to transfer the reagent tube 9, and the transfer operation is simple.

[0064] In this embodiment, the transfer assembly 7 further includes an electric roller 73, which is arranged on the bottom surface of the second fixed plate 71, and four electric rollers 73 are arranged in a rectangular distribution. A first pneumatic rod 13 is arranged on the top surface of the support seat 1, and a first push plate 14 is connected to the telescopic end of the first pneumatic rod 13. The electric roller 73 and the first push plate 14 are used to push the positioning plate 22 to translate;

[0065] When transferring the reagent tube 9 to the position of the claw 77, the positioning plate 22 is first pushed by the first push plate 14 so that the positioning plate 22 enters between the electric rollers 73, and then the electric rollers 73 are continued to be used to transfer the positioning plate 22, so that the claw 77 continuously transfers the reagent tube 9, and the positioning plate 22 is transferred to receive the extracted reagent tube 9, thereby improving the transfer efficiency of the positioning plate 22.

[0066] Embodiment 3

[0067] like Figure 1 , Figure 4 , Figure 7 , Fig.10 As shown, based on the above embodiment, this embodiment further provides the following contents:

[0068] In order to achieve the above effect, the following structure is adopted;

[0069] The extraction assembly 42 includes a third pneumatic rod 421, a third push plate 422, a second motor 423, a first turntable 424, an air pump 425, a guide cylinder 426 and a first needle tube 427. The third pneumatic rod 421 is arranged inside the detection box 41. The third pneumatic rod 421 is connected to the third push plate 422 at its telescopic top. The second motor 423 is arranged on the top surface of the third push plate 422. The first turntable 424 is connected to the rotating top of the second motor 423. The top surface of the first turntable 424 is provided with an air pump 425. The side wall of the air pump 425 is connected to the guide cylinder 426. Four guide cylinders 426 are arranged in a cross distribution. The bottom surface of the guide cylinder 426 is connected to the first needle tube 427.

[0070] Human plasma in the reagent tube 9 is extracted by lifting the first needle tube 427, and then passes through the analysis device 43, the cleaning device 44 and the air drying device 45 in sequence with the cooperation of the rotation of the second motor 423 and the lifting of the third pneumatic rod 421, thereby improving the convenience of detection and ensuring the accuracy of the detection results.

[0071] In this embodiment, the pushing assembly 31 includes a first motor 311, a first fixed plate 312, a second push plate 313 and a screw 314. The first fixed plate 312 is fixed to the rear wall of the transfer box 3. The first fixed plate 312 is provided with a first motor 311 on the side wall. The rotating end of the first motor 311 is connected to the screw 314. The end of the screw 314 away from the first motor 311 is rotatably connected to the rear wall of the transfer box 3. The outer wall of the screw 314 is threadedly sleeved with a second push plate 313. A sliding interface 33 is opened inside the transfer box 3, and the second push plate 313 is slidably connected to the inside of the sliding interface 33.

[0072] In this embodiment, an oblique stopper 36 is fixed to the bottom surface of the control chassis 35, and the oblique stopper 36 is arranged on the top of the side wall of the first groove 11;

[0073] Before the tray 21 is pushed by the second pneumatic rod 32 to transfer the reagent tube 9 for puncture and separation, the positioning plate 22 on the tray 21 is fully inserted into the third groove 211 of the tray 21 under the pushing action of the inclined stop block 36, thereby ensuring the accuracy of puncturing the reagent tube 9.

[0074] The working principle and use process of the present invention:

[0075] Press first Figure 1 - Fig.12The tray 21 with the reagent tubes 9 is placed on the support seat 1. The medical staff pushes the tray 21 so that the first positioning plate 22 reaches the position of the first push plate 14, and the first pneumatic rod 13 is started. The first push plate 14 pushes the positioning plate 22 out of the tray 21. At the same time, the electric roller 73 is started to drive the positioning plate 22 to move horizontally, so that the first reagent tube 9 is stuck on the claw 77, and then the fourth pneumatic rod 78 is started to pull the second turntable 76 upward, so that the claw 77 drives the reagent tube 9 to rise, and then the third motor 72 is started to rotate the reagent tube 9 to the bottom of the first needle tube 427, and the third pneumatic rod 421 is started to pull downward. The first turntable 424 pierces the sealing plug 91 and inserts the first needle tube 427 into the reagent tube 9, starts the air pump 425 to draw the human plasma in the reagent tube 9 into the first needle tube 427, then starts the third pneumatic rod 421 to draw the first needle tube 427 out of the reagent tube 9, starts the second motor 423, rotates the first needle tube 427 to the position of the analysis device 43, the analysis device 43 is a fully automatic blood cell analyzer of model BC-5180CRP, the first needle tube 427 introduces the human plasma into the analysis device 43 for analysis, and then continues to rotate the first needle tube 427 to be cleaned and air-dried in turn;

[0076] When the first needle tube 427 is inserted downward into the reagent tube 9, the electric roller 73 continues to push the positioning plate 22 to insert the next reagent tube 9 into the claw 77, and then lifts the next reagent tube 9 upward and transfers it to the extraction component 42 position, and performs the same extraction test again. Then, the extracted reagent tube 9 rotates to the top of another tray 21. At this time, the through hole of the positioning plate 22 just moves to the bottom of the reagent tube 9, and the claw 77 is controlled to move downward. The extracted reagent tube 9 is inserted back into the positioning plate 22, and then the positioning plate 22 is pushed. The extracted reagent tube 9 and the claw 77 are separated, and the reagent tube 9 on the positioning plate 22 that has not been extracted continues to be inserted into the corresponding claw 77 until a row of reagent tubes 9 on the positioning plate 22 are extracted, tested and inserted back into the positioning plate 22. The positioning plate 22 is then driven by the electric roller 73 to be inserted into another tray 21 at the position of the first groove 11.

[0077] Then the tray 21 is pushed to continue to move, so that the next positioning plate 22 reaches the position of the first pushing plate 14, and the reagent tubes 9 are continuously transferred for extraction and testing, and the second pneumatic rod 32 is started to push another tray 21, so that the second third groove 211 of the other tray 21 is aligned with the next positioning plate 22, and the reagent tubes 9 after extraction and testing are continuously received;

[0078] Then the second pneumatic rod 32 continues to push the other tray 21, and the oblique stopper 36 pushes the positioning plate 22 to be completely inserted into the other tray 21, and the extracted reagent tube 9 reaches between the second needle tube 834 and the third needle tube 848, and the fourth motor 81 is started, and the second needle tube 834 punctures the bottom of the reagent tube 9 upwards to connect, and the third reagent tube 9 punctures the sealing plug 91 downwards to the inside of the reagent tube 9, and the human plasma in the reagent tube 9 is discharged into the waste liquid bag 5 through the second needle tube 834 and the first liquid guide tube 833, and then the water pump 844 is started again to inject the cleaning liquid in the water tank 843 into the reagent tube 9, and the residual liquid after cleaning continues to be discharged into the waste liquid bag 5, and then the medical staff throws the separated and cleaned reagent tube 9 into the recovery bag 6, until the reagent tube 9 recovered on the other tray 21 is completely cleaned, and the other tray 21 with the positioning plate 22 is pulled out to load other reagent tubes 9 to be tested;

[0079] Finally, the first motor 311 is started to drive the screw 314 to rotate, and the second push plate 313 pushes the tray 21 after the positioning plate 22 is transferred to a position close to the first groove 11, waiting to receive the positioning plate 22 and the reagent tube 9 thereon after subsequent extraction.

[0080] As a second aspect of the present invention, a method for evaluating tumor immunity is proposed, which, when implemented, comprises the following steps:

[0081] S1. Data Collection

[0082] S1-1, obtaining sample data of the questionnaire, wherein the sample data includes: lifestyle, personal medical history, family history of cancer, environmental factors and pathological data of the target patient collected by the questionnaire;

[0083] It should be noted that when the above steps are implemented, the target patient needs to fill in a lifestyle questionnaire and cancer family history information to obtain his / her informed consent, and be informed that he / she should not drink alcohol 24 hours before blood collection. Only then can the target patient be tested for health self-test, which is helpful to understand his / her health status and give a relevant score S1 based on the questionnaire answers;

[0084] S1-2, draw 2 ml of peripheral blood from the target patient, use a blood cell analyzer to detect the white blood cell count, neutrophil percentage and count, lymphocyte percentage and count, monocyte percentage and count, and determine the basic status of the target patient's blood cells, that is, according to the deviation between the test value obtained in the current test step and the normal test range, give a relevant score S2;

[0085] S1-3, based on a specific protein analyzer, the target patient's peripheral blood humoral immune protein is tested to determine its humoral immune protein;

[0086] It should be noted that, when the above steps are specifically implemented, 2 ml of human peripheral blood needs to be taken, and a specific protein analyzer is used to detect the expression of complement 3, complement 4, immunoglobulin IgE, immunoglobulin IgA, immunoglobulin IgG, and immunoglobulin IgM in the plasma. Only then can the relevant score S3 be given according to the deviation between the test value and the normal detection range;

[0087] S1-4, separate and count lymphocytes from the ex vivo blood provided by the target patient, culture lymphocytes in vitro for stimulation and cell staining, detect specific surface proteins on the surface of individual lymphocytes in the lymphocytes, and detect and calculate the number and proportion of each type of cells according to the positive or negative results of different specific surface proteins. Use flow cytometry to detect peripheral blood lymphocytes:

[0088] Specifically, the following steps are included: S1-41, 2 μL of each detection antibody is added to the flow tube, and then 100 μL of whole blood is added, mixed, and incubated at room temperature and away from light for 15 minutes; S1-42, 2.0 ml of red blood cell lysis solution is added to the same flow tube, and the tube is incubated at room temperature and away from light for 10 minutes; S1-43, centrifuged at 350 g for 5 minutes, the supernatant is discarded, the cells are washed once with PBS, centrifuged at 350 g for 5 minutes, and the supernatant is discarded; S1-44, 500 μL of PBS is added to resuspend the cells, and the cells are transferred to the flow tube and tested on the flow cytometer to obtain immune cells. Ratio and quantity; in specific implementation, 2 ml of human peripheral blood is taken, 2 μL of each detection antibody is added to the flow tube, and then 100 μL of whole blood is added, mixed, placed at room temperature, incubated in the dark for 15 minutes, and 2.0 ml of red blood cell lysis solution is added at room temperature, incubated in the dark for 10 minutes, centrifuged at 350g for 5 minutes, the supernatant is discarded, the cells are washed once with PBS, centrifuged at 350g for 5 minutes, the supernatant is discarded, 500 microliters of PBS is added to resuspend the cells, transferred to the flow tube, and detected on the flow cytometer. After obtaining the data, Flowjo Data were analyzed using V10 analysis software, Excel, and Grphpadprism8.0, and t-test was used; p < 0.05 was considered different: specifically: CD3+T lymphocytes%, CD3-CD19+B lymphocytes%, CD3-CD56+NK cells%, CD3+CD56+NK-T cells%, CD3+CD4+helper T cells%, CD3+CD8+cytotoxic T cells%, CD3+CD4+helper T cells / CD3+CD8+cytotoxic T cells, CD3 +CD4+CD25+regulatory CD4+T cells%, programmed death receptor 1%, CD3+CD4+CCR7+CD45RA+naive T cells%, CD3+CD8+CCR7+CD45RA+naive T cells%, CD3+CD4+CCR7-CD45RA-effect and memory T cells%, CD3+CD8+CCR7-CD45RA-effect and memory T cells%; Only then can the relevant score S4 be given according to the deviation of the test value from the normal test range (as shown in the table below);

[0089]

[0090] S2. Data preprocessing

[0091] S2-2. Use flow cytometry to detect the killing activity of NK cells in peripheral blood PBMC against tumor cells, and score according to the deviation between the detection value and the normal detection range; specifically include:

[0092] S2-21, mixing effector cells, i.e. PBMC cells, and target cells, i.e. tumor cells transfected with green fluorescence, into GFP-K562 cells in a certain proportion, and collecting the cells after co-culturing;

[0093] S2-22, collect cells and stain them, and use AnnexinV-APC / 7AAD to stain apoptotic and necrotic target cells;

[0094] S2-23, flow cytometry detection, in the FSC / SSC scatter plot, specifically circle all cells including effector cells and target cells, in the FITC / SSC scatter plot, divide the cells circled in Step 1 into FITC+ and FITC- cells, FITC+ refers to K562-GFP cells expressing FITC, and FITC- refers to PBMC cells with negative FITC expression, in the APC / PI scatter plot, analyze the fluorescence intensity of the APC and PI channels of the FITC+ cells in Step 2, and calculate the total proportion of K562-GFP cells with early apoptosis, mid-late apoptosis, and necrosis. Only then can the relevant score S5 be given according to the deviation between the detection value and the normal detection range;

[0095] S3. Calculate the immunity score:

[0096] According to the formula The obtained scores of each stage are weighted and the immunity score is output: where n represents the number of items tested in the immune assessment, Pi represents the item weight, and Si represents the test value of each stage of scoring, that is, Stotal = P1S1+P2S2+P3S3+P4S4+P5S5;

[0097] In the embodiment of the present invention, the total score standard of S is as follows: <60 (severe decrease in tumor immune function); 60-80 (mild decrease in tumor immune function); 80-100 (normal tumor immune function).

[0098] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, the elements defined by the sentence "comprise a ..." do not exclude the existence of other identical elements in the process, method, article or device including the elements.

[0099] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A specific protein analyzer for tumor immunity assessment, characterized in that: It includes a support seat and a reagent tube, wherein a transfer box is arranged on the top surface of the support seat, a detection mechanism is arranged on the rear wall of the transfer box, the detection mechanism and the interior of the transfer box are through-connected, a transfer assembly is arranged inside the transfer box, the top surface of the support seat is a concave structure, a sample placement assembly is arranged inside the concave structure of the support seat, a plurality of reagent tubes are inserted and placed on the top surface of the sample placement assembly, a sealing plug is sealed and connected on the top of the reagent tube, a separation mechanism is arranged on one side of the support seat, a first groove is provided inside the support seat, a second groove is provided on the bottom surface of the support seat, the separation mechanism is arranged on the top of the first groove and the second groove, a waste liquid bag is inserted in the interior of the second groove, a recovery bag is inserted in the front wall of the support seat, a pushing assembly and a second pneumatic rod are arranged on the rear wall of the transfer box, the sample placement assembly is transferred to the bottom of the separation mechanism through the cooperation of the pushing assembly and the second pneumatic rod, a control chassis is fixed on the front wall of the transfer box, and a touch screen is arranged on the front wall of the control chassis; The detection mechanism includes a detection box, an extraction component, an analysis device, a cleaning device and an air drying device. The detection box is connected to the rear wall of the transfer box. An extraction component is arranged inside the detection box. The extraction component is used to extract the reagent tube transferred by the transfer component. The extraction component is sequentially arranged with an analysis device, a cleaning device and an air drying device in the circumferential direction. The separation mechanism includes a fourth motor, a gear, a lower-through component and an upper-through component, the lower-through component is plugged into the first groove, the upper-through component is plugged into the top surface of the support seat, the fourth motor is arranged on the top surface of the support seat, the rotating end of the fourth motor is connected to a gear, the two sides of the gear are respectively meshed and connected between the upper-through component and the lower-through component, the lower-through component is used to puncture the bottom end of the reagent tube to release the residual liquid, and the upper-through component is used to stop the reagent tube from rising; The upper wear component includes a second tooth plate, a third fixed plate, a water tank, a water pump, a second lifting plate, a second liquid guide tube, a third liquid guide tube and a third needle tube. The second tooth plate is plugged into the support seat, one side of the second tooth plate is meshed with the gear, one side of the first tooth plate is fixed with the third fixed plate, the top surface of the third fixed plate is provided with a water tank, the top surface of the water tank is provided with a water pump, the side wall of the water tank is connected with the second liquid guide tube, the bottom surface of the second liquid guide tube is connected with a plurality of third liquid guide tubes, the side wall of the third fixed plate is fixed with the second lifting plate, the third liquid guide tube is plugged into the second lifting plate, and the bottom end of the third liquid guide tube is connected with the third needle tube; The sample placement assembly includes a tray and a positioning plate, a plurality of third grooves are provided on the top surface of the tray, a first limiting plate is fixed to the inner wall of the third groove, a positioning plate is inserted into the third groove, the bottom surface of the positioning plate is attached to the top surface of the first limiting plate, a through hole is provided inside the positioning plate, the through hole of the positioning plate is used for inserting the reagent tube, and a puncture groove is provided through the bottom surface of the third groove; The transfer assembly includes a second fixed plate, a third motor, a rotating shaft, a sleeve, a second rotating disk, a clamping claw and a fourth pneumatic rod. The second fixed plate is fixed to the inner wall of the transfer box. The third motor is arranged on the top surface of the second fixed plate. The rotating top of the third motor is connected to the rotating shaft. The second limiting plate is fixed to the side wall of the rotating shaft. The sleeve is sleeved on the top of the rotating shaft. The second rotating disk is fixed on the top of the sleeve. The top surface of the second rotating disk is connected to the fourth pneumatic rod. The fourth pneumatic rod is connected to the inner top surface of the transfer box. The top surface of the second rotating disk is fixed with a clamping claw, and four clamping claws are arranged in a cross distribution. The transfer assembly also includes an electric roller, which is arranged on the bottom surface of the second fixed plate. Four electric rollers are arranged in a rectangular distribution. A first pneumatic rod is arranged on the top surface of the support seat. The telescopic end of the first pneumatic rod is connected to a first push plate. The electric roller and the first push plate are both used to push the positioning plate to translate.

2. The specific protein analyzer for tumor immunity assessment according to claim 1, characterized in that: The down-penetrating assembly includes a first tooth plate, a first lifting plate, a first liquid guiding tube and a second needle tube. The first tooth plate is inserted into the support seat, one side of the first tooth plate is meshed with the gear, the first lifting plate is fixed to the side wall of the first tooth plate, the first lifting plate is arranged in the first groove, a plurality of first liquid guiding tubes are inserted in the first lifting plate, the top end of the first liquid guiding tube is connected to the second needle tube, and the bottom end of the first liquid guiding tube is arranged in the waste liquid bag.

3. The specific protein analyzer for tumor immunity assessment according to claim 2, characterized in that: The extraction component includes a third pneumatic rod, a third push plate, a second motor, a first turntable, an air pump, a guide tube and a first needle tube. The third pneumatic rod is arranged inside the detection box. The telescopic top end of the third pneumatic rod is connected to the third push plate. The top surface of the third push plate is provided with the second motor. The rotating top end of the second motor is connected to the first turntable. The top surface of the first turntable is provided with an air pump. The side wall of the air pump is connected to the guide tube. Four guide tubes are arranged in a cross distribution. The bottom surface of the guide tube is connected with the first needle tube.

4. The specific protein analyzer for tumor immunity assessment according to claim 3, characterized in that: The pushing assembly includes a first motor, a first fixed plate, a second push plate and a screw. The first fixed plate is fixed to the rear wall of the transfer box. The first fixed plate is provided with a first motor on the side wall. The rotating end of the first motor is connected to the screw. The end of the screw away from the first motor is rotatably connected to the rear wall of the transfer box. The outer wall of the screw is threadedly sleeved with a second push plate. A sliding interface is provided inside the transfer box, and the second push plate is slidably connected to the inside of the sliding interface.

5. The specific protein analyzer for tumor immunity assessment according to claim 4, characterized in that: An oblique stopper is fixed on the bottom surface of the control chassis, and the oblique stopper is arranged on the top of the side wall of the first groove.

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