A leukemia tumor cell detection kit

Abstract

The present application relates to the technical field of biological detection, and particularly relates to a leukemia tumor cell detection kit. The leukemia tumor cell detection kit provided by the present application comprises a monoclonal antibody and a compound buffer; the monoclonal antibody comprises CD71 antibody, CD56 antibody, CD38 antibody, CD19 antibody, CD10 antibody, CD64 antibody, CD15 antibody, CD13 antibody, CD7 antibody and CD33 antibody; the monoclonal antibody is marked with a detection marker; the compound buffer comprises glycine, leucine, potassium dihydrogen phosphate, sodium hydroxide and deionized water; the pH value of the compound buffer is 7.2-7.4. The present application constructs a new compound buffer, which can better stabilize the antibody and other components in the leukemia tumor cell detection kit, obviously prolongs the effective period of detection precision and detection efficiency, and greatly saves the detection cost.

Description

Technical Field

[0001] This invention relates to the field of biological detection technology, and in particular to a leukemia tumor cell detection kit. Background Technology

[0002] Leukemia, a common hematologic malignancy, requires efficient and accurate detection as a crucial step in its prevention and treatment. Leukemia tumor cell detection kits, as important tools for diagnosis, subtyping, and monitoring of leukemia, involve fusion gene detection, gene mutation analysis, and immunophenotyping. Clinically available kits include the BCR-ABL fusion gene detection kit, the JAK2-V617F gene mutation detection kit, and leukemia cell immunophenotyping kits.

[0003] Among them, the leukemia cell immunophenotyping assay kit contains a variety of antibodies. By detecting cell surface differentiation antigens, it can classify leukemia cells, distinguish different types of leukemia, and provide key diagnostic and monitoring information for clinical use.

[0004] Currently, regarding leukemia tumor cell detection kits, researchers are continuously improving the detection accuracy and efficiency of these kits through new antibodies and other methods. However, how to better maintain the detection accuracy and efficiency of leukemia tumor cell detection kits is equally important for this field. Summary of the Invention

[0005] In view of this, the present invention provides a leukemia tumor cell detection kit, which can better maintain detection accuracy and detection efficiency.

[0006] This invention provides a leukemia tumor cell detection kit, comprising monoclonal antibodies and a complex buffer; the monoclonal antibodies include CD71 antibody, CD56 antibody, CD38 antibody, CD19 antibody, CD10 antibody, CD64 antibody, CD15 antibody, CD13 antibody, CD7 antibody, and CD33 antibody; all monoclonal antibodies are labeled with detection markers; the complex buffer includes glycine, leucine, potassium dihydrogen phosphate, sodium hydroxide, and deionized water; the pH of the complex buffer is 7.2-7.4.

[0007] Preferably, each of the monoclonal antibodies is a mouse-derived or human-derived monoclonal antibody.

[0008] Preferably, each of the monoclonal antibodies is labeled with at least one of FITC, PE-Cy7, Alexa Fluor 488, PerCP-cy7, APC, APC-Alexa Fluor700, PB, ECD, PE and PerCP-cy5.5.

[0009] Preferably, the CD71 antibody is labeled with FITC; the CD56 antibody is labeled with PE-Cy7; the CD38 antibody is labeled with Alexa Fluor 488; the CD19 antibody is labeled with PerCP-cy7; the CD10 antibody is labeled with APC; the CD64 antibody is labeled with APC-Alexa Fluor700; the CD15 antibody is labeled with PB; the CD13 antibody is labeled with ECD; the CD7 antibody is labeled with PE; and the CD33 antibody is labeled with PerCP-cy5.5.

[0010] Preferably, in the composite buffer solution, the mass ratio of glycine to leucine is 0.356~0.472: 0.327~0.521.

[0011] Preferably, the mass ratio of glycine to leucine in the composite buffer solution is 0.411:0.431.

[0012] Preferably, in the composite buffer solution, the mass ratio of glycine to potassium dihydrogen phosphate is 0.356~0.472: 1~3.

[0013] Preferably, the mass ratio of glycine to potassium dihydrogen phosphate in the composite buffer solution is 0.411:2.

[0014] Preferably, the molar ratio of potassium dihydrogen phosphate to sodium hydroxide is 8~12:8~10.

[0015] Preferably, the molar ratio of potassium dihydrogen phosphate to sodium hydroxide is 10:9.

[0016] Preferably, the mass ratio of potassium dihydrogen phosphate to deionized water is 13~14:2800~3000.

[0017] Preferably, the mass ratio of potassium dihydrogen phosphate to deionized water is 13.6:2850.

[0018] This invention addresses the aforementioned antibodies by constructing a specific composite buffer solution, thereby endowing the leukemia tumor cell detection kit with better overall performance: on the one hand, it can maintain the stability of the system's pH value, ensuring the stability of the antibody's function; on the other hand, it can provide a more suitable ionic environment for the leukemia tumor cell detection kit; crucially, it can improve the activity and stability of the aforementioned antibodies, thus providing more accurate detection results for subsequent tests.

[0019] Preferably, the leukemia tumor cell detection kit further includes eukaryotic initiation factor 4E (eIF4E).

[0020] Preferably, the leukemia tumor cell detection kit further includes red blood cell lysis buffer.

[0021] Preferably, the erythrocyte lysis buffer comprises NH4Cl, NaHCO3, disodium ethylenediaminetetraacetate (EDTA-2Na), and water; the mass ratio of NH4Cl to NaHCO3 is 8~12:1; the mass ratio of NaHCO3 to disodium ethylenediaminetetraacetate is 1~3:1; the mass ratio of NH4Cl to water is 1:11~13; and the pH value of the erythrocyte lysis buffer is 7.2~7.4.

[0022] Preferably, the mass ratio of NH4Cl to NaHCO3 is 9:1; the mass ratio of NaHCO3 to disodium ethylenediaminetetraacetate is 2:1; the mass ratio of NH4Cl to water is 1:12; and the pH value of the erythrocyte lysis buffer is 7.3.

[0023] This invention provides a leukemia tumor cell detection kit. Through long-term clinical research, this invention has developed a novel composite buffer solution that better stabilizes the antibodies and other components in the leukemia tumor cell detection kit, significantly extending its detection accuracy and efficiency, greatly reducing detection costs, and providing greater convenience in production, transportation, and use, resulting in significant social and economic benefits. The leukemia tumor cell detection kit provided by this invention can be used to detect acute myeloid leukemia, acute myeloid leukemia M3, myelodysplastic syndrome (MDS), normal bone marrow with reactive B-cell hyperplasia, acute myeloid leukemia M5, marginal zone lymphoma (MZL), and Castleman 10 disease. Attached Figure Description

[0024] To more clearly illustrate the technical solutions of this invention, the accompanying drawings used in the embodiments of this invention or in the prior art are briefly described below. For those skilled in the art, other drawings can be derived from the following drawings without creative effort, and all such drawings are within the protection scope of this invention.

[0025] Figure 1 This is a comparison chart of the detection efficiency of the embodiments and comparative examples of the present invention; wherein, the relative detection time refers to the value of the detection time of other embodiments (embodiments 7 or 9) or comparative examples (comparative examples 1 to 9) relative to the detection time of embodiment 5, with the detection time of embodiment 5 recorded as 100. Detailed Implementation

[0026] This invention provides a leukemia tumor cell detection kit, comprising monoclonal antibodies and a complex buffer; the monoclonal antibodies include CD71 antibody, CD56 antibody, CD38 antibody, CD19 antibody, CD10 antibody, CD64 antibody, CD15 antibody, CD13 antibody, CD7 antibody, and CD33 antibody; all monoclonal antibodies are labeled with detection markers; the complex buffer includes glycine, leucine, potassium dihydrogen phosphate, sodium hydroxide, and deionized water; the pH of the complex buffer is 7.2-7.4.

[0027] In specific embodiments of the present invention, each of the monoclonal antibodies is preferably a mouse or human monoclonal antibody.

[0028] In specific embodiments of the present invention, each of the monoclonal antibody labels is at least one of FITC, PE-Cy7, Alexa Fluor488, PerCP-cy7, APC, APC-Alexa Fluor700, PB, ECD, PE and PerCP-cy5.5.

[0029] In specific embodiments of the present invention, the CD71 antibody is preferably labeled with FITC; the CD56 antibody is preferably labeled with PE-Cy7; the CD38 antibody is preferably labeled with Alexa Fluor 488; the CD19 antibody is preferably labeled with PerCP-cy7; the CD10 antibody is preferably labeled with APC; the CD64 antibody is preferably labeled with APC-Alexa Fluor700; the CD15 antibody is preferably labeled with PB; the CD13 antibody is preferably labeled with ECD; the CD7 antibody is preferably labeled with PE; and the CD33 antibody is preferably labeled with PerCP-cy5.5.

[0030] In a specific embodiment of the present invention, the mass ratio of glycine to leucine in the composite buffer solution is preferably 0.356~0.472:0.327~0.521, more preferably 0.411:0.431.

[0031] In a specific embodiment of the present invention, the mass ratio of glycine to potassium dihydrogen phosphate is based on the pH value of the composite buffer solution being 7.2 to 7.4, preferably 0.356 to 0.472:1 to 3, and more preferably 0.411:2.

[0032] In a specific embodiment of the present invention, the molar ratio of potassium dihydrogen phosphate to sodium hydroxide is preferably 8~12:8~10, and more preferably 10:9.

[0033] In a specific embodiment of the present invention, the mass ratio of potassium dihydrogen phosphate to deionized water is preferably 13~14:2800~3000, more preferably 13.6:2850.

[0034] This invention addresses the aforementioned antibodies by constructing a specific composite buffer solution, thereby endowing the leukemia tumor cell detection kit with better overall performance: on the one hand, it can maintain the stability of the system's pH value, ensuring the stability of the antibody's function; on the other hand, it can provide a more suitable ionic environment for the leukemia tumor cell detection kit; crucially, it can improve the activity and stability of the aforementioned antibodies, thus providing more accurate detection results for subsequent tests.

[0035] In a specific embodiment of the present invention, the leukemia tumor cell detection kit preferably further includes eukaryotic initiation factor 4E (eIF4E).

[0036] In a specific embodiment of the present invention, the leukemia tumor cell detection kit preferably further includes erythrocyte lysis buffer; the erythrocyte lysis buffer preferably includes NH4Cl, NaHCO3, disodium ethylenediaminetetraacetate (EDTA-2Na), and water; the mass ratio of NH4Cl to NaHCO3 is preferably 8~12:1, more preferably 9:1; the mass ratio of NaHCO3 to disodium ethylenediaminetetraacetate is preferably 1~3:1, more preferably 2:1; the mass ratio of NH4Cl to water is preferably 1:11~13, more preferably 1:12; the pH value of the erythrocyte lysis buffer is preferably 7.2~7.4, more preferably 7.3.

[0037] The leukemia tumor cell detection kit provided by this invention can be used to detect acute myeloid leukemia, acute myeloid leukemia M3, myelodysplastic syndrome (MDS), normal bone marrow with reactive B-cell hyperplasia, acute myeloid leukemia M5, marginal zone lymphoma (MZL), and Castleman 10 disease.

[0038] To further illustrate the present invention, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings and embodiments.

[0039] Example 1: In this embodiment, the monoclonal antibodies used were fluorescently labeled monoclonal antibodies. Samples of each monoclonal antibody were taken, serially diluted, and detected using flow cytometry. The specific dosage of each antibody was determined by titration, as follows: Fluorescently labeled monoclonal antibody CD71-FITC (5 μL); fluorescently labeled monoclonal antibody CD56-PE-Cy7 (5 μL); fluorescently labeled monoclonal antibody CD38-Alexa Fluor 488 (1 μL); fluorescently labeled monoclonal antibody CD19-PerCP-cy7 (2 μL); fluorescently labeled monoclonal antibody CD10-APC (1 μL); fluorescently labeled monoclonal antibody CD64-APC-Alexa Fluor 700 (2 μL); fluorescently labeled monoclonal antibody CD15-PB (1 μL); fluorescently labeled monoclonal antibody CD13-ECD (1 μL); fluorescently labeled monoclonal antibody CD7-PE (2 μL); fluorescently labeled monoclonal antibody CD33-PerCP-cy5.5 (5 μL).

[0040] The above fluorescently labeled monoclonal antibodies were mixed and packaged into the same flow cytometry tube to obtain a leukemia tumor cell detection kit.

[0041] Example 2: The specimen was prepared in this embodiment: The specific steps are as follows: Peripheral blood anticoagulated with EDTA was used as a sample, and the cell density was adjusted to 1-5 × 10⁻⁶. 6 / ml, then filter the sample using a 200-mesh sieve to remove clumps and impurities, ensuring that the individual cells are in suspension, and store at 2~8℃ to obtain the specimen.

[0042] Example 3: This embodiment prepared a complex buffer solution, and the specific steps are as follows: Glycine and leucine were mixed in a mass ratio of 0.472:0.521; potassium dihydrogen phosphate and sodium hydroxide were mixed in a molar ratio of 8:10; and the two mixtures prepared above were mixed with deionized water in a mass ratio of 13.6:2850, with a pH of 7.4, to obtain the composite buffer solution.

[0043] Example 4: Take 50 μL of the sample prepared in Example 2, vortex mix it, and incubate it at 25°C in the dark for 15 minutes.

[0044] Weigh 80.1g of NH4Cl, 8.4g of NaHCO3 and 3.7g of EDTA-2Na, add water to 900mL, mix well, adjust the pH of the solution to 7.4, add water to 1L to obtain 10× red blood cell lysis buffer.

[0045] Using the composite buffer prepared in Example 3, the 10× erythrocyte lysis buffer was diluted to 1× erythrocyte lysis buffer. Then, 2 mL of 1× erythrocyte lysis buffer was added to the incubated flow cytometry tube, and the mixture was vortexed and incubated at 25°C in the dark for 10 minutes.

[0046] Centrifuge the flow cytometry tubes at 1500 rpm for 5 minutes, discard the supernatant, resuspend in 200 μL of compound buffer, and detect using a four-color flow cytometer.

[0047] Example 5: Blood samples were collected from 30 leukemia patients and 30 healthy individuals (2 mL each). Serum was collected by centrifugation and analyzed according to the steps in Examples 1-4. The leukemia tumor cell detection kit was used on the same day it was prepared. The results are shown in Table 1 and... Figure 1 As shown.

[0048] Table 1. Detection results of Example 5:

[0049] According to Table 1 and Figure 1 As can be seen, the leukemia tumor cell detection kit provided by this invention can achieve high detection accuracy, and the detection process is convenient, fast, simple and efficient.

[0050] Example 6: This embodiment prepared a complex buffer solution, and the specific steps are as follows: Glycine and leucine were mixed in a mass ratio of 0.356:0.327; potassium dihydrogen phosphate and sodium hydroxide were mixed in a molar ratio of 12:10; and the two mixtures prepared above were mixed with deionized water in a mass ratio of 13.6:2850, with a pH of 7.2, to obtain the composite buffer solution.

[0051] Example 7: Blood samples were collected from 30 leukemia patients and 30 healthy individuals (2 mL each). Serum was collected by centrifugation and analyzed according to the steps in Examples 1-2, 6, and 4. The leukemia tumor cell detection kit was opened and stored (in a simulated environment to accelerate detection stability). The kit was used on day 15 after preparation. Results are shown in Table 2. Figure 1 As shown.

[0052] Table 2 Detection results of Example 7:

[0053] According to Table 2 and Figure 1It can be seen that the leukemia tumor cell detection kit provided by the present invention can still maintain high detection accuracy and high detection efficiency after being opened and stored for 15 days. This indicates that the composite buffer provided by the present invention can enable the leukemia tumor cell detection kit to better maintain detection accuracy and detection efficiency.

[0054] Example 8: This embodiment prepared a complex buffer solution, and the specific steps are as follows: Glycine and leucine were mixed in a mass ratio of 0.411:0.431; potassium dihydrogen phosphate and sodium hydroxide were mixed in a molar ratio of 10:9; and the two mixtures prepared above were mixed with deionized water in a mass ratio of 13.6:2850, with a pH of 7.2, to obtain the composite buffer solution.

[0055] Example 9: Blood samples were collected from 30 leukemia patients and 30 healthy individuals (2 mL each). Serum was collected by centrifugation and analyzed according to the steps in Examples 1-2, 8, and 4. The leukemia tumor cell detection kit was opened and stored (in a simulated environment to accelerate detection stability). The kit was used 45 days after preparation. Results are shown in Table 3. Figure 1 As shown.

[0056] Table 3 Detection results of Example 9:

[0057] According to Table 3 and Figure 1 It can be seen that the leukemia tumor cell detection kit provided by the present invention can still maintain high detection accuracy and high efficiency after being opened and stored for 45 days. This indicates that the composite buffer provided by the present invention can better maintain the detection accuracy and efficiency of the leukemia tumor cell detection kit.

[0058] Comparative Example 1: This comparative example used commercially available PBS phosphate buffer (pH 7.2). 2 mL of blood was collected from 30 leukemia patients and 30 healthy individuals. Serum was collected by centrifugation and analyzed according to Examples 1-2, this comparative example, and Example 4. The leukemia tumor cell detection kit was opened and stored (in a simulated environment to accelerate detection stability). The kit was used on the 5th day after preparation. Results are shown in Table 4. Figure 1 As shown.

[0059] Table 4. Detection results of Comparative Example 1:

[0060] According to Table 4 and Figure 1 As can be seen, the leukemia tumor cell detection kit provided in this comparative example can still maintain high detection accuracy and the detection process remains efficient even after being opened and stored for 5 days.

[0061] Comparative Example 2: This comparative example used commercially available PBS phosphate buffer (pH 7.2). 2 mL of blood was collected from 30 leukemia patients and 30 healthy individuals. Serum was collected by centrifugation and analyzed according to Examples 1-2, this comparative example, and Example 4. The leukemia tumor cell detection kit was opened and stored (in a simulated environment to accelerate detection stability). The kit was used on day 15 after preparation. Results are shown in Table 5. Figure 1 As shown.

[0062] Table 5. Detection results of Comparative Example 2:

[0063] According to Table 5 and Figure 1 It can be seen that the leukemia tumor cell detection kit provided in this comparative example showed a decrease in detection accuracy after being opened and stored for 15 days, but the detection process was relatively efficient.

[0064] Comparative Example 3: This comparative example used commercially available PBS phosphate buffer (pH 7.2). 2 mL of blood was collected from 30 leukemia patients and 30 healthy individuals. Serum was collected by centrifugation and analyzed according to Examples 1-2, this comparative example, and Example 4. The leukemia tumor cell detection kit was opened and stored (in a simulated environment to accelerate detection stability). The kit was used on day 45 after preparation. Results are shown in Table 6. Figure 1 As shown.

[0065] Table 6. Detection results of Comparative Example 3:

[0066] According to Table 6 and Figure 1 It can be seen that the leukemia tumor cell detection kit provided in this comparative example showed a significant decrease in detection accuracy and a longer detection time after being opened and stored for 45 days.

[0067] Comparative Example 4: This comparative example uses the preparation method of Example 3, the only difference being that glycine is replaced with an equal mass of leucine. 2 mL of blood was taken from 30 leukemia patients and 30 healthy individuals. Serum was collected by centrifugation and analyzed according to Examples 1-2, this comparative example, and Example 4. The leukemia tumor cell detection kit was opened and stored (in a simulated environment to accelerate detection stability). The kit was used on the 5th day after preparation, and the results are shown in Table 7. Figure 1 As shown.

[0068] Table 7. Detection results of Comparative Example 4:

[0069] According to Table 7 and Figure 1 As can be seen, the leukemia tumor cell detection kit provided in this comparative example can still maintain high detection accuracy and high efficiency even after being opened and stored for 5 days.

[0070] Comparative Example 5: This comparative example uses the preparation method of Example 3, the only difference being that glycine is replaced with an equal mass of leucine. 2 mL of blood was taken from 30 leukemia patients and 30 healthy individuals, and the serum was collected by centrifugation. The serum was tested according to Examples 1-2, this comparative example, and Example 4. The leukemia tumor cell detection kit was opened and placed in a simulated environment to accelerate detection stability. It was used on the 15th day after preparation, and the results are shown in Table 8. Figure 1 As shown.

[0071] Table 8. Detection results of Comparative Example 5:

[0072] According to Table 8 and Figure 1 As can be seen, the leukemia tumor cell detection kit provided in this comparative example maintains high detection accuracy and high efficiency even after being opened and stored for 15 days.

[0073] Comparative Example 6: This comparative example uses the preparation method of Example 3, the only difference being that glycine is replaced with an equal mass of leucine. 2 mL of blood was taken from 30 leukemia patients and 30 healthy individuals, and the serum was collected by centrifugation. The serum was tested according to Examples 1-2, this comparative example, and Example 4. The leukemia tumor cell detection kit was opened and stored (in a simulated environment to accelerate detection stability). The kit was used 45 days after preparation, and the results are shown in Table 9. Figure 1 As shown.

[0074] Table 9. Test results of Comparative Example 6:

[0075] According to Table 9 and Figure 1 It can be seen that the leukemia tumor cell detection kit provided in this comparative example has significantly insufficient detection accuracy after being opened and stored for 45 days, and the detection process requires a relatively long time.

[0076] Comparative Example 7: This comparative example uses the preparation method of Example 3, the only difference being that leucine is replaced with an equal mass of glycine. 2 mL of blood was taken from 30 leukemia patients and 30 healthy individuals. Serum was collected by centrifugation and analyzed according to Examples 1-2, this comparative example, and Example 4. The leukemia tumor cell detection kit was opened and stored (in a simulated environment to accelerate detection stability). The kit was used on the 5th day after preparation, and the results are shown in Table 10. Figure 1 As shown.

[0077] Table 10: Test results of Comparative Example 7

[0078] According to Table 10 and Figure 1 As can be seen, the leukemia tumor cell detection kit provided in this comparative example maintains good detection accuracy after being opened and stored for 5 days, and the detection process is efficient and simple.

[0079] Comparative Example 8: This comparative example uses the preparation method of Example 3, the only difference being that leucine is replaced with an equal mass of glycine. 2 mL of blood was taken from 30 leukemia patients and 30 healthy individuals. Serum was collected by centrifugation and analyzed according to Examples 1-2, this comparative example, and Example 4. The leukemia tumor cell detection kit was opened and stored (in a simulated environment to accelerate detection stability). The kit was used on day 15 after preparation, and the results are shown in Table 11 and... Figure 1 As shown.

[0080] Table 11 Detection results of Comparative Example 8:

[0081] According to Table 11 and Figure 1 As can be seen, the leukemia tumor cell detection kit provided in this comparative example maintains good detection accuracy after being opened and stored for 15 days, and the detection process is efficient and simple.

[0082] Comparative Example 9: This comparative example uses the preparation method of Example 3, the only difference being that leucine is replaced with an equal mass of glycine. 2 mL of blood was taken from 30 leukemia patients and 30 healthy individuals, and the serum was collected by centrifugation. The serum was tested according to Examples 1-2, this comparative example, and Example 4. The leukemia tumor cell detection kit was opened and stored (in a simulated environment to accelerate detection stability). The kit was used 45 days after preparation, and the results are shown in Table 12 and... Figure 1 As shown.

[0083] Table 12 Detection results of Comparative Example 9:

[0084] According to Table 12 and Figure 1 It can be seen that the leukemia tumor cell detection kit provided in this comparative example showed a decrease in detection accuracy and a decrease in detection efficiency after being opened and stored for 45 days.

[0085] As can be seen from the above examples and comparative examples, the leukemia tumor cell detection kit provided by the present invention significantly improves the stability of components such as antibodies through the construction of composite buffer. The detection accuracy and efficiency remain basically unchanged within 45 days, which is significantly better than the existing level (generally within 30 days after opening). It provides a new method for reducing the cost of leukemia tumor cell detection and has good application prospects.

[0086] The embodiments of the present invention have been described above; however, these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. All other embodiments obtained by those skilled in the art based on the above embodiments of the present invention without inventive effort are within the protection scope of the present invention.

Claims

1. A leukemia tumor cell detection kit, characterized in that, Includes monoclonal antibodies and complex buffers; The monoclonal antibodies include CD71 antibody, CD56 antibody, CD38 antibody, CD19 antibody, CD10 antibody, CD64 antibody, CD15 antibody, CD13 antibody, CD7 antibody, and CD33 antibody; All monoclonal antibodies were labeled with detection markers; The composite buffer solution comprises glycine, leucine, potassium dihydrogen phosphate, sodium hydroxide, and deionized water; The pH value of the composite buffer solution is 7.2~7.

4.

2. The leukemia tumor cell detection kit according to claim 1, characterized in that, Each of the monoclonal antibodies is labeled with at least one of FITC, PE-Cy7, Alexa Fluor 488, PerCP-cy7, APC, APC-Alexa Fluor700, PB, ECD, PE and PerCP-cy5.

5.

3. The leukemia tumor cell detection kit according to claim 2, characterized in that, The CD71 antibody is labeled with FITC; The CD56 antibody is labeled with fluorescein PE-Cy7; The CD38 antibody is labeled with Alexa Fluor 488; The CD19 antibody is labeled with the fluorescein PerCP-cy7; The CD10 antibody is labeled with fluorescein APC; The CD64 antibody is labeled with fluorophore APC-Alexa Fluor700; The CD15 antibody is labeled with fluorescein PB; The CD13 antibody is labeled with fluorescein ECD; The CD7 antibody is labeled with fluorescein PE; The CD33 antibody was labeled with the fluorescein PerCP-cy5.

5.

4. The leukemia tumor cell detection kit according to claim 1, characterized in that, In the composite buffer solution, the mass ratio of glycine to leucine is 0.356~0.472: 0.327~0.

521.

5. The leukemia tumor cell detection kit according to claim 1, characterized in that, In the composite buffer solution, the mass ratio of glycine to potassium dihydrogen phosphate is 0.356~0.472: 1~3.

6. The leukemia tumor cell detection kit according to claim 1, characterized in that, The molar ratio of potassium dihydrogen phosphate to sodium hydroxide is 8~12:8~10.

7. The leukemia tumor cell detection kit according to claim 1, characterized in that, The mass ratio of potassium dihydrogen phosphate to deionized water is 13~14:2800~3000.

8. The leukemia tumor cell detection kit according to claim 1, characterized in that, The leukemia tumor cell detection kit also includes eukaryotic initiation factor 4E.

9. The leukemia tumor cell detection kit according to claim 1, characterized in that, The leukemia tumor cell detection kit also includes red blood cell lysis buffer.

10. The leukemia tumor cell detection kit according to claim 9, characterized in that, The erythrocyte lysis buffer comprises NH4Cl, NaHCO3, disodium ethylenediaminetetraacetate, and water; the mass ratio of NH4Cl to NaHCO3 is 8-12:1; the mass ratio of NaHCO3 to disodium ethylenediaminetetraacetate is 1-3:1; the mass ratio of NH4Cl to water is 1:11-13; and the pH of the erythrocyte lysis buffer is 7.2-7.4.

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