Flow type detection panel for primary screening of tumors in blood system and application of flow type detection panel

Abstract

The invention discloses a flow type detection panel for primary screening of tumors in a blood system and application of the flow type detection panel. The flow type detection panel comprises a T tube, a B tube and an M tube, the T tube comprises 10 channels, and is composed of the following antibodies: CD5, CD7, CD45, TRBC1, CD30 / 117, CD56, CD4, CD2, CD3 and CD8; the B tube comprises nine channels, and the B tube is composed of the following antibodies: CD5, CD56, CD45, cKappa, cLambda, CD38, CD10, CD19 and CD20; the M tube comprises 10 channels and is composed of the following antibodies: CD13, CD11b, CD45, CD14, CD64, CD33, CD117, CD34, HLA DR (human leukocyte antigen DR) and CD38. The 10-color multi-parameter flow detection Panel has multiple channels and accurate antibodies, and can accurately reflect the proportion and expression condition of each subgroup, so that the accuracy of molecular detection of cross-line expression is improved, and the diagnosis of rare diseases is further improved.

Description

Technical Field

[0001] The present invention belongs to the field of biological detection technology, and in particular relates to a flow detection panel for primary screening of hematological tumors and its application. Background Art

[0002] Hematologic diseases refer to conditions affecting the blood, bone marrow, lymphatic system, and coagulation function, encompassing a wide range of categories, including anemia, leukemia, lymphoma, and myelodysplastic syndrome. In recent years, the China Cancer Center (NCC) has released data on the incidence and mortality of new cancers in China in 2022, noting that hematologic malignancies have received increasing attention year by year. Leukemia / lymphoma, a disease characterized by the clonal proliferation of hematopoietic cells, is one of the most common hematologic malignancies.

[0003] Currently, the diagnosis of hematologic malignancies primarily relies on morphology, immunology, cytogenetics, and molecular biology. With advances in medicine, MICM technology has been rapidly updated, providing crucial diagnostic evidence for the WHO's hematologic malignancy diagnosis. Immunology, as a crucial component of hematologic malignancy diagnosis, plays a crucial role in disease diagnosis and prognosis monitoring. Currently, multicolor flow cytometry, the primary diagnostic technique for immunological disease monitoring, has transitioned from four-color flow cytometry to multiparameter flow cytometry, with throughput rates gradually increasing. According to literature reports, 10-color flow cytometry is the most widely used in the diagnosis of hematologic malignancies due to its accuracy, efficiency, and stability. However, the performance of 10-color flow cytometry in the diagnosis of hematologic malignancies varies widely.

[0004] This invention introduces a leukemia / lymphoma initial screening test panel designed based on a 10-color multi-parameter flow cytometer to address the initial screening needs of clinical hematological tumor immunology testing and provide a basis for the next step of disease subtype classification. Summary of the Invention

[0005] The purpose of the present invention is to provide a genetic detection panel for the initial screening of hematological tumors and its application. Due to its multiple channels and precise antibodies, it can accurately reflect the proportion and expression of each subpopulation, thereby improving the accuracy of molecular detection of cross-lineage expression and further improving the diagnosis of rare diseases.

[0006] In order to achieve the above objectives, this application adopts the following technical solutions:

[0007] In a first aspect, the present invention provides a flow cytometry panel for the initial screening of hematological tumors, comprising a T tube, a B tube, and an M tube; the T tube comprises 10 channels composed of the following antibodies: CD5, CD7, CD45, TRBC1, CD30 / 117, CD56, CD4, CD2, CD3, and CD8;

[0008] The B tube includes 9 channels composed of the following antibodies: CD5, CD56, CD45, cKappa, cLambda, CD38, CD10, CD19, and CD20;

[0009] The M tube includes 10 channels composed of the following antibodies: CD13, CD11b, CD45, CD14, CD64, CD33, CD117, CD34, HLA DR and CD38.

[0010] In the above technical solution, the blood system tumors include leukemia, lymphoma, multiple myeloma, paroxysmal nocturnal hemoglobinuria, chronic bone proliferative tumors, myeloproliferative tumors and megaloblastic anemia.

[0011] In a second aspect, the present invention provides the use of the above-mentioned flow detection panel in preparing a primary screening device for hematological tumors.

[0012] In a third aspect, the present invention provides the use of the above-mentioned flow cytometry panel in the preparation of a primary screening reagent for hematological tumors.

[0013] In a fourth aspect, the present invention provides a flow cytometry kit for the initial screening of hematological tumors, the kit containing antibodies respectively labeled with different fluorescein, the antibodies being divided into three groups, the first group consisting of the following antibodies: CD5, CD7, CD45, TRBC1, CD30 / 117, CD56, CD4, CD2, CD3, and CD8;

[0014] The second panel consisted of the following antibodies: CD5, CD56, CD45, cKappa, cLambda, CD38, CD10, CD19, and CD20;

[0015] The third panel consisted of the following antibodies: CD13, CD11b, CD45, CD14, CD64, CD33, CD117, CD34, HLA DR, and CD38;

[0016] Each antibody in each set is packaged separately.

[0017] In a fifth aspect, the present invention provides a detection reagent for the initial screening of hematological tumors, the detection reagent comprising antibodies respectively labeled with different fluorescein, the antibodies being divided into three groups, the first group consisting of the following antibodies: CD5, CD7, CD45, TRBC1, CD30 / 117, CD56, CD4, CD2, CD3, and CD8;

[0018] The second panel consisted of the following antibodies: CD5, CD56, CD45, cKappa, cLambda, CD38, CD10, CD19, and CD20;

[0019] The third panel consisted of the following antibodies: CD13, CD11b, CD45, CD14, CD64, CD33, CD117, CD34, HLA DR, and CD38;

[0020] Each antibody in each set is packaged separately.

[0021] In a sixth aspect, the present invention provides a system for primary screening of hematological tumors, comprising a detection portion and an analysis portion; the detection portion comprises the above-mentioned detection reagent;

[0022] The analyzing part is used to analyze the detection results of the detecting part.

[0023] The beneficial effects of the present invention lie in the following: the 10-color multiparameter flow cytometry panel designed in this invention is designed to improve the sensitivity of the initial diagnosis of acute leukemia (AL) subtype classification, non-Hodgkin's lymphoma (NHL), paronychia nodosum (PNH), and non-hematopoietic tumors in the blood system. Several years ago, leukemia / lymphoma detection was primarily performed using traditional 4-color and 8-color panels, which often only met the most basic clinical diagnostic needs. In recent years, a large number of new CD molecules have been discovered and applied to flow cytometry for hematological diseases, prompting the continuous upgrading of flow cytometry technology. Traditional 4-color and 8-color panels have reached a bottleneck in the diagnostic criteria for hematological diseases. 10-color multiparameter flow cytometry, by comparison, enables high-dimensional data acquisition, simultaneous detection of multiple targets, and reduced batch variability. Furthermore, with increasing sample sizes and low cell counts in individual samples, 10-color multiparameter flow cytometry can improve detection efficiency, reduce reagent waste, and reduce labor costs. Furthermore, 10-color multi-parameter flow cytometry, with its multiple channels and precise antibodies, can accurately reflect the proportion and expression of each subpopulation, thereby improving the accuracy of molecular detection of cross-lineage expression and further improving the diagnosis of rare diseases. With the continuous development of multi-parameter flow cytometry, more and more flow cytometers with multiple channels have been introduced, and the complexity of fluorescence compensation, the accuracy of instrument data, and the specialization of data analysis have gradually increased. However, these will place a heavy burden on clinical work and thus fail to provide a high-quality detection basis for the clinic. However, 10-color flow cytometry, through its multi-parameter simultaneous detection, high-resolution clustering, and data integration capabilities, can not only meet the needs of clinical diagnosis but also provide favorable guarantees for subsequent precise diagnosis and treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 It is a flow chart of the detection method of the present invention.

[0025] Figure 2 This is a two-dimensional dot plot (T tube) of flow cytometry according to Example 1 of the present invention.

[0026] Figure 3 This is a two-dimensional dot plot (B tube) of flow cytometry according to Example 1 of the present invention.

[0027] Figure 4 This is a two-dimensional dot plot (M tube) of flow cytometry according to Example 1 of the present invention. DETAILED DESCRIPTION

[0028] In order to better illustrate the purpose, technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments. The present invention can be implemented in many different forms and should not be construed as limited to the embodiments set forth herein. On the contrary, these embodiments are provided so that this disclosure will be thorough and complete and will fully convey the concept of the present invention to those skilled in the art. The present invention will be limited only by the claims.

[0029] The present invention provides a gene detection panel for the initial screening of hematological tumors and its application, which accurately and efficiently utilizes multi-parameter flow cytometry to initially screen hematological tumors.

[0030] 1. Panel formulation principles

[0031] Based on the consensus on flow cytometry, the present invention combines the two diagnostic criteria of EGIL (1995 edition) and WHO (2008 / 2016 edition) to propose for the first time a 10-color multi-parameter flow cytometry protocol for the detection of leukemia / lymphoma. When formulating the panel, we strictly select antibodies that have been verified by multiple experiments and have a good reputation to ensure the consistency and repeatability of the detection level. At the same time, we need to pay attention to the specificity and affinity of the antibody. Antibody specificity is mainly reflected in ensuring that the antibody only binds to the target antigen and reduces false positives caused by non-specific binding. It is not good if the antibody affinity is too high or too low. Too high may make it difficult to dissociate the antigen-antibody complex, affecting the results; too low will lead to unstable binding and weak signal. Antibodies with appropriate affinity constants should be selected. Another thing that needs to be considered is the expression of cell surface antigens (differences in expression levels, co-expression). For highly expressed antigens, low-concentration antibodies are selected, and for low-expressed antigens, high-concentration or high-affinity antibodies are selected to improve the detection sensitivity and avoid mutual interference in expression.

[0032] 2. Panel composition

[0033]

[0034] Note: * indicates detection of intracellular proteins.

[0035] Among them, the characteristics of each antibody are as follows:

[0036] CD5: A member of the scavenger receptor superfamily. CD5 is expressed on T cells, thymocytes, B cell subsets, chronic B lymphocytic leukemia (B cells), and peripheral blood dendritic cells.

[0037] CD7: It is a member of the immunoglobulin superfamily and is present in T cells, NK cells, thymocytes, hematopoietic progenitor cells and monocytes (to a weaker extent).

[0038] CD45: It is a tyrosine phosphatase expressed on the plasma membrane of all hematopoietic cells except red blood cells and platelets.

[0039] TRBC1: The T cell receptor (TCR) is a key molecule on the surface of T cells, responsible for recognizing antigenic peptide-MHC complexes on the surface of antigen-presenting cells (APCs) and initiating specific immune responses. TCRs are composed of two peptide chains and are divided into αβTCRs (approximately 95%) and γδTCRs (approximately 5%) based on chain type. The TRBC1 gene is one of the constant region genes of the β chain (TCRβ) of the γδTCR and is primarily involved in the development and functional regulation of γδT cells.

[0040] CD30: CD30 is highly expressed on Hodgkin cells, Reed-Sternberg cells, and activated (but not resting) T and B cells.

[0041] CD117: It is the receptor for stem cell factor or c-kit ligand. CD117 is expressed on multipotent hematopoietic progenitor cells (approximately 1-4% of bone marrow cells), mast cells, and acute myeloid leukemia (AML) cells.

[0042] CD56: Also known as NCAM (neural cell adhesion molecule), it is a single transmembrane glycoprotein. It is a member of the Ig superfamily. The 140 kD isoform is expressed on NK and NKT cells. CD56 is also expressed by certain large granular lymphocyte (LGL) leukemias, small cell lung cancer, neuronal-derived tumors, myelomas, and myeloid leukemias.

[0043] CD4: Also known as T4, expressed on most thymocytes.

[0044] CD2: is a 50kD type I transmembrane glycoprotein, also known as LFA-2, T11, and sheep erythrocyte receptor (SRBC-R). This member of the immunoglobulin superfamily is expressed on thymocytes, T lymphocytes, NK cells, and thymic B cell subsets.

[0045] CD3: It is a 20kD chain of the CD3 / T cell receptor (TCR) complex, composed of two CD3ε, one CD3γ, one CD3δ, one CD3ζ (CD247), and a T-Cell receptor (α / β or γ / δ) dimer. It is expressed on all mature T cells, NKT cells, and some thymocytes.

[0046] CD8: also known as T8 and LEU2, is expressed on most thymocytes and mainly on peripheral blood T cell subsets and NK cells.

[0047] Kappa: Each human immunoglobulin (Ig) antibody is composed of two identical heavy chains and two identical light chains. Kappa (κ) and lambda (λ) are two clonal molecules expressed on the membranes of mature B lymphocytes. Plasma cells often express them both intracellularly and on the cell membrane. The light chain (κ) is an important marker reflecting polyclonality.

[0048] Lambda: Each human immunoglobulin (Ig) molecule is composed of two identical heavy chains and two identical light chains. Kappa (κ) and lambda (λ) are two clonal molecules expressed on the membranes of mature B lymphocytes. Plasma cells often express them both intracellularly and on the cell membrane. The light chain (λ) is an important marker of polyclonality.

[0049] CD38: It is an ADP-ribosyl hydrolase that exhibits a variability in intensity on hematopoietic cells and certain non-hematopoietic tissues such as brain, muscle, and kidney.

[0050] CD10: It is a type II transmembrane protein. CD10 is expressed on B cell precursors, T cell precursors, and neutrophils.

[0051] CD19: A member of the immunoglobulin superfamily expressed on B cells (from Pro-B to Blastoid B cells, absent on plasma cells) and follicular dendritic cells.

[0052] CD20: Also known as B1 and BP35, it is a four-transmembrane protein expressed on mature B cells but not on normal plasma cells.

[0053] CD13: A 150-170 kb type I transmembrane glycoprotein also known as aminopeptidase N, APN, and GP150. This zinc metallopeptidase is expressed as a homodimer on the surface of granulocytes, myeloid progenitor cells, endothelial cells, epithelial cells, and some granulocyte lymphocytes, but not on platelets or red blood cells.

[0054] CD11b is a 165-170 kD type I transmembrane glycoprotein also known as αM integrin, Mac-1, CR3, and C3biR. CD11b non-covalently binds to integrin β2 (CD18) and is expressed on the surface of granulocytes, monocytes / macrophages, dendritic cells, natural killer cells (NK cells), and some T and B cells.

[0055] CD14: It is a 53-55kD glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that is often expressed at high levels on the surface of monocytes and macrophages, and at lower levels on the surface of granulocytes.

[0056] CD64: A 72kD single-chain type I glycoprotein, also known as FcγRI and FcRI. As a member of the immunoglobulin superfamily, CD64 is expressed on the surface of monocytes / macrophages, dendritic cells, and activated granulocytes.

[0057] CD33 is a 67kD type I transmembrane glycoprotein also known as sialic acid-binding immunoglobulin-like lectin 3 (Siglec-3), glycoprotein 67 (gp67), and p67. As a member of the sialic acid adhesion immunoglobulin superfamily, it is expressed on the surface of myeloid progenitor cells, monocytes, granulocytes, dendritic cells, and mast cells, but not on normal platelets, lymphocytes, red blood cells, or hematopoietic stem cells.

[0058] CD34, also known as gp105-120, is a type I monomeric sialomucin-like glycophosphoprotein with a molecular weight of approximately 105-120 kDa. It is selectively expressed in most hematopoietic stem / progenitor cells, bone marrow stromal cells, capillary endothelial cells, embryonic fibroblasts, and some neural tissues. It is a commonly used marker for identifying human hematopoietic stem / progenitor cells.

[0059] HLADR: A heterodimeric cell surface glycoprotein composed of a 36kD α (heavy) chain and a 27kD β (light) chain. It is expressed on the surface of B cells, activated T cells, monocytes / macrophages, dendritic cells, and other non-professional antigen-presenting cells (APCs).

[0060] The test results of each tube are analyzed as follows:

[0061] (1) T tube: It mainly depends on the differentiation and development rules of T / NK cell subsets and antibody combination to divide T lymphocytes differentiated from bone marrow hematopoietic stem cells into immature stages (T lymphocyte blast stage (CD7 bri +CD3-CD2-CD4-CD8-), thymic cortex stage (CD7 bri +CD3-CD2-CD4+CD8+), thymic medullary stage (CD7 bri +CD3+CD2+CD4+ / CD8+)) as well as mature stage (CD7+CD3+CD2+CD4+ / CD8+) and NK cell (CD3-CD56+) populations.

[0062] The use of other tumor expression auxiliary markers (CD30, CD117, TRBC1, CD5) combined with the antibody arrangement in the T tube can distinguish the difference in normal T cell and abnormal T cell antigen expression, judge T lymphocyte tumors, and improve the staging of T lymphoblastic lymphoma, the determination of mature T cell lymphoma subtypes, NK cell lymphoma and non-hematopoietic tumor auxiliary diagnosis. T lymphoblastic lymphoma often CD7 bri, decreased or heterogeneous expression of CD45, and CD2, CD4, CD8, and CD3 are used to differentiate naive T lymphocytes. Mature T cell lymphomas often rely on the expression and intensity of CD7, CD2, CD5, CD30, and CD117, in conjunction with TRBC1 monoclonal expression. Furthermore, CD3 and CD56 can differentiate NK cells, and combined with CD45 intensity, they facilitate initial screening for NK cell lymphomas. Decreased CD45 expression and increased CD56 expression are often seen in abnormal plasma cells and non-hematopoietic tumor cells.

[0063] (2) B tube: It mainly depends on the differentiation and development rules of B cell subsets and antibody combination to divide B lymphocytes differentiated from bone marrow hematopoietic stem cells into immature B cell group (CD19dim+CD20-CD38+), mature B lymphocyte group (CD19+CD20+CD38+ / -) and plasma cell group (CD19+ / -CD20-CD38 bri +). At the same time, CD10, CD56, and the cytoplasmic light chain binding antibody array in the B tube are combined to distinguish the differences in antigen expression between normal and abnormal B cells, determine B lymphocyte tumors, and distinguish immature B lymphocytes, mature B cell lymphomas, and plasma cell tumors.

[0064] In immature B lymphocytes (CD19dim+CD20-CD38+), the abnormal immature B lymphocyte stage (Pro / Common / Pre) can be inferred by combining CD10 expression and its intensity. In mature B lymphocytes (CD19+CD20+CD38+ / -), light chain restriction (cKappa or cLambda single positive) is an important indicator for judging clonality, and monoclonal proliferation usually indicates a tumor. CD5 is usually associated with chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL) and mantle cell lymphoma (MCL), and CD10 is more common in follicular lymphoma (FL) and Burkitt lymphoma. In addition, CD38 is often found in multiple myeloma (MM) or plasmacytoma. bri +, CD56+, but CD19 / CD20 are usually absent.

[0065] (3) M tube: Mainly relying on the differentiation and development rules of myeloid-derived cells and antibody combinations, myeloid cells differentiated from bone marrow hematopoietic stem cells are divided into primitive cells, granulocytes (neutrophils, eosinophils, basophils), monocytes, nucleated red blood cells and other myeloid cells (mast cells, etc.).

[0066] CD34, CD38, and HLA-DR are commonly markers of stem / progenitor cells and are crucial for the diagnosis of acute myeloid leukemia (AML). CD117, CD13, CD11b, CD33, CD14, and CD64 are key components of the myeloid lineage score in the EGIL (1995 edition) and WHO (2008 / 2016 edition) diagnostic criteria. High expression of CD14 and CD64 may indicate monocytic differentiation, while the combination of CD11b and CD13 may implicate granulocytic differentiation. Acute myeloid leukemia staging can be determined based on the expression and intensity of each antigen on the panel, combined with CD45 expression. Myelodysplastic neoplasms and chronic myeloid proliferative neoplasms can also be inferred by the granulocyte (CD13 / CD11b developmental pattern) and the proportion of myeloid blasts, combined with clinical history. Furthermore, for patients with anemia, mapping of CD45-negative nucleated red blood cells can prompt further testing for megaloblastic anemia.

[0067] Example 1

[0068] 1. Sample selection

[0069] 1.1 Samples collected using sodium heparin or EDTA anticoagulant tubes.

[0070] Exclusion criteria:

[0071] 1) Repeated sample enrollment.

[0072] 2) Samples that are visibly hemolyzed.

[0073] 3) The sample is visibly coagulated.

[0074] 1.2 Cell lines cultured in cell culture medium.

[0075] 2. Antibody preparation: Design the panel according to the T tube, B tube, and M tube channels and premix the antibodies in sequence. Premixed antibodies should be stored in a 4°C refrigerator away from light. Premixed reagents should be rewarmed to room temperature before reuse.

[0076] 3. Sample tube labeling: Number the sample tubes according to the sample number and experimental panel, such as: 1-T tube, 1-B tube, 1-M tube.

[0077] 4. Antibody and sample mixing: Add the premixed panel antibody into the test tube and mix with the premixed reagent.

[0078] 5. Antibody incubation: Mix at low speed for 3 seconds and incubate at room temperature in the dark for 15 minutes;

[0079] Red blood cell lysis: Add 1 ml of red blood cell lysis buffer to each sample tube, mix well, and incubate at room temperature in the dark for 10 minutes (no more than 15 minutes is recommended).

[0080] 6. Wash the sample: Centrifuge at room temperature at 1500 rpm for 5 minutes.

[0081] 7. If the sample does not require intracellular detection, repeat the above steps twice and prepare for flow cytometer detection. If intracellular target detection is required (ckappa, clambda), continue with the following fixation and membrane permeabilization operations.

[0082] 8. Cell membrane disruption: Discard the supernatant, add 100 μl of membrane disruption reagent, and incubate in BD Intrasure at room temperature in the dark. TM Reagent A for 6 min, centrifugation at room temperature at 1500 rpm for 5 min;

[0083] 9. Intracellular staining: Discard the supernatant and add 50ul of BD Intrasure TM Reagent B and intracellular antibody were mixed at low speed for 3 seconds and incubated at room temperature in the dark for 15 minutes.

[0084] 10. Wash cells: Centrifuge at room temperature at 1500 rpm for 5 minutes.

[0085] 11. Resuspend cells: Discard the supernatant and resuspend in 0.2 ml of PBS. Mix at low speed for three seconds. Protect from light while waiting for analysis. If analysis is not possible immediately, store in a refrigerator at 2-8°C, away from light, and analyze within 24 hours.

[0086] 12. Obtain data on the computer.

[0087] 13. Complete the result interpretation.

[0088] Figure 2-4 is a two-dimensional dot plot of flow cytometry, where:

[0089]

[0090] Obviously, the above embodiments are merely examples for clarity of explanation and are not intended to limit the implementation methods. Those skilled in the art will readily appreciate that other variations or modifications based on the above descriptions are possible. It is not necessary and impossible to enumerate all implementation methods here. Obvious variations or modifications arising therefrom remain within the scope of protection of the present invention.

Claims

1. A flow cytometry panel for the initial screening of hematologic tumors, characterized by: It includes a T tube, a B tube, and an M tube; the T tube includes 10 channels composed of the following antibodies: CD5, CD7, CD45, TRBC1, CD30 / 117, CD56, CD4, CD2, CD3, and CD8; The B tube includes 9 channels composed of the following antibodies: CD5, CD56, CD45, cKappa, cLambda, CD38, CD10, CD19, and CD20; The M tube includes 10 channels composed of the following antibodies: CD13, CD11b, CD45, CD14, CD64, CD33, CD117, CD34, HLA DR and CD38.

2. The flow detection panel according to claim 1, characterized in that: The hematological tumors include leukemia, lymphoma, multiple myeloma, paroxysmal nocturnal hemoglobinuria, chronic bone proliferative tumors, myeloproliferative neoplasms and megaloblastic anemia.

3. Use of the flow cytometry panel according to claim 1 in preparing a device for primary screening of hematological tumors.

4. Use of the flow cytometry panel according to claim 1 in the preparation of a primary screening reagent for hematological tumors.

5. A flow cytometry kit for the primary screening of hematological tumors, characterized by: The kit contains antibodies labeled with different fluorescein, which are divided into three groups. The first group consists of the following antibodies: CD5, CD7, CD45, TRBC1, CD30 / 117, CD56, CD4, CD2, CD3 and CD8; The second panel consisted of the following antibodies: CD5, CD56, CD45, cKappa, cLambda, CD38, CD10, CD19, and CD20; The third panel consisted of the following antibodies: CD13, CD11b, CD45, CD14, CD64, CD33, CD117, CD34, HLA DR, and CD38; Each antibody in each set is packaged separately.

6. A detection reagent for the initial screening of hematological tumors, characterized by: The detection reagents include antibodies labeled with different fluorescein, and the antibodies are divided into three groups. The first group consists of the following antibodies: CD5, CD7, CD45, TRBC1, CD30 / 117, CD56, CD4, CD2, CD3 and CD8; The second panel consisted of the following antibodies: CD5, CD56, CD45, cKappa, cLambda, CD38, CD10, CD19, and CD20; The third panel consisted of the following antibodies: CD13, CD11b, CD45, CD14, CD64, CD33, CD117, CD34, HLA DR, and CD38; Each antibody in each set is packaged separately.

7. A system for primary screening of hematological tumors, characterized by: It includes detection part and analysis part; The detection part includes the detection reagent according to claim 6; The analyzing part is used to analyze the detection results of the detecting part.

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