Rapid screening reagent combination and application for mature lymphocytic tumors

Abstract

The present invention discloses a rapid screening reagent combination and application of mature lymphocyte tumors. The rapid screening reagent combination of the present invention is an antibody composition, which includes: anti-cytoplasmic κ antibody, anti-CD4 antibody, anti-cytoplasmic λ antibody, anti-CD8 antibody, anti-CD3 antibody, anti-CD19 antibody, anti-CD20 antibody, anti-CD56 antibody, anti-CD5 antibody, anti-CD10 antibody, anti-CD38 antibody, anti-CD57 antibody, and anti-CD45 antibody. The rapid screening reagent combination of mature lymphocyte tumors of the present invention can quickly and accurately realize the immunotyping of lymphocyte tumors.

Description

Technical Field

[0001] The invention relates to a rapid screening reagent combination for mature lymphocyte tumors and application thereof. Background Art

[0002] Normal peripheral blood mature lymphocytes include T cells, B cells and NK cells. Clinical diseases characterized by proliferation of these three types of mature lymphocytes in peripheral blood mainly include chronic lymphocytic leukemia (CLL), large granular T-lymphocytic leukemia (T-LGLL) and NK cell chronic lymphoproliferative disease (NK-CLPD).

[0003] The typical immunophenotype of CLL is usually CD19+, CD5+, CD23+, CD200+, CD10-, FMC7-, CD43+ / -, CCND1-, with weak expression (dim) of surface immunoglobulin (sIg), CD20 and CD79b.

[0004] Normal mature circulating B cells express CD19, CD20, CD79b, CD22, FMC7 and surface immunoglobulin (sIg). The ratio of light chain kappa (sIgκ) to lambda (sIgλ) of sIg is between 1:3 and 3:1.

[0005] Flow cytometry confirmed the clonality of B cells by restricted expression of κ or λ light chains on the B cell surface (κ:λ > 3:1 or < 0.3:1) or the absence of sIg expression by > 25% of B cells.

[0006] NK-CLPD is a type of large granular lymphocytic leukemia. Large granular lymphocytic proliferative disease refers to a group of diseases in which peripheral blood LGLs continue to increase without a clear cause. Large granular lymphocytes (LGL) are a subset of lymphocytes, whose cell bodies are larger than normal lymphocytes and have rich cytoplasm. LGLs account for 10% to 15% of normal peripheral blood mononuclear cells, including two cell populations, CD3+ and CD3-: CD3+ (T cells), namely T-LGL, CD3+-LGL, which are activated cytotoxic lymphocytes; CD3- (NK cells), namely NK-LGL, CD3-LGL.

[0007] REAL classification divides LGLL into T-LGLL and NK-LGLL. T-LGLL is CD3+ clonal proliferation, with the main phenotype of CD3+, TCRαβ+, CD4-, CD8+, and T cell receptor rearrangement is monoclonal. NK-LGLL is CD3- NK cell clonal proliferation, cytogenetics or KIR examination can prove its clonality, but the uniform expression of CD56 / CD57 is suggestive of abnormal NK.

[0008] It can be seen that the immune phenotypes of mature lymphocytic tumors are diverse, and therefore the immunophenotyping of mature lymphocytic tumors remains a difficult problem faced in clinical practice. Summary of the invention

[0009] In order to overcome the above disadvantages, the purpose of the present invention is to provide a rapid screening reagent combination and application for mature lymphocyte tumors.

[0010] The rapid screening reagent combination for mature lymphocyte tumors of the present invention comprises a first antibody composition and a second antibody composition, wherein the first antibody composition comprises: anti-cytoplasmic κ antibody, anti-CD4 antibody, anti-cytoplasmic λ antibody, anti-CD8 antibody, anti-CD3 antibody, anti-CD19 antibody, anti-CD20 antibody, anti-CD56 antibody, anti-CD5 antibody, anti-CD10 antibody, anti-CD38 antibody, anti-CD57 antibody, and anti-CD45 antibody; and the second antibody composition comprises anti-CD4 antibody, anti-CD8 antibody, anti-CD3 antibody, anti-TRBC1 antibody, and anti-CD45 antibody.

[0011] Furthermore, the antibody composition is a fluorescein-labeled antibody, and the individual antibodies of the first antibody composition are paired with the following labeled fluorescence in sequence: KAPPA-FITC, CD4-FITC, CD8-PE, CD3-APC750, LAMBDA-FITC, CD38-ECD, CD19-APC700, CD20-PC5, CD56-PC5, CD5-PC7, CD10-APC, CD57-PB, CD45-Kro; the individual antibodies of the second antibody composition are paired with the following labeled fluorescein in sequence: CD4-APC750, CD8-PE, CD3-ECD, TRBC1-FITC and CD45-KRO.

[0012] Furthermore, the antibody composition is a fluorescein-labeled antibody, and the ratio of the added amount of each fluorescein-labeled antibody is: KAPPA-FITC 1, CD4-FITC 5, CD8-PE 5, CD3-APC750 3, LAMBDA-FITC 1, CD38-ECD 3, CD19-APC700 3, CD20-PC5 5, CD56-PC5 5, CD5-PC7 5, CD10-APC 3, CD57-PB 5, CD45-Kro 3.

[0013] Further, the added amount of each of the first antibody compositions is: KAPPA-FITC 1, CD4-FITC 5, CD8-PE 5, CD3-APC750 3, LAMBDA-FITC 1, CD38-ECD 3, CD19-APC700 3, CD20-PC5 5, CD56-PC5 5, CD5-PC7 5, CD10-APC 3, CD57-PB 5, CD45-Kro 3;

[0014] The added amounts of each of the first antibody compositions are: 5 for CD4-APC750, 5 for CD8-PE, 5 for CD3-ECD, 5 for TRBC1-FITC, and 5 for CD45-KRO.

[0015] The present invention also provides a kit, which comprises the aforementioned antibody composition.

[0016] The present invention also provides the use of the reagent composition in preparing flow cytometry samples for rapid screening of mature lymphocyte tumors.

[0017] Furthermore, the preparation process of the flow cytometer sample for rapid screening of mature lymphocyte tumors includes the following steps:

[0018] ① Collection and pretreatment of samples to be tested: collect peripheral blood, lyse red blood cells, centrifuge, and discard the supernatant;

[0019] ② Preparation of the first antibody composition reagent: dissolving each antibody of the first antibody composition in a buffer solution for later use;

[0020] Preparation of the second antibody composition reagent: dissolving each antibody of the second antibody composition in a buffer solution for later use;

[0021] ③Sample processing:

[0022] Take an appropriate amount of the sample prepared in step ① and add the first antibody composition reagent prepared in step ②, incubate in the dark; add an appropriate amount of buffer, centrifuge, discard the supernatant, add an appropriate amount of buffer, centrifuge, discard the supernatant, add an appropriate amount of buffer to suspend, and then detect on the machine.

[0023] Furthermore, the gates are set according to the following steps when the flow cytometer is used for detection:

[0024] Step 1: Use CD45 / SS INT to set the gate and circle the lymphocyte B gate;

[0025] Step 2: Use CD3 and CD19 to set gates in Gate B to circle out T cells, mature B cells, and NK cells;

[0026] Step 3: Use KAPPA / CD4 / LAMBDA / CD8 to set the gate and circle out mature T cells. If the ratio between CD4+ and CD8+ is greater than 5, add TRBC1;

[0027] Step 4: Use CD20+CD56 / CD5 in the CD19+ gate obtained in step 2 to circle the K gate. The cells in the K gate are CD5+ mature B lymphocytes.

[0028] Step 5: Use KAPPA / CD4 / LAMBDA / CD8 to set the gate in the K gate, and determine the type of monoclonal B lymphocytes based on the ratio between KAPPA / CD4 and LAMBDA / CD8:

[0029] If LAMBDA / KAPPA>3, it indicates that it is a LAMBDA type monoclonal B lymphocyte; if KAPPA / LAMBDA>3, it indicates that the B lymphocytes in the sample are KAPPA type monoclonal B lymphocytes.

[0030] Furthermore, in step 2, the immunophenotype of the T cells is CD3+, the cell phenotype of the mature B cells is CD19+, and the cell phenotype of the NK cells is CD3- and CD19-.

[0031] Furthermore, the step of adding TRBC1 is: after the sample is subjected to red lysis treatment, the second antibody composition is added, followed by incubation and washing before processing on the machine. If the antibody positive ratio TRBC1 is <15% or >85%, it is abnormal and may be monoclonal T-cell lymphoma; if 15% <antibody positive ratio TRBC1 <85%, it is normal.

[0032] Furthermore, the antibody positive ratio TRBC1 is obtained according to the following steps:

[0033] Step 2-1: First use the CD3 / CD19 gate to find CD3+ positive cells in gate B;

[0034] Step 2-2: Display KAPPA / CD4+TRBC1 / LAMBDA / CD8 in the CD3+ gate,

[0035] Step 2-3: Find CD8+ and CD4+ positive cells;

[0036] Step 2-4: If CD8+ / CD4+>5, select CD8+ cells and use CD8+ cells to set the gate.

[0037] KAPPA / CD4+TRBC1 was displayed in the gate, and the proportion of the KAPPA / CD4+TRBC1-positive fraction was subsequently calculated;

[0038] If CD4+ / CD8+>5, CD4+ cells were selected, and the gate was set using CD4+ cells. KAPPA / CD8+TRBC1 was displayed in the gate, and then the proportion of the KAPPA / CD8+TRBC1 positive part was calculated. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] FIG1 (a) is a sample flow cytometry gating analysis result of Example 1 of the present invention;

[0040] FIG1( b ) is a sample flow cytometry gating analysis result of Example 1 of the present invention;

[0041] FIG2 (a) is a sample flow cytometry gating analysis result of Example 2 of the present invention;

[0042] FIG2( b ) is a sample flow cytometry gating analysis result of Example 2 of the present invention;

[0043] FIG3 (a) is a sample flow cytometry gating analysis result of Example 3 of the present invention;

[0044] FIG3 (b) is a sample flow cytometry gating analysis result of Example 3 of the present invention;

[0045] FIG4 (a) is a sample flow cytometry gating analysis result of Example 4 of the present invention;

[0046] FIG4( b ) is a sample flow cytometry gating analysis result of Example 4 of the present invention;

[0047] FIG5 (a) is a result obtained by setting gates for sample flow cytometry analysis in Example 5 of the present invention;

[0048] FIG5( b ) is the result of gating analysis of sample flow cytometry in Example 5 of the present invention;

[0049] FIG5 (c) is the flow cytometry gating analysis result of adding TRBC1 to the sample in Example 5 of the present invention;

[0050] FIG6 (a) is a result obtained by setting gates for sample flow cytometry analysis in Example 6 of the present invention;

[0051] FIG6 (b) is the result of gating analysis of sample flow cytometry in Example 6 of the present invention;

[0052] FIG6 (c) is the flow cytometry gating analysis result of adding TRBC1 to the sample according to Example 6 of the present invention. DETAILED DESCRIPTION

[0053] The preferred embodiments of the present invention are described in detail below in conjunction with the accompanying drawings so that the advantages and features of the present invention can be more easily understood by those skilled in the art, thereby making a clearer and more definite definition of the protection scope of the present invention.

[0054] In some embodiments, the sample analysis using the reagent composition of the present invention is performed according to the following steps:

[0055] 1. Sample collection and pretreatment:

[0056] Take 200 μl of the patient's peripheral blood, add 2 mL of ammonium chloride hemolytic agent, vortex and mix, let stand for 15 minutes; add 5 ml PBS and centrifuge for 5 minutes / 1500 rpm to wash twice, and discard the supernatant;

[0057] 2. Preparation of antibody composition reagents:

[0058] Each antibody composition is a fluorescein-labeled antibody, and each antibody of the first antibody composition is paired with the following labeled fluorescence in sequence: KAPPA-FITC, CD4-FITC, CD8-PE, CD3-APC750, LAMBDA-FITC, CD38-ECD, CD19-APC700, CD20-PC5, CD56-PC5, CD5-PC7, CD10-APC, CD57-PB, and CD45-Kro.

[0059] Each first antibody composition (anti-cytoplasmic κ antibody, anti-CD4 antibody, anti-cytoplasmic λ antibody, anti-CD8 antibody, anti-CD3 antibody, anti-CD19 antibody, anti-CD20 antibody, anti-CD56 antibody, anti-CD5 antibody, anti-CD10 antibody, anti-CD38 antibody, anti-CD57 antibody, anti-CD45 antibody) was dissolved in a buffer solution.

[0060] Each antibody of the second antibody composition is conjugated with the following fluorescent markers in sequence: CD4-APC750, CD8-PE, CD3-ECD, TRBC1-FITC and CD45-KRO.

[0061] In some embodiments, the ratio of each first antibody solution in the first antibody composition is: KAPPA-FITC 1, CD4-FITC 5, CD8-PE 5, CD3-APC750 3, LAMBDA-FITC 1, CD38-ECD 3, CD19-APC700 3, CD20-PC5 5, CD56-PC5 5, CD5-PC7 5, CD10-APC 3, CD57-PB 5, CD45-Kro 3. The ratio of each antibody is the volume ratio after each fluorescent-labeled antibody is dissolved in a buffer solution.

[0062] The ratio of each second antibody solution in the second antibody composition is: CD4-APC750 5, CD8-PE 5, CD3-ECD 5, TRBC1-FITC 5, CD45-KRO 5.

[0063] 3. Sample processing: Add the antibody composition reagent prepared in step 2 to the washed cells (50-100 μl) prepared in step ①, mix well, and incubate at room temperature for 30 min; centrifuge and wash with 2 ml PBS for 5 min / 1500 rpm, discard the supernatant, add 500 μl PBS to the cell pellet for suspension, and detect on the machine.

[0064] 4. Data Analysis

[0065] Different samples were processed according to the above method and analyzed by flow cytometry. The specific flow cytometry analysis results are shown in Example 1-2.

[0066] When performing flow cytometry analysis, gate the samples according to the following steps:

[0067] Step 1: Use CD45 / SS INT to set the gate and circle the lymphocyte B gate;

[0068] Step 2: Use CD3 and CD19 to set gates in Gate B to circle out T cells, mature B cells, and NK cells, wherein the immunophenotype of the T cells is CD3+, the cell phenotype of the mature B cells is CD19+, and the cell phenotype of the NK cells is CD3- and CD19-;

[0069] Step 3: Use KAPPA / CD4 / LAMBDA / CD8 to set the gate and circle out mature T cells. If the ratio between CD4+ and CD8+ is greater than 5, add TRBC1;

[0070] Step 4: Use CD20+CD56 / CD5 in the CD19+ gate obtained in step 2 to circle the K gate. The cells in the K gate are CD5+ mature B lymphocytes.

[0071] Step 5: Use KAPPA / CD4 / LAMBDA / CD8 to set the gate in the K gate, and determine the type of monoclonal B lymphocytes based on the ratio between KAPPA / CD4 and LAMBDA / CD8:

[0072] If LAMBDA / KAPPA>3, it indicates that it is a LAMBDA type monoclonal B lymphocyte; if KAPPA / LAMBDA>3, it indicates that the B lymphocytes in the sample are KAPPA type monoclonal B lymphocytes.

[0073] In some embodiments, the steps of adding TRBC1 are as follows: Take 100 μL of blood sample, lyse red blood cells, add 5 μL each of CD4-APC750, CD8-PE, CD3-ECD, TRBC1-FITC, and CD45-KRO, then incubate for 15 min, wash with PBS, and load onto the machine.

[0074] If the positive antibody ratio of TRBC1 < 15% or > 85%, it is abnormal and may be monoclonal T-cell lymphoma; if 15% < TRBC1 < 85%, it is normal.

[0075] In some embodiments, the clone numbers of the respective antibodies of the present invention are as follows: the clone number of the anti-cytoplasmic κ antibody is GAHK, the clone number of the anti-CD4 antibody is 13B8.2, the clone number of the anti-cytoplasmic λ antibody is GAHL, the clone number of the anti-CD8 antibody is B9.11, the clone number of the anti-CD3 antibody is UCHT1, the clone number of the anti-CD19 antibody is J3.119, the clone number of the anti-CD20 antibody is B9E9, the clone number of the anti-CD56 antibody is N901, the clone number of the anti-CD5 antibody is BL1a, the clone number of the anti-CD10 antibody is ALB1, the clone number of the anti-CD38 antibody is LS198-4-3, the clone number of the anti-CD57 antibody is NC1, and the clone number of the anti-CD45 antibody is J33;

[0076] The second antibody composition includes the anti-CD4 antibody with a clone number of 13B8.2, the anti-CD8 antibody with a clone number of B9.11, the anti-CD3 antibody with a clone number of UCHT1, and the anti-CD45 antibody with a clone number of J33.

[0077] Example 1:

[0078] Referring to FIGS. 1(a) and 1(b), when the sample in this example is subjected to flow cytometry analysis, the proportion of lymphocytes in gate B circled by using CD45 / SS INT gating is 89.97%;

[0079] Subsequently, using CD3 and CD19 for gating, the proportions of T cells, mature B cells, and NK cells circled are 1.49%, 98.13%, and 0.15% respectively;

[0080] Subsequently, using KAPPA / CD4 / LAMBDA / CD8 for gating to circle mature T cells;

[0081] Subsequently, in mature B cells (CD19+ gate), using CD20+CD56 / CD5 for gating to circle gate K, and the cells in gate K are CD5+ mature B lymphocytes;

[0082] Using KAPPA / CD4 / LAMBDA / CD8 as the gate in the K gate, we can obtain the ratio between KAPPA / CD4 and LAMBDA / CD8 as LAMBDA / KAPPA=39.44% / 1.92%=20.5>3. Therefore, the immunophenotype of this sample is CD5+, CD19+, CD23+, FMC-7-, CD103-, CD200+, CD79B-, CD10-, CD20+, CD22-, CD38-, LAMBDA / KAPPA>3, indicating a LAMBDA type CD5+ monoclonal B lymphocytoma.

[0083] Embodiment 2:

[0084] As shown in FIG. 2 (a) and FIG. 2 (b), when the sample in this embodiment is subjected to flow cytometric analysis, the proportion of lymphocytes in gate B circled by CD45 / SS INT is 87.22%;

[0085] Subsequently, CD3 and CD19 were used to set gates, and the proportions of T cells, mature B cells, and NK cells were 7.30%, 91.30%, and 1.35%, respectively;

[0086] Subsequently, KAPPA / CD4 / LAMBDA / CD8 were used to set gates to circle out mature T cells;

[0087] Then, among the mature B cells (CD19+ gate), use CD20+CD56 / CD5 to circle the K gate, and the cells in the K gate are CD5+ mature B lymphocytes;

[0088] Using KAPPA / CD4 / LAMBDA / CD8 as the gate in the K gate, we can obtain the ratio between KAPPA / CD4 and LAMBDA / CD8 as KAPPA / LAMBDA=21.74% / 0.08%=272>3. Therefore, the immunophenotype of this sample is: CD5+, CD19+, CD23+, FMC-7-, CD103-, CD200+, CD79B-, CD10-, CD20+, CD22-, CD38-, KAPPA / LAMBDA>3, indicating a KAPPA type CD5+ monoclonal B lymphocytoma.

[0089] Embodiment 3:

[0090] As shown in FIG. 3 (a) and FIG. 3 (b), when the sample in this embodiment is subjected to flow cytometric analysis, the proportion of lymphocytes in gate B circled by CD45 / SS INT is 53.29%;

[0091] Subsequently, CD3 and CD19 were used to set gates, and the proportions of T cells, mature B cells, and NK cells were 3.08%, 95.88%, and 1.04%, respectively;

[0092] Subsequently, KAPPA / CD4 / LAMBDA / CD8 were used to set gates to circle out mature T cells;

[0093] Then, among the mature B cells (CD19+ gate), use CD20+CD56 / CD5 to circle the K gate, and the cells in the K gate are CD5+ mature B lymphocytes;

[0094] KAPPA / CD4 / LAMBDA / CD8 were used to set the gate in the K gate, and the ratio between KAPPA / CD4 and LAMBDA / CD8 was KAPPA / LAMBDA=0.97% / 0.92%=1<3. Therefore, the immunophenotype of this sample was CD5+, CD19+, CD23+, FMC-7-, CD103-, CD200++, CD79B-, CD10-, CD20+, CD22-, CD38-, KAPPA / LAMBDA<3, which was a KAPPA-negative and LAMBDA-negative clonal B lymphocytic neoplasm.

[0095] Embodiment 4:

[0096] As shown in FIG. 4 (a) and FIG. 4 (b), when the sample in this embodiment is subjected to flow cytometric analysis, the proportion of lymphocytes in gate B circled by CD45 / SS INT is firstly set to 10.85%;

[0097] Subsequently, CD3 and CD19 were used to set gates, and the proportions of T cells, mature B cells, and NK cells were 78.54%, 13.16%, and 3.50%, respectively;

[0098] Subsequently, KAPPA / CD4 / LAMBDA / CD8 were used to set gates to circle out mature T cells;

[0099] Then, among the mature B cells (CD19+ gate), use CD20+CD56 / CD5 to circle the K gate, and the cells in the K gate are CD5+ mature B lymphocytes;

[0100] The immunophenotype of this sample is: CD5+, CD19+, CD23+-, FMC-7-, CD103-, CD200+-, CD79B+-, CD10-, CD20+, CD22+-, CD38-. No abnormalities were found in this sample.

[0101] Embodiment 5:

[0102] Referring to Figures 5(a) and 5(b), after flow cytometry analysis of the samples, it was found that CD8 accounted for 82.05% and CD4 accounted for 8.96%, so CD8 / CD4>5. Therefore, it can be preliminarily judged that the result is abnormal. At this time, TRBC1 needs to be added.

[0103] See Figure 5 (c), which shows the flow cytometry analysis process and results of TRBC1:

[0104] First, use the CD3 / CD19 gate, that is, find CD3+ positive cells in gate B, accounting for 71.29%;

[0105] Then, KAPPA / CD4+TRBC1 / LAMBDA / CD8 were displayed in the CD3+ gate to find CD8+ and CD4+ positive cells;

[0106] Since CD8+ / CD4+=83.77% / 7.78%=10.77>5, CD8+ cells were selected and used to set the gate.

[0107] KAPPA / CD4+TRBC1 was displayed in the gate, and then the proportion of KAPPA / CD4+TRBC1 positive fraction was calculated.

[0108] The antibody positive ratio TRBC1 in this example is 21.06%, indicating that the sample is normal.

[0109] Embodiment 6:

[0110] Referring to Figures 6 (a) and 6 (b), after flow cytometry analysis of the samples, it was found that CD8 accounted for 79.84% and CD4 accounted for 13.65%, so CD8 / CD4=5.85>5. Therefore, it can be preliminarily judged that the result is abnormal. At this time, TRBC1 needs to be added.

[0111] See Figure 6 (c), which shows the flow cytometry analysis process and results of TRBC1:

[0112] First, use the CD3 / CD19 gate, that is, find CD3+ positive cells in gate B, accounting for 82.60%;

[0113] Then, KAPPA / CD4+TRBC1 / LAMBDA / CD8 were displayed in the CD3+ gate to find CD8+ and CD4+ positive cells;

[0114] Since CD8+ / CD4+=81.34% / 9.52%=28.54>5, CD8+ cells were selected and used to set the gate.

[0115] KAPPA / CD4+TRBC1 was displayed in the gate, and then the proportion of KAPPA / CD4+TRBC1 positive fraction was calculated.

[0116] The antibody positive rate TRBC1 in this example is 40.42%, which is preliminarily determined to be normal cells.

[0117] The above implementation modes are only for illustrating the technical concept and features of the present invention, and their purpose is to enable people familiar with this technology to understand the content of the present invention and implement it, and they cannot be used to limit the protection scope of the present invention. Any equivalent changes or modifications made according to the spirit of the present invention should be included in the protection scope of the present invention.

Claims

1. A rapid screening reagent combination for mature lymphocyte tumors, characterized in that: comprising a first antibody composition and a second antibody composition, The first antibody composition comprises: anti-cytoplasmic κ antibody, anti-CD4 antibody, anti-cytoplasmic λ antibody, anti-CD8 antibody, anti-CD3 antibody, anti-CD19 antibody, anti-CD20 antibody, anti-CD56 antibody, anti-CD5 antibody, anti-CD10 antibody, anti-CD38 antibody, anti-CD57 antibody, and anti-CD45 antibody; The second antibody composition comprises anti-CD4 antibody, anti-CD8 antibody, anti-CD3 antibody, anti-TRBC1 antibody and anti-CD45 antibody.

2. The rapid screening reagent combination for mature lymphocytic tumors according to claim 1, characterized in that: The antibody composition is a fluorescein-labeled antibody, and each antibody of the first antibody composition is paired with the following labeled fluorescence in sequence: KAPPA-FITC, CD4-FITC, CD8-PE, CD3-APC750, LAMBDA-FITC, CD38-ECD, CD19-APC700, CD20-PC5, CD56-PC5, CD5-PC7, CD10-APC, CD57-PB, CD45-Kro; Each antibody of the second antibody composition is sequentially paired with the following fluorescent markers: CD4-APC750, CD8-PE, CD3-ECD, TRBC1-FITC and CD45-KRO.

3. The rapid screening reagent combination for mature lymphocytic tumors according to claim 1, characterized in that: The ratio of the added amount of each of the first antibody compositions is: KAPPA-FITC 1, CD4-FITC 5, CD8-PE 5, CD3-APC750 3, LAMBDA-FITC 1, CD38-ECD 3, CD19-APC700 3, CD20-PC5 5, CD56-PC5 5, CD5-PC7 5, CD10-APC 3, CD57-PB 5, CD45-Kro3; The ratio of the added amount of each of the second antibody compositions is: CD4-APC750 5, CD8-PE 5, CD3-ECD 5, TRBC1-FITC 5, CD45-KRO 5.

4. A kit, characterized in that: The kit comprises the first antibody composition and the second antibody composition according to any one of claims 1 to 3.

5. Use of the rapid screening reagent combination for mature lymphocyte tumors according to any one of claims 1 to 3 in preparing flow cytometric samples for rapid screening of mature lymphocyte tumors.

6. The use according to claim 5, characterized in that: The preparation process of the flow cytometer sample for rapid screening of mature lymphocyte tumors comprises the following steps: ① Collection and pretreatment of samples to be tested: collect peripheral blood, lyse red blood cells, centrifuge, and discard the supernatant; ② Preparation of the first antibody composition reagent: dissolving each antibody of the first antibody composition in a buffer solution for later use; Preparation of the second antibody composition reagent: dissolving each antibody of the second antibody composition in a buffer solution for later use; ③Sample processing: Take an appropriate amount of the sample prepared in step ① and add the first antibody composition reagent prepared in step ②, incubate in the dark; add an appropriate amount of buffer, centrifuge, discard the supernatant, add an appropriate amount of buffer, centrifuge, discard the supernatant, add an appropriate amount of buffer to suspend, and then detect on the machine.

7. The use according to claim 6, characterized in that: When performing flow cytometry testing, set the gate according to the following steps: Step 1: Use CD45 / SS INT to set the gate and circle the lymphocyte B gate; Step 2: Use CD3 and CD19 to set gates in Gate B to circle out T cells, mature B cells, and NK cells; Step 3: Use KAPPA / CD4 / LAMBDA / CD8 to set the gate and circle out mature T cells. If the CD8 / CD4 ratio is greater than 5, add TRBC1; Step 4: Use CD20+CD56 / CD5 in the CD19+ gate obtained in step 2 to circle the K gate. The cells in the K gate are CD5+ mature B lymphocytes. Step 5: Use KAPPA / CD4 / LAMBDA / CD8 to set the gate in the K gate, and determine the type of monoclonal B lymphocytes based on the ratio between KAPPA / CD4 and LAMBDA / CD8: If LAMBDA / KAPPA>3, it indicates that it is a LAMBDA type monoclonal B lymphocyte; if KAPPA / LAMBDA>3, it indicates that the B lymphocytes in the sample are KAPPA type monoclonal B lymphocytes.

8. The use according to claim 7, characterized in that: In step 2, the immunophenotype of the T cells is CD3+, the cell phenotype of the mature B cells is CD19+, and the cell phenotype of the NK cells is CD3- and CD19-.

9. The use according to claim 8, characterized in that: The step of adding TRBC1 is: after the sample is subjected to red lysis treatment, the second antibody composition is added, followed by incubation and washing and then processing on the machine. If the antibody positive ratio TRBC1 is <15% or the antibody positive ratio TRBC1>85%, it is abnormal and may be monoclonal T-cell lymphoma; if 15%<antibody positive ratio TRBC1<85%, it is normal.

10. The use according to claim 9, characterized in that: The antibody positive ratio TRBC1 is obtained according to the following steps: Step 2-1: First use the CD3 / CD19 gate to find CD3+ positive cells in gate B; Step 2-2: Display KAPPA / CD4+TRBC1 / LAMBDA / CD8 in the CD3+ gate, Step 2-3: Find CD8+ and CD4+ positive cells; Step 2-4: If CD8+ / CD4+>5, select CD8+ cells and use CD8+ cells to set the gate. KAPPA / CD4+TRBC1 was displayed in the gate, and then the proportion of KAPPA / CD4+TRBC1 positive fraction was calculated. If CD4+ / CD8+>5, CD4+ cells were selected, and the gate was set using CD4+ cells. KAPPA / CD8+TRBC1 was displayed in the gate, and then the proportion of the KAPPA / CD8+TRBC1 positive part was calculated.

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