Reagent kit and method for detecting expression quantities of antigens of acute myelogenous leukemia granulocytes
A technology of acute myeloid and detection method, applied in acute myeloid leukemia granulocyte antigen expression detection kit, acute myeloid leukemia granulocyte antigen expression detection field, can solve few problems such as quantitative analysis of antibody expression, achieve Accurate and quantitative detection of the effect of the analysis
Inactive Publication Date: 2017-01-04
北京海思特医学检验实验室有限公司
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Flow cytometry can detect the forward scatter (Forward Scatter, FSC), side scatter (SSC) and the intensity of multiple fluorescent channels of the sample cells at the same time. The antibody expression of each group of cells is known, but the traditional flow cytometry is mainly to measure the proportion of cells expressing the target antigen, and to perform qualitative analysis on the expression of the target antigen, that is, to judge whether the antibody expression is negative, weakly positive or strong Positive, but rarely quantitative analysis of antibody expression
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[0047] As a preferred embodiment, the specific operation process is:
[0048] In step S1, the peripheral blood of the AML patient is taken as a test sample, the white blood cell count is performed on the test sample, and the cell concentration is adjusted to 3×10 7 / mL;
[0049] In step S2, take a dedicated flow tube, add 10 μL of CD45-Percp, 20 μL of CD15FITC, and 10 μL of CD33PE into the flow tube; add 70 μL of the test sample into the flow tube, and vortex at low speed to mix;
[0050] In step S3, incubate the mixed test specimen at room temperature in the dark for 20 minutes; add 1 mL of erythrocyte lysate solution to the flow tube, vortex at low speed to mix, and lyse the red blood cells at room temperature in the dark for 10 minutes; After the end, discard the supernatant, then add 1mL PBS to the tube, vortex at low speed to mix, and centrifuge at 1500r / min for 5min; Within 24 hours, use the flow cytometer to test on the machine; during the test, adjust the voltage of ...
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The invention provides a reagent kit and a method for detecting the expression quantities of antigens of acute myelogenous leukemia granulocytes. Reagents of the reagent kit comprise percp-labeled CD45-resistant antibodies, FITC-labeled CD15-resistant antibodies and PE-labeled CD33-resistant antibodies. The reagent kit is applied to the method, and the expression quantities of the CD33 antigens of the granulocytes (non-stem/progenitor cells) in peripheral blood and bone marrow of AML (acute myelogenous leukemia) patients can be detected by the aid of flow cytometry. Compared with the traditional flow cytometry mainly for measuring target expression antigen cell proportions and qualitatively analyzing the target expression antigen cell proportions, the reagent kit and the method have the advantages that the expression of the CD33 antigens of the myelogenous leukemia granulocytes can be quickly and accurately quantitatively detected and analyzed by the aid of the reagent kit and the method, detection results can be used for evaluating curative effects of AML monoclonal antibody therapy, targeted therapy for AML by the aid of CD33 monoclonal antibody medicines can be effectively guided, and extremely important effects can be realized in related medical detection fields.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for detecting the expression level of granulocyte antigens in acute myeloid leukemia (AML). The invention also relates to a method for detecting the expression level of the acute myeloid leukemia granulocyte antigen. Background technique [0002] AML is a clonal malignant proliferative disease of myeloid blasts of the hematopoietic system. It is a highly heterogeneous disease group that can be transformed by malignant transformation of hematopoietic progenitor cells at different stages of normal myeloid cell differentiation and development. In 1976, AML was first divided into 7 types, namely AML-M1 to AML-M7. In 1991, according to the characteristics of immune markers of cells, the diagnosis of AML-M0 was added. [0003] At present, great progress has been made in the treatment of AML, but chemotherapy is still difficult to cure patients, and most patients will still relapse ...
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IPC IPC(8): G01N33/574
CPCG01N33/57426G01N33/56966
Inventor 刘兴娟王绪华刘丽媛秦晓明王显凤吴纯斌梁超陈忠黄士昂
Owner 北京海思特医学检验实验室有限公司
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