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Diffuse large b-cell lymphoma molecule pathological typing method and use thereof

A technology for B cells and lymphoma, applied in the field of hematology oncology and medicine, can solve the problems of high specimen requirements, complicated operation, and high cost of gene chip detection

Inactive Publication Date: 2009-07-01
RUIJIN HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Gene chip technology has deepened the understanding of the biological characteristics of DLBCL, but because of its high requirements for specimens (fresh specimens, suitable cryopreservation, etc.), complex operations, and high cost of gene chip detection, the cost of using gene chip detection is 7000-9000 yuan per patient, thus restricting its clinical practicability

Method used

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  • Diffuse large b-cell lymphoma molecule pathological typing method and use thereof
  • Diffuse large b-cell lymphoma molecule pathological typing method and use thereof
  • Diffuse large b-cell lymphoma molecule pathological typing method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1. DLBCL specimen collection, processing and detection

[0076] 1. Specimen Collection

[0077] The cases included in this study were 104 newly diagnosed DLBCL patients. All DLBCL specimens were fixed with 10% neutral formaldehyde, routinely dehydrated, embedded in paraffin, stained with hematoxylin-eosin, and diagnosed according to the new WHO classification by doctors specialized in lymphoma research.

[0078] 2. Specimen Processing

[0079] (1) Prepare tissue sections (paraffin sections are dewaxed to water, frozen sections and cell culture sheets are fixed).

[0080] (2) Antigen retrieval (microwave heating method, suitable for paraffin sections) Place the sections in a high-temperature-resistant container and inject the antigen retrieval solution Tris / EDTA (50mM Tris, 2mM EDTA, pH9.0) so that the sections are below the liquid surface. Heat in a microwave oven to keep the temperature of the liquid in the container between 92°C and 98°C for 5 minutes, take...

Embodiment 2

[0081] Example 2. Immunohistochemical detection and result determination of molecular pathological typing of diffuse large B-cell lymphoma

[0082] 1. Rinse the slices with TBS 4-5 times, shake dry, wipe clean, add 20μl 3% H 2 o 2 - Methanol solution, blocked for 30 minutes in a room temperature humid chamber.

[0083] 2. Take out the slices, rinse with TBS 4-5 times, shake dry, and wipe clean. Cover with 20 μl of blocking solution, and incubate for 30 minutes at room temperature in a humid chamber. Absorb the blocking solution with absorbent paper, do not wash, add dropwise the commercially available primary antibody working solution (CD10, purchased from Novocastra, 1:80; Bcl6, purchased from DAKO, 1:10; MUM1, purchased from DAKO , 1:40). Incubate for 1-2 hours at 37°C or overnight at 4°C.

[0084] 3. Rinse TBST 4-5 times, shake dry, and wipe clean. Add 20 μl of secondary antibody (HRP-labeled, ChemMateTMEnVision+ / HRP) dropwise, and incubate for 1-2 hours at room tempe...

Embodiment 3

[0091] Example 3. Molecular classification of patients with diffuse large B-cell lymphoma

[0092] CD10 and Bcl-6 are used as markers of germinal center cell origin, and CD10(+) or CD10(-) / Bcl-6(+) / MUM1(-) are GCBs; CD10(-) / bcl-6(- ), which are non-GCB categories. MUM1 is used as a marker of the origin of post-germinal center cells. If CD10(-) / bcl-6(+), MUM1 is used to classify: MUM1(+) is non-GCB, and MUM1(-) is GCB (see figure 1 and figure 2 ), that is, CD10(-) / bcl-6(+) / MUM1(+) is non-GCB, and CD10(-) / bcl-6(+) / MUM1(-) is GCB.

[0093] According to the molecular typing method, and according to the immunohistochemical test results in the foregoing examples, among the collected 104 cases of newly diagnosed DLBCL patients, 42 cases were classified as GCB type patients, and the remaining 62 cases were non-GCB type patients.

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Abstract

The invention provides a reagent kit for processing prognosis and / or the selection of treatment schemes on diffuse large b-cell lymphomas and an application therefore. The invention is characterized in that the reagent kit comprises: (i) detecting one or more agents expressed by the CD10 protein in a biomass sample; (ii) detecting one or more agents expressed by the Bcl-6 protein in the biomass sample; (iii) detecting one or more agents expressed by the MUM1 protein in the biomass sample. The reagent kit and method can process objective treatment scheme on the patients of diffuse large b-cell lymphomas, thereby improving the prognosis effect of treatment scheme and saving treatment cost.

Description

technical field [0001] The present invention relates to the fields of hematology-oncology and medicine. More specifically, it relates to a molecular typing marker of diffuse large B-cell lymphoma, a detection method and a kit related to the detection method. Background technique [0002] Diffuse large B-cell lymphoma (DLBCL) is the most common non-Hodgkin's lymphoma, accounting for about 40% of newly diagnosed non-Hodgkin's lymphoma each year. The current incidence of non-Hodgkin's lymphoma in my country is about 3-4 / 100,000, and it is increasing at a rate of 2-3% per year, among which diffuse large B-cell lymphoma (DLBCL) is the most common, accounting for about 50% of tumors, therefore, the annual national new cases are about 40,000-50,000. [0003] DLBCL has significant biological heterogeneity, and patients have significant differences in response to treatment. Conventional combined chemotherapy regimens (CHOP, cyclophosphamide, doxorubicin, vincristine and prednisone) ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/53
Inventor 李军民张庆华夏祖光赵维莅沈志祥宋凯
Owner RUIJIN HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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