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Reagent kit for detecting lymphoma genovariation and application of reagent kit

A gene mutation and lymphoma technology, applied in the detection kit for lymphoma gene mutation, lymphoma gene mutation detection and DLBCL cell origin typing, can solve the problems of inability to accurately identify DLBCL, inconsistency, poor reproducibility, etc.

Active Publication Date: 2020-09-15
BEIJING GENEPLUS TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the limitations of IHC itself, IHC cannot accurately identify 10-15% of DLBCL that cannot be classified, and the reproducibility is poor, and there are also inconsistent phenomena in predicting prognosis [Swerdlow, S.H., Campo, E., Pileri, S.A., Harris, N.L., Stein, H., & Siebert, R., et al. (2016). The 2016 revision of the world health organization classification of lymphoid neoplasms. Blood, 127(20), 2375-2390.], Special However, with the emergence of rituximab, the prognostic value of COO typing by IHC technology has been continuously challenged, and many studies have even shown contradictory results
[0005] According to the review, GEP is the "gold standard" of COO typing, but the sample requirements are high, which limits its clinical use; the typing method based on gene expression has a high consistency with "GEP", and solves the problem of Experimental sampling (fresh tissue) is limited, but it can only be applied to tissue samples, and the cost is high

Method used

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  • Reagent kit for detecting lymphoma genovariation and application of reagent kit
  • Reagent kit for detecting lymphoma genovariation and application of reagent kit
  • Reagent kit for detecting lymphoma genovariation and application of reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] Example 1 Probe set for detection of lymphoma gene variation

[0091] 1. Probe Design

[0092] A total of 413 high-frequency mutation genes, driver mutation genes, targeted drug-related genes, and development-related genes of common lymphoma subtypes were selected. Each gene is covered by multiple probes, among which the probes of 399 genes cover the entire protein coding (CDS) region (see Table 1); the probes of 8 genes only cover the coding regions of some common mutations or Promoter regions (Table 2), probe coverage was performed on the non-coding regions of 26 genes (of which 5 genes only covered non-coding regions), and the main purpose was to detect gene fusions common in lymphoma (Table 3). Among them, BCL2, BCL6 and MYC genes are the three most frequently fused genes in lymphoma. This probe has fully covered the breakpoint region, and carried out a targeted design for the common fusion region of its main partner gene IGH, to To ensure detection sensitivity, t...

Embodiment 2

[0105] Example 2. Clinical application of probe sets in different lymphoma patients

[0106] In this example, 1 case of DLBCL, 1 case of high-grade B-cell lymphoma, 1 case of chronic lymphocytic leukemia, 1 case of extranodal NK / T-cell lymphoma nasal type and 1 case of nodular sclerosis Hodgkin lymphoma Tissue or plasma samples are tested for gene variation to illustrate the value of the present invention in clinical application of different lymphoma patients.

[0107] The implementation process of the present invention is as figure 1 .

[0108] 1. Sample processing and DNA extraction

[0109] The scope of application of samples includes surgically resected fresh pathological tissues, formaldehyde-fixed paraffin-embedded case tissues, paraffin sections, bone marrow or plasma. The sequencing results of buccal swabs / granulocytes were used as controls.

[0110] 1.1 Separation of plasma and granulocytes:

[0111] 1) Draw 10mL of peripheral blood from each person and put it in...

Embodiment 3

[0193] Example 3. The detection ability of the probe set for common gene fusions in lymphoma

[0194] In this example, 3 cases of lymphoma positive fusion standard products (numbers LY-7, LY-8 and JEK0-1) and 5 cases of clinical FISH fusion tests were positive (numbers P002, P006, P007, P008 and P009) Or IHC positive lymphoma clinical samples for gene fusion detection. The specific detection method is shown in Example 2, and the obtained result is the NGS detection result. Compare the consistency of NGS test results with common clinical methods.

[0195] NGS test results:

[0196] The 3 cases of positive standards involved a total of 5 fusions, and all fusion variants were detected, with a sensitivity of 100%. In addition, variants detected by NGS can clarify the location of fusion breakpoints. The specific detection results are shown in the table below:

[0197] Table 14 NGS detection of gene fusion results

[0198]

[0199] Note: 8:128748095 means position 128748095...

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Abstract

The invention discloses a reagent kit for detecting lymphoma genovariation and an application of the reagent kit, and particularly discloses the reagent kit for detection or auxiliary detection of variation of a lymphoma related gene. The reagent kit comprises a substance for detecting BCL2,BCL6, MYC and / or gene IGH fusion, the substance is a complete set of DNA probes, and the complete set of theDNA probes comprises 376 probes as shown in SEQID NO:1 to SEQ ID NO:376. The reagent kit can be used for detection or auxiliary detection of the variation of the lymphoma related gene, substyping ofcells of origin (cell of origin, COO) of patients suffering from diffuse large B-cell lymphoma, and auxiliary diagnosis, prognostication judging and / or targeted drug prediction of the patients suffering from lymphoma.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for detecting lymphoma gene variation. It particularly relates to the detection of lymphoma gene variation and the origin typing of DLBCL cells by using the kit. Background technique [0002] Lymphoma is a malignant tumor originating from the lymphatic hematopoietic system, ranking the eighth among common malignant tumors. In recent years, lymphoma has become the fastest-growing malignant hematological tumor in my country. There are many kinds of them, and their morphology, biological behavior and clinicopathological features are heterogeneous, which has caused great troubles to the diagnosis and treatment of pathologists and clinicians. [0003] Diffuse large B-cell lymphoma (DLBCL) is a malignant tumor originating from B lymphocytes and is the most common non-Hodgkin's lymphoma. In my country, DLBCL accounts for about 40% of all non-Hodgkin's lymphoma [Yang, Q.P., Zhang, W...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6858C12N15/11G16B20/20G16B20/50G16B15/30
CPCC12Q1/6886C12Q1/6858G16B20/20G16B20/50G16B15/30C12Q2600/106C12Q2600/118C12Q2600/112C12Q2600/156C12Q2535/122C12Q2537/165
Inventor 杨玲易鑫管彦芳易玉婷杜新华刘涛徐亚平李盼松
Owner BEIJING GENEPLUS TECH
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