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Kit for detecting gene mutation of diffuse large B-cell lymphoma

A detection kit and a diffuse technology, applied in the field of gene mutation detection kits for diffuse large B-cell lymphoma, can solve problems such as sensitivity and poor prognosis

Pending Publication Date: 2019-07-26
珠海铂华生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the Y196 mutation of the CD79B gene is common in ABC-type diffuse large B-cell lymphoma, and the prognosis of this subtype is poor. Patients with this mutation may be sensitive to ibrutinib, fostamatinib and other drugs

Method used

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  • Kit for detecting gene mutation of diffuse large B-cell lymphoma
  • Kit for detecting gene mutation of diffuse large B-cell lymphoma
  • Kit for detecting gene mutation of diffuse large B-cell lymphoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Collection of samples and arrangement of sample data: The applicant collected formalin-fixed paraffin-embedded (FFPE) section samples from 118 DLBCL patients in China, and each sample took a patient's oral swab as an internal control of the patient. The sample collection process meets the following conditions:

[0036](1) All patient samples are samples from patients diagnosed as diffuse large B-cell lymphoma through pathological sections, karyotype analysis, immunology, flow cytometry and other methods;

[0037] (2) All patient samples come from lymph node samples collected at the first visit and oral swab control samples collected within 1 month;

[0038] (3) All the samples were matched with relevant clinical data collected.

[0039] Based on the samples collected above, DNA was extracted using a commercial DNA extraction kit (Qiagen, "GeneReadDNA FFPE KIT"), and whole exome sequencing was performed using the kit xGen Exome Research Panel v1.0 (Cat. No.: 1056115) ,...

Embodiment 2

[0043] Based on the relationship between the G185A mutation of the KCNG1 gene and DLBCL found in the present invention, the present invention provides a DLBCL gene mutation detection kit based on high-throughput sequencing technology. KCNG1 gene, MYD88, CD79B, EZH2, B2M, NOTCH2, STAT6, TP53 genes reported in the prior art. The detection range of the kit is the entire exon region of the above-mentioned gene, and its main components include the primers in the above-mentioned Table 1. The kit is based on high-throughput sequencing technology, constructs a sequencing library by multiplex PCR, and performs sequencing on the library and data sequencing to detect whether there are mutations in the above genes. The specific implementation process is as follows:

[0044] (1) Extraction of sample DNA: Genomic DNA was extracted from FFPE samples using a commercial DNA extraction kit (Qiagen, "GeneRead DNAFFPE KIT"), and the quality of DNA 260 / 280 was determined using Nonodrop, and the D...

Embodiment 3

[0092]Select 2 cases of KCNG1 gene G185A mutation samples in Example 1, and another 20 cases of DLBCL formalin-fixed paraffin-embedded section (FFPE section) samples, 10 cases of normal human peripheral blood samples, using the kit in Example 2 , according to its implementation process for detection, the detection results are as follows:

[0093] Table 2 Quality control table of kit off-machine data

[0094]

[0095]

[0096] The sample number starting with N indicates a normal human sample, the beginning of D indicates a DLBCL sample, and the beginning of E indicates a sample of the KCNG1 gene G185A mutation obtained by whole exome sequencing in Example 1, the sequencing quality is >30, the depth is >500, and the uniformity The sequencing results of >80% were regarded as qualified sample sequencing results, and Table 2 shows that all the above-mentioned samples showed qualified sequencing results.

[0097] In this experiment, the mutation results of all kit-related gen...

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Abstract

The invention discloses a kit for detecting gene mutation of diffuse large B-cell lymphoma and belongs to the field of gene detection and disease diagnosis. It is found that a certain connection exists between KCNG1 gene and diffuse large B cell lymphoma (DLBCL), and the kit can be used for auxiliary diagnosis of DLBCL by detecting whether G185A mutation exists in KCNG1 gene or not. The genes detected by the kit for detecting DLBCL gene mutation include a KCNG1 gene and MYD88, CD79B, EZH2, B2M, NOTCH2, STAT6 and TP53 genes which are determined to be related to DLBCL generation in the prior art. The invention provides a new material for auxiliary diagnosis of DLBCL; the kit can detect the mutation condition of the gene at one time, and provides a basis for early diagnosis, typing judgment,prognosis stratification and medication guidance of DLBCL.

Description

technical field [0001] The invention belongs to the field of gene detection and disease diagnosis, and in particular relates to a diffuse large B-cell lymphoma gene mutation detection kit. Background technique [0002] Diffuse large B-cell lymphoma (DLBCL) is a common type of non-Hodgkin's lymphoma, accounting for about 50% of non-Hodgkin's lymphoma and 30% of the overall incidence of lymphoma. DLBCL was first defined in 1994, and its new subtypes were continuously added in subsequent World Health Organization (WHO) guidelines. In the 2008 WHO guidelines, DLBCL was defined as a diffusely proliferating large B-cell malignant tumor with tumor cell nuclei larger than or equal to normal phagocytic cell nuclei, or twice as large as normal lymphocytes. DLBCL shows certain heterogeneity clinically, and is characterized by a rapidly increasing painless mass. The tumor mainly originates in the lymph nodes, but about 30% of the patients develop outside the lymph nodes, generally show...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6858C40B50/06
CPCC12Q1/6888C12Q1/6858C40B50/06C12Q2600/106C12Q2600/112C12Q2600/118C12Q2600/156C12Q2535/122C12Q2531/113C12Q2537/165Y02A50/30
Inventor 朱洲杰陈少平卫晓光
Owner 珠海铂华生物工程有限公司
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