Preparation of a quality control reference strain for lymphoblastic leukemia fusion gene detection

A technology that combines genes and leukemia, applied in the medical field, can solve problems that affect detection methods, cannot be repeated, and have a high false positive rate

Inactive Publication Date: 2018-12-07
翁炳焕
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, there are many factors that will affect the accuracy of the detection method. Harismendy et al. reported that the false positive rate of different NGS platforms was as high as 7.8%. reported that 26% of the detected mutations were interpreted differently and 11% of the detected mutations were interpreted completely opposite
In 2016, the FDA calibration project (precision FDA) used the operating software of a gene sequencing company to "read" the DNA sequencing results of the same genetic test sample, and almost 50% of the reading software could n...

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  • Preparation of a quality control reference strain for lymphoblastic leukemia fusion gene detection
  • Preparation of a quality control reference strain for lymphoblastic leukemia fusion gene detection

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Embodiment Construction

[0013] figure 1 It is a hybrid cell clone diagram prepared by the present invention.

[0014] exist figure 1 In the medium, the bone marrow cells of patients with lymphoid leukemia are induced by B lymphocyte stimulating factors to differentiate lymphocytes of various stages. Under the action of PEG, after fusion with mouse myeloma cells, they are screened by HAT for 1 to 2 weeks, and then viewed under an inverted microscope. Shoot (40X) to obtain a hybrid cell clone map.

[0015] Combine below figure 1 , the embodiment of the present invention is described in detail.

[0016] 1. Cells to be fused

[0017] (1) Bone marrow cells to be fused: Bone marrow cells from patients with lymphocytic leukemia who were diagnosed and left after bone marrow cytology examination, containing children's EL / AML1 (46%), E2A / PBX1 (20%), Fusion genes such as BCR / ABLp190, BCR / ABLp210 and / or MLL / AF4, or fusion genes such as M-bcr / abl and / or m-bcr / abl in adults.

[0018] (2) Myeloma cells: mouse...

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Abstract

Preparation of a quality control reference strain for lymphoblastic leukemia fusion gene detection for the medical field is disclosed. The preparation is characterized in that a B lymphocyte stimulator is introduced, and a step of culture before bone marrow cell fusion is additionally arranged with the B lymphocyte stimulator being adopted as an inducer, so as to promote directional differentiation, increase and maturation of lymphocyte-line cells of different stages, thus increasing the fusion rate of the lymphocyte line; the B lymphocyte stimulator participates induction of bone marrow cellfusion, hybrid cell screening and cloning, passage and amplification, thus facilitating differentiation lymphocytes of different stages; accordingly, the reference strain prepared from human bone marrow cells and mouse myeloma cells and capable of in-vitro unlimited amplification is prepared; and through verification by the fusion gene, the reference strain can be applied for quality control in lymphoblastic leukemia fusion gene detection.

Description

technical field [0001] The invention relates to the preparation of a quality control reference strain for detection of lymphoid leukemia fusion genes in the medical field, which is mainly used for quality control of detection methods of fusion genes of acute and chronic lymphocytic leukemia. Background technique [0002] Leukemia is a malignant clonal disease of hematopoietic stem cells, divided into acute and chronic. Acute minimally differentiated myeloblastic leukemia, M2 acute partially differentiated myeloblastic leukemia, M3 acute promyelocytic leukemia, M4 acute myelomonocytic leukemia, and M5 acute monocytic leukemia, etc.) , According to the hematopoietic system can be divided into granulocytic (myeloid) leukemia, lymphoid (lymphoid) leukemia and so on. [0003] Leukemia patients produce fusion genes due to translocation or rearrangement of certain chromosomes, such as BCR-ABL (more than 90%) fusion formed by t(9;22)(q34;q11) translocation in chronic myelogenous le...

Claims

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Application Information

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IPC IPC(8): C12N15/08C12N5/077
CPCC12N5/0652C12N15/02
Inventor 翁炳焕苏岚袁武锋杜雨轩王薇应俊
Owner 翁炳焕
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