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

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...

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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 not repeat the same results

It is known that the EB virus transfection method can solve the problem of culture and expansion of B lymphocytes. We have also used the EB virus transfection method to establish lymphocyte lines that are easily misdiagnosed with chromosomal abnormalities and use them for quality control of lymphocyte culture and chromosome preparation. Tried and tested again and again, but failed to transfect bone marrow cells with Epstein-Barr virus, it may be because Epstein-Barr virus can only transform mature B lymphocytes but not bone marrow cells

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