Preparation of quality control reference strain for myeloid leukemia fusion gene detection

Abstract

The invention relates to preparation of a quality control reference strain for myeloid leukemia fusion gene detection and belongs to the medical field. The preparation is characterized in that a recombinant human granulocyte colony-stimulating factor is introduced and a marrow cell fusion pre-culture step taking the granulocyte colony-stimulating factor as an inducer is added to promote directional differentiation, increment and maturity of cells in stages of grains, so that the fusion rate of the grains is increased. The granulocyte colony-stimulating factor takes part in infusion of marrow cells, screening of hybrid cells and full-course induction of cloning passage amplification thereof, so that differentiation of the cells in stages of grains is facilitated, therefore, the quality control reference strain for myeloid leukemia fusion gene detection amplified indefinitely in vitro obtained by fusing human marrow cells and mouse marrow cells is prepared. Verified by the infusion gene,the strain is applied to quality control for myeloid leukemia fusion gene detection.

Description

technical field [0001] The invention relates to the preparation of a myeloid leukemia fusion gene detection quality control reference strain in the medical field, which is mainly used for quality control of acute and chronic myeloid leukemia fusion gene detection methods. 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 leukemia (CML) Genes,...

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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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