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.