Multiple PCR primers and method for constructing leukemia minimal residual disease BCR library based on high-flux sequencing

Abstract

The invention provides multiple PCR primers for constructing a leukemia minimal residual disease BCR library based on high-flux sequencing. The multiple PCR primers comprise upstream primers and downstream primers, wherein the upstream primers are an upstream primer group composed of nucleotide sequences which are 0-3 basic groups more or less than the nucleotide sequences shown in the SEQ ID NO: 1-SEQ ID NO: 13, and the downstream primers are composed of nucleotide sequences which are 0-3 basic groups more or less than the nucleotide sequences shown in the SEQ ID NO: 14-SEQ ID NO: 17. The provided multiple PCR primer group can efficiently construct the B lymphocyte receptor high-flux sequencing library of a leukemia patient, so that rich leukemia minimal residual disease BCR rearrangement information can be obtained.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a multiple PCR primer and a method for constructing a BCR library of leukemia minimal residual lesions based on high-throughput sequencing. Background technique [0002] Leukemia is a malignant clonal disease of the blood system characterized by an increase in immature cells in the bone marrow and / or peripheral blood. At the time of first diagnosis, the total number of leukemia cells in the patient's body was about 10 12 , after chemotherapy to achieve morphological complete remission (Complete Remission, CR), the total number of residual leukemia cells is less than 10 9 , this morphological method is difficult to detect, and a small amount of leukemia cells still remain in the body is called minimal residual disease (minimal residual disease, MRD). Residual MRD in the body causes >50% relapse rate in patients with acute T-lymphoblastic leukemia. Therefore, regu...

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Problems solved by technology

Although the detection sensitivity of mpFC for recurrent disease is 10 -4 , but the complex multi-dimensional data depends on the analysis of the experimenter, and the influence of human factors is large, which is not conducive to clinical standardized detection

In addition, the expression levels of leukemia antigens interfered with the detection of mpFC in MRD after chemotherapy

Relying on molecular means can improve the sensitivity of detecting MRD, which can reach 10 -5 However, real-time quantitative PCR needs to design special primers according to patients to amplify the diverse rearranged sequences, which is expensive, labor-intensive, and difficult to form a standardized experimental process

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