Multi-PCR primer and method for constructing leukemia minimal residual disease TCR library based on high-throughput sequencing

Abstract

The invention provides a multi-PCR primer for constructing a leukemia minimal residual disease TCR library based on high-throughput sequencing; the primer comprises a forward primer and a reverse primer, wherein the forward primer is a forward primer group consisting of nucleotide sequences which are 0-3 bases more than or less than nucleotide sequences as shown in SEQ ID NO: 1-SEQ ID NO: 25; and the reverse primer consists of nucleotide sequences which are 0-3 bases more than or less than nucleotide sequences as shown in SEQ ID NO: 26-SEQ ID NO: 28. The multi-PCR primer group provided by the invention can be used for effectively constructing a T lymphocyte receptor high-throughput sequencing library of patients of leukemia, and abundant leukemia minimal residual disease TCR rearrangement information can be obtained.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a multiplex PCR primer and a method for constructing a TCR 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, reg...

AI Technical Summary

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

Images