Method for detecting minimal residual disease of T cell leukemia

Abstract

The invention discloses a method for detecting the minimal residual disease of T cell leukemia. The method comprises a multi-PCR primer, wherein the multi-PCR primer comprises an upstream primer and a downstream primer; the upstream primer is an upstream primer group formed by sequences of SEQ ID No:81 to SEQ ID No:112; the downstream primer comprises a nucleotide sequence of SEQ ID No:114. The detection method comprises the steps of (S1) extracting a human peripheral blood mononuclear cell to obtain a total RNA; (S2) carrying out multi-PCR reaction on the obtained RNA by adopting a one-step RT-PCR method to obtain a multi-PCR product; (S3) after multi-PCR reaction is completed, purifying the PCR product by magnetic beads and removing excessive primers; (S4) adding a basic group A to a 3' end of the purified multi-PCR product and obtaining a PCR product with a 3' cohesive end A; and (S5) connecting the PCR product with the 3' cohesive end A and a linker sequence to obtain a connection product containing a linker. According to the method for detecting the minimal residual disease of the T cell leukemia, the detection efficiency is improved and the method is suitable for popularization.

Description

Technical field [0001] The present invention relates to the field of molecular biology technology, in particular to a method for detecting minimal residual disease of T cell leukemia. Background technique [0002] Acute lymphocytic leukemia (ALL) is one of the most common adult acute leukemias. It is a type of malignant hematological disease characterized by the malignant proliferation, accumulation and infiltration of proto-proliferative lymphocytes. A number of studies have reported that the complete remission rate (CR) of ALL patients is 70%-90%, and the 3-5 year disease-free survival rate is 60%. Although the initial CR rate of adult ALL has increased significantly, the recurrence rate is high and the long-term survival rate is still low. Similarly, childhood acute myeloid leukemia (AML) accounts for about 20% of childhood acute leukemia (AML), but it accounts for more than 50% of childhood deaths from AL. The current advanced treatment schemes using new drugs and hematopoi...

AI Technical Summary

Problems solved by technology

Although PCR has high sensitivity, there are few fusion genes that can be used for PCR monitoring, and the scope of use is narrow; although gene mutations and proto-oncogene overexpression have high coverage, the sensitivity is low; FCM is suitable for most patients, and the results are accurate, fast, and efficient. Low cost, high sensitivity, up to 10 -3 ~10 -4

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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