Leukemia B cell minimal residual detection method

Abstract

The invention relates to the technical field of biological detection, in particular to a leukemia B cell minimal residual detection method. According to the invention, 10 upstream primer sequences are designed directed at a variable V region of human PCR, 1 downstream primer sequence is designed respectively directed at A, D, M, G gene constant regions of BCR, 2 downstream primer sequences are designed directed at an E gene constant region of BCR, and multiplex PCR is carried out to amplify a target sequence, after purification of the target sequence, a basic group A is added to the 3' end of the product sequence and the product is connected to an adapter sequence, the connection product containing the adapter is subjected to PCR amplification with sequencing primers so as to obtain an amplification product, and then purification and sequencing are carried out, and the sequencing result is compared with a cancer cell RNA amplification product sequence. The method has sensitivity up to 10<-6>, and wide detection coverage, and can detect almost all B cell receptors.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a method for detecting minimal residues of leukemia B cells. Background technique [0002] Acute lymphoblastic 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 prolymphocytes. A number of studies have reported that the complete remission rate (CR) of ALL patients reaches 70% to 90% and the 3-5 year disease-free survival rate is 60% with systemic treatment. Although the initial CR rate of adult ALL has been significantly improved, 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 accounts for more than 50% of children's AL deaths. At present, advanced treatment programs using new drugs and hem...

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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