Kit and method for detecting gene mutation of acute lymphoblastic leukemia based on multiplex-PCR targeted high-throughput sequencing

Abstract

The invention discloses a kit for detecting gene mutation of acute lymphoblastic leukemia based on multiplex-PCR targeted high-throughput sequencing and a detection method of the kit. The kit is usedfor detecting multiple exon regions of 19 genes related with the acute lymphoblastic leukemia. The kit contains a primer working solution for detecting relevant gene mutation, a PCR mixed solution, aPCR reaction solution, a digestion solution, a joint P5 and a joint P7. The kit is used for carrying out detection based on a multiplex-PCR targeted high-throughput sequencing technique, has the advantages of high accuracy, sensitivity and throughput and can be used for rapidly and efficiently carrying out standardized quantitative detection on 19 relevant genes of acute lymphoblastic leukemia (ALL).

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a multiple PCR-based targeted high-throughput sequencing acute lymphoblastic leukemia gene mutation detection kit and method. Background technique [0002] Leukemia is a kind of highly heterogeneous malignant blood disease caused by abnormal gene of hematopoietic stem or precursor cells, resulting in abnormal cell proliferation, differentiation or apoptosis. Comprehensive analysis of biology. The most common molecular biological abnormalities in leukemia mainly include: gene mutation, fusion gene, gene expression, and gene mutation is mainly point mutation, gene fragment insertion or deletion. [0003] Acute lymphoblastic leukemia (ALL) is a malignant tumor disease that originates from the abnormal proliferation of B or T lineage cells of lymphocytes in the bone marrow. It mainly occurs in children and adults, and its incidence peaks between the ages of 2 and 5. The main...

AI Technical Summary

Problems solved by technology

Compared with the first-generation (Sanger) sequencing technology, the targeted sequencing technology of multiplex PCR has higher sensitivity and can quantitatively detect the gene mutation load rate; compared with the targeted sequencing of capture probes, the targeted sequencing technology of multiplex PCR The technical library construction process is simpler and can provide a more economical and rapid sequencing solution. Most of the real-time fluorescent PCR (RQ-PCR) and digital PCR are for the detection of known mutation sites in the gene, and the primers are detected during the experiment. The cost of needles is high, and the feasibility of multi-gene mutation screening is poor

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