Method for detecting MLL-AF4 fusion gene

Abstract

The invention discloses a method for detecting an MLL-AF4 fusion gene. The method sequentially comprises the following steps: 1, acquiring sample DNA, 2, preparing a PCR reaction liquid, wherein a primer used for amplifying the MLL-AF4 fusion gene is F: 5'-CCGCCCAAGTATCCCTGTAAAC-3', and a primer used for amplifying the MLL-AF4 fusion gene is R: 3'-TCGTTATGTATGCGGATCATGTCG-5', 3, carrying out target strip amplification by taking the DNA of a to-be-detected sample as a template, and amplifying a required DNA fragment through the designed primers, 4, carrying out target DNA recovery by carrying out 1% agarose gel electrophoresis on the PCR reaction solution, and recovering a target strip, 5, purifying the DNA fragments obtained in the step 1, 6, carrying out conversion on the purified DNA, and 7, carrying out sequencing, and judging whether the fusion gene is the MLL-AF4 fusion gene or not. According to the method for detecting the MLL-AF4 fusion gene, primers suitable for PCR amplification are designed, high specificity, accuracy and sensitivity are achieved in the detection process, a target band is amplified more rapidly and efficiently, and repeatability is achieved.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a method for detecting MLL-AF4 fusion gene. Background technique [0002] MLL (mixed lineage leukemia) gene abnormality is a very common chromosomal abnormality in acute leukemia. Most acute leukemia patients with MLL gene abnormality have a poor prognosis, which is caused by t(4;11)(q21;q23) translocation The formed MLL-AF4 fusion gene is the second most common chromosomal abnormality in acute lymphoblastic leukemia. Most patients are resistant to conventional chemotherapy, and they are prone to relapse even after allogeneic hematopoietic stem cell transplantation, which is an independent poor prognosis factor. [0003] Among the molecular biology methods widely used at home and abroad to detect the MLL-AF4 fusion gene, although the results of fluorescence in situ hybridization (FISH) are relatively intuitive, they can only perform qualitative detection ...

AI Technical Summary

Problems solved by technology

[0002] MLL (mixed lineage leukemia) gene abnormality is a very common chromosomal abnormality in acute leukemia. Most acute leukemia patients with MLL gene abnormality have a poor prognosis, which is caused by t(4;11)(q21;q23) translocation The formed MLL-AF4 fusion gene is the second most common chromosomal abnormality in acute lymphoblastic leukemia, most patients are resistant to conventional chemotherapy, and are prone to relapse even after allogeneic hematopoietic stem cell transplantation, which is an independent poor prognosis factor

[0003] Among the molecular biology methods widely used at home and abroad to detect the MLL-AF4 fusion gene, although the results of fluorescence in situ hybridization (FISH) are relatively intuitive, they can only perform qualitative detection and the operation is complicated, requiring large-scale equipment such as fluorescence microscopes and The diagnostic opinions of pathologists have strict requirements on testing institutions, which is not conducive to the rapid detection of clinical samples

Traditional solid-phase biochip (Biochip) or gene chip technology has outstanding weaknesses such as poor repeatability, insufficient sensitivity, cumbersome operation, and high cost