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Updated: Dec 30, 2025
Oncogenic Gene Fusion Detection Using Anchored Multiplex Polymerase Chain Reaction Followed by Next Generation Sequencing
Published on: July 5, 2019
Development and application of a rapid molecular method for detection of fusion genes in pediatric leukemia
Qian Chen1, Zheng Hu1, Yongjun Fang2
1Institute of Pediatrics, Children's Hospital of Nanjing Medical University No.72 Guangzhou Road, Nanjing 210008, China.
Objective:
Fusion gene detection is widely used in the diagnosis and treatment of leukemia. This study developed a rapid detection method of eight common pediatric leukemia fusion genes.
Methods:
In this study, one step multiplex RT-PCR assay was developed for the simultaneous detection of eight common leukemia fusion genes, including BCR-ABL, ETV6-RUNX1, MLL-AF4, E2A-PBX1, AML1-ETO, PML-RARα, CBFβ-MYH11 and SIL-TAL1. The single step RT-PCR approach is mediated by universal primers after obtaining total RNA from bone marrow specimens. The size of the amplified fragments were analyzed by capillary electrophoresis assay. A total of 122 patients with positive leukemia fusion genes were tested by real-time PCR.
Results:
Respectively, 21 cases were detected as CBRB-MYH11 fusion gene, 13 cases were detected as SIL-TAL1 fusion gene, 16 cases were detected as ETV6-RUNX1 fusion gene, 16 cases were detected as E2A-PBX1 fusion gene, 15 cases were detected as PML-RARα fusion gene, 14 cases were detected as AML1-ETO fusion gene, 13 cases were detected as MLL-AF4 fusion gene, except for 1 case where no fusion gene was detected.
Conclusion:
This method has a high accuracy and detection rate. Therefore, one step multiplex RT-PCR combined with a capillary electrophoresis analysis system can be used as an important tool for the clinical diagnosis, treatment and prognosis of pediatric leukemia.
