Development and validation of sensitive BCR::ABL1 fusion gene quantitation using next-generation sequencing.

ABSTRACT:

Background

BCR::ABL1 fusion has significant prognostic value and is screened for chronic myeloid leukemia (CML) disease monitoring as a part of routine molecular testing. To overcome the limitations of the current standard real-time quantitative polymerase chain reaction (RQ-PCR), we designed and validated a next-generation sequencing (NGS)-based assay to quantify BCR::ABL1 and ABL1 transcript copy numbers.

Methods

After PCR amplification of the target sequence, deep sequencing was performed using an Illumina Nextseq 550Dx sequencer and in-house-designed bioinformatics pipeline. The Next-generation Quantitative sequencing (NQ-seq) assay was validated for its analytical performance, including precision, linearity, and limit of detection, using serially diluted control materia

SUBMITTER: Lee H 

PROVIDER: S-EPMC10226238 | biostudies-literature | 2023 May

REPOSITORIES: biostudies-literature