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- W4310690207 abstract "Abstract Background BCR::ABL1 fusion has significant prognostic value and is screened for chronic myeloid leukaemia (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 materials. A comparison with conventional RQ-PCR was performed with 145 clinical samples from 77 patients. Results The limit of detection of the NQ-seq was the molecular response (MR) 5.6 [ BCR::ABL1 0.00028% international scale (IS)]. The NQ-seq exhibited excellent precision and linear range from MR 2.0 to 5.0. The IS value from the NQ-seq was highly correlated with conventional RQ-PCR. Conclusions We conclude that the NQ-seq is an effective tool for monitoring BCR::ABL1 transcripts in CML patients with high sensitivity and reliability. Prospective assessment of the unselected large series is required to validate the clinical impact of this NGS-based monitoring strategy." @default.
- W4310690207 created "2022-12-15" @default.
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- W4310690207 date "2022-12-05" @default.
- W4310690207 modified "2023-09-24" @default.
- W4310690207 title "Development and validation of sensitive BCR::ABL1 fusion gene quantitation using next-generation sequencing" @default.
- W4310690207 doi "https://doi.org/10.21203/rs.3.rs-2328608/v1" @default.
- W4310690207 hasPublicationYear "2022" @default.
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