Next-Generation Sequencing for Measurable Residual Disease Assessment in Acute Leukemia

Kovach, Alexandra E.; Raca, Gordana; Bhojwani, Deepa; Wood, Brent L.

doi:10.1016/j.yamp.2021.07.011

Cited by 2 publications

(1 citation statement)

References 128 publications

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“…We demonstrated that NGS could simultaneously determine major transcript types and MR levels with good sensitivity and acceptable precision. Moreover, NGS-based MRD assays can simultaneously perform high-throughput sequencing of multiple genes and bulk samples[ 20 ].…”

Section: Discussionmentioning

confidence: 99%

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

et al. 2023

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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 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.

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Section: Discussionmentioning

confidence: 99%