Perspective: sensitive detection of residual lymphoproliferative disease by NGS and clonal rearrangements—how low can you go?

Marcus H Hansen,Oriane Cédile,Thomas S Larsen,Niels Abildgaard,Charlotte G Nyvold

doi:10.1016/j.exphem.2021.03.005

Marcus H Hansen, Oriane Cédile + Show 3 more

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https://doi.org/10.1016/j.exphem.2021.03.005

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Malignant lymphoproliferative disorders collectively constitute a large fraction of the hematological cancers, ranging from indolent to highly aggressive neoplasms. Being a diagnostically important hallmark, clonal gene rearrangements of the immunoglobulins enable the detection of residual disease in the clinical course of patients down to a minute fraction of malignant cells. The introduction of next-generation sequencing (NGS) has provided unprecedented assay specificity, with a sensitivity matching that of polymerase chain reaction-based measurable residual disease (MRD) detection down to the 10-6 level. Although reaching 10-6 to 10-7 is theoretically feasible, employing a sufficient amount of DNA and sequencing coverage is placed in the perspective of the practical challenges when relying on clinical samples in contrast to controlled serial dilutions. As we discuss, the randomness of subsampling must be taken into account to accommodate the sensitivity threshold-in terms of both the required number of cells and sequencing coverage. As a substantial part of the reviewed studies do not state the depth of coverage or even amount of DNA in some cases, we call for increased transparency to enable critical assessment of the MRD assays for clinical implementation and feasibility.

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Constituting a key component of the adaptive immune system, the B lymphocytes are highly specialized in antigen presentation, BCR antigen binding, and
Detection of residual lymphoproliferative disease It has long been recognized that measurable residual disease (MRD) during remission is of prognostic value, there is a strong incentive for pushing the limit of detection further to detect even smaller amounts of clonal cells than possible with current qPCR techniques
Thompson et al [9] reported that the majority of patients found MRD negative by flow cytometry had measurable disease when evaluated with next-generation sequencing (NGS) down to the 10−6 level, but they noted that the recent developments in flow cytometry may provide a resolution comparable to that of NGS

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Introduction

Constituting a key component of the adaptive immune system, the B lymphocytes are highly specialized in antigen presentation, BCR antigen binding, and. We address the theoretical amount of DNA and sequencing reads needed to confidently detect MRD at a given sensitivity level and factors that may influence the resolution.

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