Diagnostic Molecular Pathology, Department of Pathology and Laboratory Medicine, Memorial Sloan Kettering Cancer Center, New York, New York; Hematopathology Service, Department of Pathology and Laboratory Medicine, Memorial Sloan Kettering Cancer Center, New York, New York.
Loxo Oncology, Inc., Stamford, Connecticut.
J Mol Diagn. 2024 Mar;26(3):168-178. doi: 10.1016/j.jmoldx.2023.11.009. Epub 2023 Dec 14.
Next-generation sequencing (NGS)-based measurable residual disease (MRD) monitoring in post-treatment settings can be crucial for relapse risk stratification in patients with B-cell and plasma cell neoplasms. Prior studies have focused on validation of various technical aspects of the MRD assays, but more studies are warranted to establish the performance characteristics and enable standardization and broad utilization in routine clinical practice. Here, the authors describe an NGS-based IGH MRD quantification assay, incorporating a spike-in calibrator for monitoring B-cell and plasma cell neoplasms based on their unique IGH rearrangement status. Comparison of MRD status (positive or undetectable) by NGS and flow cytometry (FC) assays showed high concordance (91%, 471/519 cases) and overall good linear correlation in MRD quantitation, particularly for chronic lymphocytic leukemia and B-lymphoblastic leukemia/lymphoma (R = 0.85). Quantitative correlation was lower for plasma cell neoplasms, where underestimation by FC is a known limitation. No significant effects on sequencing efficiency by the spike-in calibrator were observed, with excellent inter- and intra-assay reproducibility within the authors' laboratory, and in comparison to an external laboratory, using the same assay and protocols. Assays performed both at internal and external laboratories showed highly concordant MRD detection (100%) and quantitation (R = 0.97). Overall, this NGS-based MRD assay showed highly reproducible results with quantitation that correlated well with FC MRD assessment, particularly for B-cell neoplasms.
基于下一代测序（NGS）的治疗后微小残留病（MRD）监测对于 B 细胞和浆细胞肿瘤患者的复发风险分层至关重要。先前的研究集中于验证 MRD 检测的各种技术方面，但需要更多的研究来建立性能特征，并使其在常规临床实践中标准化和广泛应用。在这里，作者描述了一种基于 NGS 的 IGH MRD 定量检测方法，该方法纳入了一种内参校准物，用于监测基于其独特 IGH 重排状态的 B 细胞和浆细胞肿瘤。NGS 和流式细胞术（FC）检测的 MRD 状态（阳性或不可检测）比较显示出高度一致性（91%，471/519 例），MRD 定量总体上具有良好的线性相关性，尤其是在慢性淋巴细胞白血病和 B 淋巴母细胞白血病/淋巴瘤中（R=0.85）。对于浆细胞肿瘤，FC 存在低估的已知局限性，因此定量相关性较低。内参校准物对测序效率没有显著影响，在作者实验室内部和与外部实验室的比较中，该检测具有极好的内和间检测重现性，使用相同的检测方法和方案。在内部和外部实验室进行的检测均显示出高度一致的 MRD 检测（100%）和定量（R=0.97）。总体而言，这种基于 NGS 的 MRD 检测方法具有高度可重复的结果，其定量与 FC MRD 评估相关性良好，尤其是在 B 细胞肿瘤中。
