Kim Jin Ju, Kim Soo-Jeong, Lim Seoyoung, Lee Seung-Tae, Choi Jong Rak, Shin Saeam, Hwang Doh Yu
Department of Laboratory Medicine, Yonsei University College of Medicine, Yongin Severance Hospital, Yongin, Korea.
Department of Internal Medicine, Division of Hemato-Oncology, Yonsei University College of Medicine, Yongin Severance Hospital, Yongin, Korea.
Cancer Cell Int. 2024 Aug 12;24(1):282. doi: 10.1186/s12935-024-03470-7.
Risk stratification in multiple myeloma (MM) patients is crucial, and molecular genetic studies play a significant role in achieving this objective. Enrichment of plasma cells for next-generation sequencing (NGS) analysis has been employed to enhance detection sensitivity. However, these methods often come with limitations, such as high costs and low throughput. In this study, we explore the use of an error-corrected ultrasensitive NGS assay called positional indexing sequencing (PiSeq-MM). This assay can detect somatic mutations in MM patients without relying on plasma cell enrichment.
Diagnostic bone marrow aspirates (BMAs) and blood samples from 14 MM patients were used for exploratory and validation sets.
PiSeq-MM successfully detected somatic mutations in all BMAs, outperforming conventional NGS using plasma cells. It also identified 38 low-frequency mutations that were missed by conventional NGS, enhancing detection sensitivity below the 5% analytical threshold. When tested in an actual clinical environment, plasma cell enrichment failed in most BMAs (14/16), but the PiSeq-MM enabled mutation detection in all BMAs. There was concordance between PiSeq-MM using BMAs and ctDNA analysis in paired blood samples.
This research provides valuable insights into the genetic landscape of MM and highlights the advantages of error-corrected NGS for detecting low-frequency mutations. Although the current standard method for mutation analysis is plasma cell-enriched BMAs, total BMA or ctDNA testing with error correction is a viable alternative when plasma cell enrichment is not feasible.
多发性骨髓瘤（MM）患者的风险分层至关重要，分子遗传学研究在实现这一目标中发挥着重要作用。为进行下一代测序（NGS）分析而富集浆细胞已被用于提高检测灵敏度。然而，这些方法往往存在局限性，如成本高和通量低。在本研究中，我们探索使用一种名为位置索引测序（PiSeq-MM）的纠错超灵敏NGS检测方法。该检测方法可不依赖浆细胞富集来检测MM患者的体细胞突变。
使用14例MM患者的诊断性骨髓穿刺液（BMA）和血液样本作为探索性和验证性数据集。
PiSeq-MM成功检测到了所有BMA中的体细胞突变，优于使用浆细胞的传统NGS。它还识别出了38个传统NGS遗漏的低频突变，提高了低于5%分析阈值的检测灵敏度。在实际临床环境中进行测试时，大多数BMA（14/16）的浆细胞富集失败，但PiSeq-MM能够在所有BMA中检测到突变。使用BMA的PiSeq-MM与配对血液样本中的ctDNA分析结果具有一致性。
本研究为MM的遗传图谱提供了有价值的见解，并突出了纠错NGS在检测低频突变方面的优势。尽管目前突变分析的标准方法是富集浆细胞的BMA，但当浆细胞富集不可行时，全BMA或经纠错的ctDNA检测是一种可行的替代方法。
