Immunogenetics Laboratory, Pathology and Laboratory Medicine, Temple University and Hospital, Lewis Katz School of Medicine, 3401 N. Broad St., Office B242, Philadelphia, PA 19140, USA.
Fox Chase Cancer Center Medical Group, Temple Health, Philadelphia, PA 19140, USA.
Int J Mol Sci. 2023 Jul 23;24(14):11814. doi: 10.3390/ijms241411814.
Chimerism monitoring after allogenic Hematopoietic Cell Transplantation (allo-HCT) is critical to determine how well donor cells have engrafted and to detect relapse for early therapeutic intervention. The aim of this study was to establish and detect mixed chimerism and minimal residual disease using Next Generation Sequencing (NGS) testing for the evaluation of engraftment and the detection of early relapse after allo-HCT. Our secondary aim was to compare the data with the existing laboratory method based on Short Tandem Repeat (STR) analysis. One hundred and seventy-four DNA specimens from 46 individuals were assessed using a commercially available kit for NGS, AlloSeq HCT NGS (CareDx), and the STR-PCR assay. The sensitivity, precision, and quantitative accuracy of the assay were determined using artificially created chimeric constructs. The accuracy and linearity of the assays were evaluated in 46 post-transplant HCT samples consisting of 28 levels of mixed chimerism, which ranged from 0.3-99.7%. There was a 100% correlation between NGS and STR-PCR chimerism methods. In addition, 100% accuracy was attained for the two external proficiency testing surveys (ASHI EMO). The limit of detection or sensitivity of the NGS assay in artificially made chimerism mixtures was 0.3%. We conducted a review of all NGS chimerism studies published online, including ours, and concluded that NGS-based chimerism analysis using the AlloSeq HCT assay is a sensitive and accurate method for donor-recipient chimerism quantification and minimal residual disease relapse detection in patients after allo-HCT compared to STR-PCR assay.
异基因造血细胞移植（allo-HCT）后嵌合体监测对于确定供体细胞的植入情况以及检测复发以进行早期治疗干预至关重要。本研究的目的是建立和检测嵌合体和微小残留病，使用下一代测序（NGS）测试评估植入情况，并在 allo-HCT 后检测早期复发。我们的次要目标是将数据与现有的基于短串联重复（STR）分析的实验室方法进行比较。使用商业上可获得的用于 NGS 的试剂盒，即 AlloSeq HCT NGS（CareDx）和 STR-PCR 分析，对来自 46 个人的 174 个 DNA 标本进行了评估。使用人工嵌合构建体确定了该测定的灵敏度、精密度和定量准确性。在由 28 个混合嵌合体水平组成的 46 个移植后 HCT 样本中评估了该测定的准确性和线性，混合嵌合体水平范围为 0.3-99.7%。NGS 和 STR-PCR 嵌合体方法之间存在 100%的相关性。此外，在由 28 个混合嵌合体水平组成的 46 个移植后 HCT 样本中评估了该测定的准确性和线性，混合嵌合体水平范围为 0.3-99.7%。此外，在由 28 个混合嵌合体水平组成的 46 个移植后 HCT 样本中评估了该测定的准确性和线性，混合嵌合体水平范围为 0.3-99.7%。此外，在两个外部能力验证调查（ASHI EMO）中，该测定的准确性达到 100%。在人工嵌合体混合物中，NGS 测定的检测限或灵敏度为 0.3%。我们对所有已在线发表的 NGS 嵌合体研究进行了综述，包括我们自己的研究，并得出结论，与 STR-PCR 分析相比，使用 AlloSeq HCT 测定进行基于 NGS 的嵌合体分析是一种敏感且准确的方法，可用于 allo-HCT 后供受者嵌合体定量和微小残留病复发检测。
