Zhou Yang, Yang Gongda, Liu Haitao, Chen Yawen, Li Xiaofeng, Ge Jun, Wang Xiaochen, Niu Haishan, Dong Wenzhao, Jiang Tingya, Shi Haifeng, Peng Longkai, Liu Lei
Institute of Life Sciences, Jiangsu University, Zhenjiang, China.
Institute of Life Sciences, Jiangsu University, Zhenjiang, China; AlloDx Biotech Co, Ltd, Suzhou, China.
Transplant Proc. 2019 Jul-Aug;51(6):1699-1705. doi: 10.1016/j.transproceed.2019.04.051.
Rejection and infection are 2 major complications affecting the health and survival of patients receiving an allograft organ transplantation. We describe a diagnostic assay that simultaneously monitors for rejection and infection in recipients of kidney transplant by sequencing of cell-free DNA (cfDNA) in plasma.
By using cfDNA in plasma, we established a noninvasive method that simultaneously monitors rejection and infection in patients with a history of organ transplant. A total of 6200 single-nucleotide polymorphisms were captured by liquid hybridization and sequenced by next-generation sequencing. The donor-derived cfDNA (ddcfDNA) level was calculated based on maximum likelihood estimation, without relying on the donor's genotype. We also analyzed the nonhuman cfDNA to test for infections in the patients' plasma.
Artificial ddcfDNA levels quantified by a donor-dependent and donor-independent algorithm were significantly correlated, with the multivariate coefficient of determination, or R value, of 0.999. This technique was applied on 30 patients (32 samples) after kidney transplantation, and a significant difference was observed on the ddcfdNA levels between nonrejection and rejection. Furthermore, 1 BK virus infection and 1 cytomegalovirus infection were revealed by this method, and the enrichment efficiency of the viral sequences was 114 and 489 times, respectively, which are consistent with clinical results.
This method can be used to simultaneously monitor for acute rejection as well as a broad spectrum of infections for patients of allograft organ transplant because it provides comprehensive information for clinicians to optimize immunosuppression therapy.
排斥反应和感染是影响接受同种异体器官移植患者健康和生存的两大主要并发症。我们描述了一种诊断检测方法，通过对血浆中游离DNA（cfDNA）进行测序，同时监测肾移植受者的排斥反应和感染情况。
通过使用血浆中的cfDNA，我们建立了一种非侵入性方法，用于同时监测有器官移植史患者的排斥反应和感染情况。通过液相杂交捕获总共6200个单核苷酸多态性，并通过下一代测序进行测序。基于最大似然估计计算供体来源的cfDNA（ddcfDNA）水平，而不依赖于供体的基因型。我们还分析了非人类cfDNA以检测患者血浆中的感染情况。
通过依赖供体和不依赖供体的算法量化的人工ddcfDNA水平显著相关，多元决定系数或R值为0.999。该技术应用于30例肾移植术后患者（32个样本），在非排斥和排斥状态下的ddcfDNA水平上观察到显著差异。此外，该方法还检测出1例BK病毒感染和1例巨细胞病毒感染，病毒序列的富集效率分别为114倍和489倍，与临床结果一致。
该方法可用于同时监测同种异体器官移植患者的急性排斥反应以及广泛的感染情况，因为它为临床医生优化免疫抑制治疗提供了全面信息。
