Lan James H, Zhang Qiuheng
aDepartment of Pathology & Laboratory Medicine, UCLA Immunogenetics Center, Los Angeles, California, USA bUniversity of British Columbia Clinician Investigator Program, Vancouver, British Columbia, Canada.
Curr Opin Organ Transplant. 2015 Aug;20(4):461-7. doi: 10.1097/MOT.0000000000000217.
Next-generation sequencing (NGS) can overcome traditional methodological barriers to facilitate detailed studies of large genomes. Here, we summarize recent NGS-based developments in histocompatibility and transplantation, and highlight the dynamic range of clinical applications achievable on this platform.
Multiple NGS-based protocols have been established to achieve unambiguous human leukocyte antigen genotyping. These methods are presently engaged to serve the high-throughput demand of large bone marrow registries; however, the scalable nature of NGS makes it an equally attractive technology for select applications within solid organ transplantation. Recently, the exquisite sensitivity of NGS has been leveraged to perform noninvasive allograft monitoring by tracking the dynamics of donor-derived cell-free DNA. Further, NGS-based T-cell receptor and immunoglobulin heavy chain repertoire profiling appear to be useful in clarifying disease-specific diagnoses in certain complex allograft pathology; detecting/quantifying minimal residual disease following allogeneic stem cell transplantation; and tracking donor-reactive T cells to understand the mechanism of tolerance in kidney transplant recipients.
NGS is superior to classical Sanger sequencing in its throughput, sensitivity, and the ability to provide phase-defined sequence data. These unique properties allow its broad application to diverse areas in clinical transplantation.
新一代测序（NGS）能够克服传统方法学障碍，推动对大型基因组进行详细研究。在此，我们总结了近期基于NGS在组织相容性和移植领域的进展，并强调了该平台可实现的临床应用动态范围。
已建立多种基于NGS的方案以实现明确的人类白细胞抗原基因分型。目前这些方法用于满足大型骨髓登记处的高通量需求；然而，NGS的可扩展性使其在实体器官移植的特定应用中同样具有吸引力。最近，通过追踪供体来源的游离DNA动态，利用NGS的高灵敏度进行非侵入性同种异体移植物监测。此外，基于NGS的T细胞受体和免疫球蛋白重链库分析在某些复杂的同种异体移植病理学中明确疾病特异性诊断、检测/定量异基因干细胞移植后的微小残留病以及追踪供体反应性T细胞以了解肾移植受者的耐受机制方面似乎很有用。
NGS在通量、灵敏度以及提供相位定义序列数据的能力方面优于经典的桑格测序。这些独特特性使其能够广泛应用于临床移植的各个领域。
