血浆游离DNA全基因组甲基化测序的实用稳定性评估

Practical stability of whole-genome bisulfite sequencing using plasma cell-free DNA

摘要:

随着液体活检技术的发展，血浆游离DNA成为当前的研究热点之一。血浆游离DNA的全基因组甲基化测序被认为在癌症检测等医学应用拥有巨大潜力，但目前尚缺乏针对该实验流程的实用稳定性评估。文中利用两名志愿者在不同时间采样的血浆游离DNA，在不同实验平台分别进行DNA甲基化的重亚硫酸盐转化前建库 (Pre-BS)、转化后建库 (Post-BS) 和常规DNA建库，获取多因素影响下的测序数据样本。在此基础上，建立了一套血浆游离DNA测序数据分析的质量控制参考流程，综合评估了血液采集提取、游离DNA建库测序过程的实用稳定性，为血浆游离DNA全基因组甲基化测序应用于临床液体活检提供实用性的基础参考。

Abstract:

With the development of liquid biopsy technology, plasma cell-free DNA (cfDNA) becomes one of the research hotspots. Whole-genome bisulfite sequencing of plasma cell-free DNA has shown great potential medical applications such as cancer detection. However, the practical stability evaluation is still lacking. In this study, plasma cell-free DNA samples from two volunteers at different time were collected and prepared for sequencing in multiple laboratories. The library preparation strategies include pre-bisulfite, post-bisulfite and regular whole-genome sequencing. We established a set of quality control references for plasma cell-free DNA sequencing data and evaluated practical stability of blood collection, DNA extraction, and library preparation and sequencing depth. This work provided a technical practice guide for the application of plasma cfDNA methylation sequencing for clinical applications.