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

作　　者：方欢 钟碧溪 魏磊[1] 张祥林 张威[1] 汪小我[1] Huan Fang;Bixi Zhong;Lei Wei;Xianglin Zhang;Wei Zhang;Xiaowo Wang(Bioinformatics Division,BNRIST,Ministry of Education Key Laboratory of Bioinformatics,Center for Synthetic and Systems Biology,Department of Automation,Tsinghua University,Beijing 100084,China)

机构地区：[1]清华大学自动化系合成与系统生物学研究中心生物信息学教育部重点实验室北京信息科学与技术国家研究中心生物信息学研究部

出　　处：《生物工程学报》2019年第12期2284-2294,共11页Chinese Journal of Biotechnology

基　　金：国家自然科学基金(No.61721003)资助~~

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。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.

关 键 词：血浆游离DNA 全基因组 DNA甲基化 片段化模式 微量DNA建库 

分 类 号：R440[医药卫生—诊断学]