Fang Huan, Zhong Bixi, Wei Lei, Zhang Xianglin, Zhang Wei, Wang Xiaowo
Bioinformatics Division, BNRIST, Ministry of Education Key Laboratory of Bioinformatics, Center for Synthetic and Systems Biology, Department of Automation, Tsinghua University, Beijing 100084, China.
Sheng Wu Gong Cheng Xue Bao. 2019 Dec 25;35(12):2284-2294. doi: 10.13345/j.cjb.190281.
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.
随着液体活检技术的发展，血浆游离DNA（cfDNA）成为研究热点之一。血浆游离DNA的全基因组亚硫酸氢盐测序已显示出在癌症检测等医学应用方面的巨大潜力。然而，目前仍缺乏实际稳定性评估。在本研究中，收集了两名志愿者不同时间的血浆游离DNA样本，并在多个实验室进行测序准备。文库制备策略包括亚硫酸氢盐处理前、处理后及常规全基因组测序。我们建立了一套血浆游离DNA测序数据的质量控制参考标准，并评估了采血、DNA提取、文库制备及测序深度的实际稳定性。这项工作为血浆cfDNA甲基化测序在临床应用中的应用提供了技术实践指南。
