Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University and Beijing Key Laboratory of Emerging Infectious Diseases, Beijing 100015, China.
School of Life Sciences, Tsinghua University, Beijing 100084, China.
Mol Cell. 2020 Dec 17;80(6):1123-1134.e4. doi: 10.1016/j.molcel.2020.11.030. Epub 2020 Nov 20.
Analyzing the genome of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from clinical samples is crucial for understanding viral spread and evolution as well as for vaccine development. Existing RNA sequencing methods are demanding on user technique and time and, thus, not ideal for time-sensitive clinical samples; these methods are also not optimized for high performance on viral genomes. We developed a facile, practical, and robust approach for metagenomic and deep viral sequencing from clinical samples. We demonstrate the utility of our approach on pharyngeal, sputum, and stool samples collected from coronavirus disease 2019 (COVID-19) patients, successfully obtaining whole metatranscriptomes and complete high-depth, high-coverage SARS-CoV-2 genomes with high yield and robustness. With a shortened hands-on time from sample to virus-enriched sequencing-ready library, this rapid, versatile, and clinic-friendly approach will facilitate molecular epidemiology studies during current and future outbreaks.
分析临床样本中严重急性呼吸综合征冠状病毒 2(SARS-CoV-2)的基因组对于了解病毒的传播和进化以及疫苗的开发至关重要。现有的 RNA 测序方法对用户的技术和时间要求较高,因此不适合时间敏感的临床样本;这些方法也没有针对病毒基因组进行高性能优化。我们开发了一种简便、实用、稳健的方法,用于从临床样本中进行宏基因组和深度病毒测序。我们在从新型冠状病毒肺炎(COVID-19)患者采集的咽拭子、痰液和粪便样本中证明了我们方法的实用性,成功获得了完整的宏转录组和完整的高深度、高覆盖度 SARS-CoV-2 基因组,具有高产量和稳健性。由于从样本到病毒富集测序就绪文库的操作时间缩短,这种快速、多功能且适合临床的方法将有助于在当前和未来的爆发期间进行分子流行病学研究。