Zhou Wenxiong, Kang Li, Qiao Shuo, Duan Haifeng, Yin Chenghong, Liu Chao, Liao Zhizhao, Tang Mingchuan, Zhang Ruiying, Li Lei, Shi Lei, Du Meijie, Wang Yipeng, Yue Wentao, Xiao Yan, Di Lin, Zhang Xiannian, Pang Yuhong, Li Mingkun, Ren Lili, Wang Jianbin, Chen Zitian, Huang Yanyi
Biomedical Pioneering Innovation Center (BIOPIC), Peking-Tsinghua Center for Life Sciences, School of Life Sciences, Peking University, Beijing, China.
College of Chemistry and Molecular Engineering and Beijing National Laboratory for Molecular Sciences, Peking University, Beijing, China.
Nat Biomed Eng. 2025 Jul 2. doi: 10.1038/s41551-025-01430-8.
High-throughput sequencing technologies generate a vast number of DNA sequence reads simultaneously, which are subsequently analysed using the information contained within these fragmented reads. The assessment of sequencing technology relies on information efficiency, which measures the amount of information entropy produced per sequencing reaction cycle. Here we propose a fuzzy sequencing strategy that exhibits information efficiency more than twice that of currently prevailing cyclic reversible terminator sequencing methods. To validate our approach, we develop a fully functional and high-throughput fuzzy sequencer. This sequencer implements an efficient fluorogenic sequencing-by-synthesis chemistry and we test it across various application scenarios, including copy-number variation detection, non-invasive prenatal testing, transcriptome profiling, mutation genotyping and metagenomic profiling. Our findings demonstrate that the fuzzy sequencing strategy outperforms existing methods in terms of information efficiency and delivers accurate resequencing results with faster turnaround times.
高通量测序技术可同时生成大量DNA序列读数，随后利用这些片段化读数中包含的信息进行分析。测序技术的评估依赖于信息效率，信息效率衡量的是每个测序反应循环产生的信息熵量。在此，我们提出一种模糊测序策略，其信息效率比目前流行的循环可逆终止子测序方法高出两倍多。为验证我们的方法，我们开发了一种功能完备的高通量模糊测序仪。该测序仪实现了一种高效的荧光合成测序化学方法，我们在各种应用场景中对其进行了测试，包括拷贝数变异检测、无创产前检测、转录组分析、突变基因分型和宏基因组分析。我们的研究结果表明，模糊测序策略在信息效率方面优于现有方法，并能以更快的周转时间提供准确的重测序结果。
