混合文库测序数据量优化提升肿瘤外科基因检测效能的临床应用与验证

doi:10.3969/j.issn.1009-976X.2026.01.004

岭南现代临床外科 ›› 2026, Vol. 26 ›› Issue (01): 27-32.DOI: 10.3969/j.issn.1009-976X.2026.01.004

混合文库测序数据量优化提升肿瘤外科基因检测效能的临床应用与验证

骆嘉欢¹, 黄靖华¹, 蒋圆玲¹, 黄永胜^1,2, 尹欣科¹, 付莎¹, 欧阳能太¹, 廖健伟^1,2,*

1.中山大学孙逸仙纪念医院细胞分子诊断中心,广东广州 510120;
2.广东省恶性肿瘤表现遗传与基因调控重点实验室,广州 510120

通讯作者: *廖健伟,Email：liaojw8@mail.sysu.edu.cn
基金资助:
广东省基础与应用基础研究基金（2021A1515111138）; 广州市科技计划项目（2023A04J2103）

Data volume optimization of pooled library sequencing enhances clinical application and validation of genetic testing efficacy in oncological surgery

LUO Jiahuan¹, HUANG Jinghua¹, JIANG Yuanling¹, HUANG Yongsheng^{1, 2}, YIN Xinke¹, FU Sha¹, OUYANG Nengtai¹, LIAO Jian-wei^1,2,*

1. Cellular & Molecular Diagnostics Center, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, China;
2. Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Guangzhou 510120, China

Received:2025-12-06 Online:2026-02-20 Published:2026-03-24
Contact: *LIAO Jianwei, liaojw8@mail.sysu.edu.cn

摘要/Abstract

摘要： 目的优化不同长度肿瘤文库在Illumina NextSeq 550测序平台上的数据分配策略,在保障检测质量的前提下满足外科诊疗的时效性要求。方法通过10个批次实验,量化3种长度文库（扩增子275 bp/300 bp,杂交捕获350 bp）的产出比（产出数据量/上机数据量）,得出不同文库的成簇能力,进而分析得出不同长度文库数据量配比的调整区间,以此调整各文库的浓度以调控其最终数据量。结果 300 bp扩增子文库数据量需下调至0.55~0.72倍, 275 bp扩增子文库数据量需下调至0.56~0.79倍,350 bp杂交捕获文库数据量按芯片数据冗余量上调后,可平衡数据产出。结论基于产出比量化不同长度文库的簇生成能力差异来调整文库上机数据量,可实现肿瘤混合测序数据均衡产出及芯片数据的合理利用。

关键词: 肿瘤靶向测序, 数据量分配, NextSeq 550测序平台, 杂交捕获文库, 扩增子文库

Abstract: Objective To optimize data allocation strategies for tumor libraries of different lengths on the Illumina NextSeq 550 sequencing platform, ensuring detection quality while meeting the timeliness requirements of surgical diagnosis and treatment. Methods In 10 batches of experiments, the output ratio （output data volume/input data volume） of three library types （275 bp/300 bp amplicons, 350 bp hybrid capture） was quantified.This quantification was used to determine the cluster generation capability of different libraries. Further analysis was conducted to derive the adjustment range for the data volume ratio of libraries with different lengths. Based on this range, the concentration of each library was adjusted to regulate its final data volume. Results The data volume for 300 bp amplicon libraries should be adjusted to a range of 0.55× to 0.72×. The data volume of 275 bp amplicon libraries should be adjusted in the range of 0.56× to 0.79×. The data volume of 350 bp hybridization capture libraries can be adjusted according to the amount of redundant data on the flow cell to balance data output. Conclusion Adjusting library loading volumes based on output ratios quantifying clustering capabilities across libraries of varying lengths enables balanced output of tumor pooled sequencing data and optimal utilization of chip resources.

Key words: tumor target sequencing, data assignment, NextSeq 550 sequencing platform, hybridization capture-based enrichment, amplicon enrichment

中图分类号:

R730.4
R446

骆嘉欢, 黄靖华, 蒋圆玲, 黄永胜, 尹欣科, 付莎, 欧阳能太, 廖健伟. 混合文库测序数据量优化提升肿瘤外科基因检测效能的临床应用与验证[J]. 岭南现代临床外科, 2026, 26(01): 27-32.

LUO Jiahuan, HUANG Jinghua, JIANG Yuanling, HUANG Yongsheng, YIN Xinke, FU Sha, OUYANG Nengtai, LIAO Jian-wei. Data volume optimization of pooled library sequencing enhances clinical application and validation of genetic testing efficacy in oncological surgery[J]. Lingnan Modern Clinics In Surgery, 2026, 26(01): 27-32.

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混合文库测序数据量优化提升肿瘤外科基因检测效能的临床应用与验证

Data volume optimization of pooled library sequencing enhances clinical application and validation of genetic testing efficacy in oncological surgery

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