Enhanced Error Suppression for Accurate Detection of Low-Frequency Variants

IF 3 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS
Huimin Chen, Fei Yu, Debin Lu, Shiyue Huang, Songrui Liu, Boseng Zhang, Kunxian Shu, Dan Pu
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引用次数: 0

Abstract

The identification of low-frequency variants remains challenging due to the inevitable high error rates of next-generation sequencing (NGS). Numerous promising strategies employ unique molecular identifiers (UMIs) for error suppression. However, their efficiency depends highly on redundant sequencing and quality control, leading to tremendous read waste and cost inefficiency. Here, we describe a novel approach, enhanced error suppression strategy (EES), that addresses these challenges by (1) optimizing data utilization and reducing read waste by utilizing single-read correction that reserves abundant single reads that complement other single reads or single-strand consensus sequences (SSCSs), and (2) effectively enhancing the accuracy of NGS by employing Bayes’ theorem. EES significantly improves variant detection accuracy, achieving a background error rate of less than 4.4 × 10−5 per base pair. Additionally, the data utilization rate is dramatically increased, with a 22.9-fold enhancement in duplex consensus sequence (DCS) recovery compared to traditional methodologies. Furthermore, EES demonstrates superior error suppression performance across various base substitutions. In conclusion, EES represents a significant advancement in detecting low-frequency variants by improving data utilization and reducing sequencing errors. It potentially enhances the sensitivity and accuracy of NGS applications, proving highly valuable in clinical and research contexts where precise variant detection is critical.

增强误差抑制对低频变异的准确检测。
由于下一代测序(NGS)不可避免的高错误率,低频变异的鉴定仍然具有挑战性。许多有前途的策略使用唯一分子标识符(UMIs)来抑制错误。然而,它们的效率在很大程度上依赖于冗余的测序和质量控制,导致了巨大的读取浪费和成本低。在这里,我们描述了一种新的方法,增强错误抑制策略(EES),通过(1)优化数据利用率和减少读取浪费,利用单读校正,保留大量的单读,以补充其他单读或单链共识序列(sscs),以及(2)利用贝叶斯定理有效提高NGS的准确性来解决这些挑战。EES显著提高了变异检测精度,实现了每个碱基对的背景错误率小于4.4 × 10-5。此外,数据利用率显著提高,与传统方法相比,双工共识序列(DCS)恢复提高了22.9倍。此外,EES在各种碱基替换中表现出优越的误差抑制性能。总之,EES通过提高数据利用率和减少测序错误,在检测低频变异方面取得了重大进展。它潜在地提高了NGS应用的灵敏度和准确性,在精确变异检测至关重要的临床和研究环境中具有很高的价值。
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来源期刊
ELECTROPHORESIS
ELECTROPHORESIS 生物-分析化学
CiteScore
6.30
自引率
13.80%
发文量
244
审稿时长
1.9 months
期刊介绍: ELECTROPHORESIS is an international journal that publishes original manuscripts on all aspects of electrophoresis, and liquid phase separations (e.g., HPLC, micro- and nano-LC, UHPLC, micro- and nano-fluidics, liquid-phase micro-extractions, etc.). Topics include new or improved analytical and preparative methods, sample preparation, development of theory, and innovative applications of electrophoretic and liquid phase separations methods in the study of nucleic acids, proteins, carbohydrates natural products, pharmaceuticals, food analysis, environmental species and other compounds of importance to the life sciences. Papers in the areas of microfluidics and proteomics, which are not limited to electrophoresis-based methods, will also be accepted for publication. Contributions focused on hyphenated and omics techniques are also of interest. Proteomics is within the scope, if related to its fundamentals and new technical approaches. Proteomics applications are only considered in particular cases. Papers describing the application of standard electrophoretic methods will not be considered. Papers on nanoanalysis intended for publication in ELECTROPHORESIS should focus on one or more of the following topics: • Nanoscale electrokinetics and phenomena related to electric double layer and/or confinement in nano-sized geometry • Single cell and subcellular analysis • Nanosensors and ultrasensitive detection aspects (e.g., involving quantum dots, "nanoelectrodes" or nanospray MS) • Nanoscale/nanopore DNA sequencing (next generation sequencing) • Micro- and nanoscale sample preparation • Nanoparticles and cells analyses by dielectrophoresis • Separation-based analysis using nanoparticles, nanotubes and nanowires.
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