纳米孔测序数据比对分析方法与参考数据库研究进展

Research progress on nanopore sequencing data alignment analysis methods and reference databases

摘要:

纳米孔测序作为测序技术的新兴热点，凭借其读长较长、检测快速和设备紧凑小巧等独特优势，在物种鉴定、基因组组装、变异检测、转录组分析等领域展现出巨大潜力。然而，纳米孔测序数据错误率较高，存在序列插入和缺失等问题，对传统序列比对工具运用和参考数据库构建提出了新的挑战。本文围绕纳米孔数据特征，系统梳理了适配纳米孔测序的序列比对工具，针对长读长测序、实时测序、错误率兼容、宏基因组和结构变异检测这5种不同应用场景，阐述了其在处理序列数据时的优势及局限性；同时，还从数据源的角度对序列参考基因组数据库进行多维度分类整理，并总结了纳米孔高质量数据库构建的关键技术。本文通过对比对工具与数据库进行协同分析，为纳米孔测序数据分析的优化与创新提供参考，推动宏基因组测序从数据生成向功能解析的深度转化。

Abstract:

Nanopore sequencing, as an emerging hotspot in sequencing technology, demonstrates tremendous potential in species identification, genome assembly, variant detection, and transcriptome analysis, owing to its distinctive advantages including extended read lengths, rapid detection capabilities, and compact instrumentation. However, nanopore sequencing data are characterized by high error rates and presence of insertions and deletions, which pose novel challenges for the application of conventional sequence alignment tools and the construction of reference databases. Focusing on the characteristics of nanopore data, this paper systematically sorts out sequence alignment tools suitable for nanopore sequencing, and elaborates on their advantages and limitations in processing sequence data for five different application scenarios: long-read sequencing, real-time sequencing, error rate compatibility, metagenomics, and structural variation detection. Meanwhile, from the perspective of data sources, this paper conducts multi-dimensional classification and organization of reference genome databases, and sorts out the key technologies for constructing high-quality nanopore databases. Through the collaborative analysis of alignment tools and databases, this paper provides references for the optimization and innovation of nanopore sequencing data analysis, and promotes the in-depth transformation of metagenomic sequencing from data generation to functional analysis.