Next-generation sequencing (NGS) has revolutionized the scale and depth of biomedical sciences. Because of its unique ability for the detection of sub-clonal variants within genetically diverse populations, NGS has been successfully applied to analyze and quantify the exceptionally-high diversity within viral quasispecies, and many low-frequency drug- or vaccine-resistant mutations of therapeutic importance have been discovered. Although many works have intensively discussed the latest NGS approaches and applications in general, none of them has focused on applying NGS in viral quasispecies studies, mostly due to the limited ability of current NGS technologies to accurately detect and quantify rare viral variants. Here, we summarize several error-correction strategies that have been developed to enhance the detection accuracy of minority variants. We also discuss critical considerations for preparing a sequencing library from viral RNAs and for analyzing NGS data to unravel the mutational landscape.

中文翻译：

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应用下一代测序来解开病毒准种的突变格局。

新一代测序 (NGS) 彻底改变了生物医学科学的规模和深度。由于其在基因多样性人群中检测亚克隆变异的独特能力，NGS 已成功应用于分析和量化病毒准种内异常高的多样性，以及许多低频药物或疫苗抗性突变。重要性已被发现。尽管许多工作对最新的 NGS 方法和应用进行了深入的讨论，但没有一个专注于将 NGS 应用于病毒准种研究，这主要是由于当前 NGS 技术准确检测和量化稀有病毒变体的能力有限。在这里，我们总结了几种已开发的纠错策略，以提高少数变体的检测精度。