Abstract

The high variability of the influenza A virus poses a significant threat to public health, therefore monitoring viral strains and studying their genetic properties are important tasks. One part of this monitoring includes sequencing of influenza A viruses of any subtype and analysis of their whole genomes, which is especially important in cases of interspecies adaptation and reassortment. High-throughput sequencing techno-logies have significantly extended the capabilities of influenza virus epidemiological surveillance. The preparation stages for next generation sequencing (NGS) of influenza A virus include whole genome amplification using one-step RT-PCR, the results of which vary greatly depending on the sample type and quality, that, in turn, affects the coverage of virus fragments and the sequencing results in general. In this work, we propose to supplement the aforementioned technique of whole genome amplification of influenza A virus with sequential suppression PCRs to obtain an even coverage of viral segments of different lengths, which allows sequencing of samples with lower read coverage without decreasing the sequencing quality.

中文翻译：

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提高甲型流感病毒高通量测序的基因组片段覆盖率的一致性

摘要

甲型流感病毒的高变异性对公共健康构成了重大威胁，因此，监测病毒株并研究其遗传特性是重要的任务。这种监视的一部分包括对任何亚型的A型流感病毒进行测序，并对它们的整个基因组进行分析，这在种间适应和重配的情况下尤其重要。高通量测序技术已大大扩展了流感病毒流行病学监测的能力。甲型流感病毒下一代测序（NGS）的准备阶段包括使用一步式RT-PCR进行全基因组扩增，其结果根据样品类型和质量的不同而有很大差异，进而影响病毒的覆盖率片段和测序结果一般。在这项工作中