Department of Biochemistry and Immunology, National Institute of Veterinary Research, Hanoi, Vietnam.
Methods Mol Biol. 2022;2503:205-215. doi: 10.1007/978-1-0716-2333-6_16.
Next-generation sequencing (NGS) technologies have been powerfully applied in both research and clinical settings for the understanding and control of infectious disease. It enables high-resolution characterization of viral pathogens in terms of properties that include molecular epidemiology, genotype, serotype, and virulence. However, a beginner's NGS protocol for characterization of African swine fever virus (ASFV) is lacking. Here, we present detailed step-by-step methods for obtaining NGS data from ASF virus (ASFV) using the Illumina platform. The protocol has been performed with respect to ASFV DNA genome extraction, qualification of DNA, library preparation, quality control, de novo assembly, and data quality control. The protocol represents a step-by-step and reproducible method for producing high-quality sequencing data. The key advantages of this protocol include the protocol being very simple for users with no experience of genome sequencing and reproducibility of the protocol for other DNA genome viruses.
下一代测序（NGS）技术在研究和临床环境中都得到了广泛应用，可用于了解和控制传染病。它能够从分子流行病学、基因型、血清型和毒力等方面对病毒病原体进行高分辨率的描述。然而，目前缺乏用于描述非洲猪瘟病毒（ASFV）的新手 NGS 方案。在这里，我们使用 Illumina 平台展示了从 ASF 病毒（ASFV）获取 NGS 数据的详细分步方法。该方案已经针对 ASFV DNA 基因组提取、DNA 质量评估、文库制备、质量控制、从头组装和数据质量控制进行了操作。该方案代表了一种用于生成高质量测序数据的逐步和可重复的方法。该方案的主要优点包括该方案对没有基因组测序经验的用户来说非常简单，并且该方案对于其他 DNA 基因组病毒具有可重复性。
