African Swine Fever Virus (ASFV) causes a transmissible and fatal disease in pigs that is currently devastating global swine production. Efficient and economical collection of genetic data from ASFV field isolates is essential for bio-surveillance, to limit and control its spread, and to better understand ASF disease ecology. Standard genotyping and subtyping of ASFV field isolates is currently limited to a few variable regions within the ASFV genome. However, more extensive sequencing is necessary to better understand ASFV molecular evolution and identify regions relevant to genetic diversity. In this study, we developed a method for rapid and efficient next generation sequencing of approximately 40% of the ASFV genome using long PCR amplification of six different genomic regions. The amplified regions contain all segments currently used for genotyping and additional genes predicted to contribute to ASFV diversity. The primers used for amplification are broadly compatible with published ASFV genomes, permitting their use on relevant ASFV isolates. This methodology provides the enhanced depth of coverage of amplicon-based sequencing while mitigating complications associated with ASFV whole-genome sequencing. Implementation of this methodology could substantially increase the scale of ASFV genetic data collection, which is necessary to effectively monitor and combat this critical agricultural disease.

非洲猪瘟病毒（ASFV）引起了猪的一种传播性和致命性疾病，目前正在破坏全球的猪生产。从ASFV野外分离株中高效而经济地收集遗传数据对于生物监测，限制和控制其传播以及更好地了解ASF疾病生态至关重要。ASFV田间分离株的标准基因分型和亚型目前仅限于ASFV基因组内的几个可变区。但是，需要更广泛的测序以更好地了解ASFV分子进化并鉴定与遗传多样性相关的区域。在这项研究中，我们开发了一种方法，可使用六个不同基因组区域的长PCR扩增，对约40％的ASFV基因组进行快速高效的下一代测序。扩增的区域包含当前用于基因分型的所有片段以及预计有助于ASFV多样性的其他基因。用于扩增的引物与已发表的ASFV基因组广泛兼容，从而可以在相关的ASFV分离株上使用。这种方法可提高基于扩增子的测序的覆盖深度，同时减轻与ASFV全基因组测序相关的并发症。这种方法的实施可能会大大增加ASFV遗传数据收集的规模，这对于有效监测和对抗这种关键的农业疾病是必要的。这种方法可提高基于扩增子的测序的覆盖深度，同时减轻与ASFV全基因组测序相关的并发症。这种方法的实施可能会大大增加ASFV遗传数据收集的规模，这对于有效监测和对抗这种关键的农业疾病是必要的。这种方法可提高基于扩增子的测序的覆盖深度，同时减轻与ASFV全基因组测序相关的并发症。这种方法的实施可能会大大增加ASFV遗传数据收集的规模，这对于有效监测和对抗这种关键的农业疾病是必要的。