Foreign Animal Disease Diagnostic Laboratory, National Veterinary Services Laboratories, Animal and Plant Health Inspection Service, U.S. Department of Agriculture, Plum Island Animal Disease Center, Orient, NY 11944, USA.
Agricultural Research Service, U.S. Department of Agriculture, Plum Island Animal Disease Center, Orient, NY 11944, USA.
Viruses. 2024 Sep 26;16(10):1522. doi: 10.3390/v16101522.
African swine fever (ASF) is an acute, highly hemorrhagic viral disease in domestic pigs and wild boars. The disease is caused by African swine fever virus, a double stranded DNA virus of the family. ASF can be classified into 25 different genotypes, based on a 478 bp fragment corresponding to the C-terminal sequence of the B646L gene, which is highly conserved among strains and encodes the major capsid protein p72. The C-terminal end of p72 has been used as a PCR target for quick diagnosis of ASF, and its characterization remains the first approach for epidemiological tracking and identification of the origin of ASF in outbreak investigations. Recently, a new classification of ASF, based on the complete sequence of p72, reduced the 25 genotypes into only six genotypes; therefore, it is necessary to have the capability to sequence the full-length B646L gene (p72) in a rapid manner for quick genotype characterization. Here, we evaluate the use of an amplicon approach targeting the whole B646L gene, coupled with nanopore sequencing in a multiplex format using Flongle flow cells, as an easy, low cost, and rapid method for the characterization and genotyping of ASF in real-time.
非洲猪瘟（ASF）是一种急性、高出血性的猪和野猪病毒病。该病由非洲猪瘟病毒引起，这是一种双链 DNA 病毒，属于家族。根据 B646L 基因 C 末端序列的 478bp 片段，ASF 可分为 25 种不同的基因型，该片段在株间高度保守，编码主要衣壳蛋白 p72。p72 的 C 末端已被用作 ASF 快速诊断的 PCR 靶标，其特征仍然是流行病学追踪和暴发调查中 ASF 起源鉴定的首要方法。最近，基于 p72 的完整序列对 ASF 进行了新的分类，将 25 种基因型减少到仅 6 种基因型；因此，有必要快速地对全长 B646L 基因（p72）进行测序，以便快速进行基因型特征描述。在此，我们评估了一种扩增子方法的应用，该方法靶向整个 B646L 基因，并结合 Flongle 流控芯片的多重纳孔测序，作为一种简单、低成本、快速的实时 ASF 特征描述和基因分型方法。
