Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology; Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, Shandong, China.
Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER), Laboratoire de Génétique et Pathologie (LGP), Avenue de Mus de Loup, 17390 La Tremblade, France.
Virology. 2019 Jan 2;526:81-90. doi: 10.1016/j.virol.2018.09.026. Epub 2018 Oct 24.
Ostreid herpesvirus 1 (OsHV-1) is an important pathogen associated with mass mortalities of cultivated marine mollusks worldwide. Since no cell line allows OsHV-1 replication in vitro, it is difficult to isolate enough high-purity viral DNA for High-Throughput Sequencing (HTS). We developed an efficient approach for the enrichment of OsHV-1 DNA for HTS with long-range PCR. Twenty-three primer pairs were designed to cover 99.3% of the reference genome, and their performances were examined on ten OsHV-1 infected samples. Amplicon mixtures from six successfully amplified samples were sequenced with Illumina platform, and one of them (ZK0118) was also sequenced with the PacBio platform. PacBio reads were assembled into 2 scaffolds compared to 9-68 scaffolds assembled from the Illumina reads. Genomic comparison confirmed high genetic diversity among OsHV-1 variants. Phylogenetic analysis revealed that OsHV-1 evolution was mainly impacted by its host species rather than spatial segregation.
牡蛎疱疹病毒 1（OsHV-1）是一种与全球养殖海洋贝类大量死亡相关的重要病原体。由于没有细胞系可以在体外复制 OsHV-1，因此很难分离出足够高纯度的病毒 DNA 进行高通量测序（HTS）。我们开发了一种利用长距离 PCR 高效富集 OsHV-1 DNA 进行 HTS 的方法。设计了 23 对引物来覆盖参考基因组的 99.3%，并在 10 个 OsHV-1 感染样本中检查了它们的性能。从 6 个成功扩增的样本中扩增的混合物进行了 Illumina 平台测序，其中一个（ZK0118）也进行了 PacBio 平台测序。PacBio 读取被组装成 2 个支架，而 Illumina 读取组装成 9-68 个支架。基因组比较证实了 OsHV-1 变体之间存在高度遗传多样性。系统发育分析表明，OsHV-1 的进化主要受其宿主物种的影响，而不是空间隔离。
