AI Article Synopsis
- Ostreid herpesvirus 1 (OsHV-1) causes significant deaths in marine mollusks and is hard to study due to lack of suitable cell lines for viral replication.
- Researchers developed a method using long-range PCR to enrich OsHV-1 DNA for High-Throughput Sequencing (HTS), designing 23 primer pairs to cover almost the entire genome.
- Sequencing using both Illumina and PacBio platforms showed high genetic diversity in OsHV-1 variants and found that the virus's evolution is primarily influenced by the species of its host rather than geographical factors.
Article Abstract
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.
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| http://dx.doi.org/10.1016/j.virol.2018.09.026 | DOI Listing |
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