Virome Capture Sequencing Enables Sensitive Viral Diagnosis and Comprehensive Virome Analysis

摘要: Insensitivity and technical complexity have impeded the implementation of high-throughput nucleic acid sequencing in differential diagnosis viral infections clinical laboratories. Here, we describe development a virome capture platform for vertebrate viruses (VirCapSeq-VERT) that increases sensitivity sequence-based virus detection characterization. The system uses ~2 million probes cover genomes members 207 taxa known to infect vertebrates, including humans. A biotinylated oligonucleotide library was synthesized on NimbleGen cleavable array used solution-based acids present complex samples containing variable proportions host acids. use VirCapSeq-VERT resulted 100- 10,000-fold increase reads from blood tissue homogenates compared conventional Illumina using established enrichment procedures, filtration, nuclease treatments, RiboZero rRNA subtraction. had limit comparable agent-specific real-time PCR serum, blood, extracts. Furthermore, method identified novel whose were approximately 40% different designing probe library. is ideally suited analyses composition dynamics. IMPORTANCE : enables sequences sample backgrounds, those found specimens, such as tissue. highly multiplexed nature allows both simultaneous identification comprehensive genetic characterization all viruses, their variants, viruses. operational simplicity efficiency may facilitate transition diagnostic well research applications.