Approcci innovativi per la diagnosi e caratterizzazione di virus emergenti

The evolution of approaches for viral identification and characterization has significantly prompted the research in virology in recent years. Classical approaches provided fundamental knowledge but displayed limitations in exploring viromes. Metagenomics techniques such as sequence independent single primer amplification (SISPA), random amplification, rolling circle amplification (RCA), and degenerate oligonucleotide primed PCR (DOP-PCR), have demonstrated able to amplify viral genomes with many advantages and few drawbacks. Sequencing approaches of second (i.e., Illumina, Ion Torrent) and third (Oxford Nanopore Sequencing, PacBio) generation, associated with metagenomics, have enabled mass production of data on novel viruses, posing new challenges in deciphering metagenome-assembled viruses and their effects on hosts. The aim of the present thesis is to report the diagnosis and characterization of viruses, using a multitude of protocols and procedures, newly developed or edited from the literature, taking advantage of novel sequencing technologies and bioinformatic tools. These novel approaches are more often integrated in the research and whenever possible in the diagnostic routine of the virology laboratory.