Conversion Of Viral RNA Research Articles

The Oxford Nanopore MinION as a Versatile Technology for the Diagnosis and Characterization of Emerging Plant Viruses.

The emergence of novel viral epidemics that could affect major crops represents a serious threat to global food security. The early and accurate identification of the causative viral agent is the most important step for a rapid and effective response to disease outbreaks. Over the last years, the Oxford Nanopore Technologies (ONT) MinION sequencer has been proposed as an effective diagnostic tool for the early detection and identification of emerging viruses in plants, providing many advantages compared with different high-throughput sequencing (HTS) technologies. Here, we provide a step-by-step protocol that we optimized to obtain the virome of "Lamon bean" plants (Phaseolus vulgaris L.), an agricultural product with Protected Geographical Indication (PGI) in North-East of Italy, which is frequently subjected to multiple infections caused by different RNA viruses. The conversion of viral RNA in ds-cDNA enabled the use of Genomic DNA Ligation Sequencing Kit and Native Barcoding DNA Kit, which have been originally developed for DNA sequencing. This allowed the simultaneous diagnosis of both DNA- and RNA-based pathogens, providing a more versatile alternative to the use of direct RNA and/or direct cDNA sequencing kits.

Interaction of viral RNA with Escherichia coli: III. Synthesis of poliovirus-specific RNA directed by isolated poliovirus RNA

Poliovirus RNA directs the synthesis of virus-specific RNA in E. coli as reported previously for poliovirus-induced double-stranded RNA. Synthesis of viral RNA can be followed by conversion of viral RNA into a double-stranded RNase-resistant state, by increase in infectivity and by hybridization of newly synthesized RNA to viral RNA. Virus-specific RNA synthesis occurs also in the presence of inhibitors of protein synthesis indicating that an enzyme is present in E. coli which can use RNA as a template.