Detection and Epidemiological Analysis of Symbiotic Viruses from Protozoa Using the FLDS (A Comprehensive dsRNA Sequencing Method)

概要

We have detected the symbiotic virus of Eimeria that infects chickens by next-generation sequencing (NGS) analysis using the FLDS method (Fragmented and primer Ligated dsRNA Sequencing). Total nucleic acid was extracted from Eimeria oocysts and dsRNA was purified using a cellulose column that specifically adsorbed dsRNA. Subsequent analysis with NGS using the FLDS method yielded various dsRNA contigs. Of these, the percentage of total leads indicates that there is a high probability that three types of contigs are present in chicken Eimeria or chickens. As a result of BLAST analysis, 1 contig showed more than 80% homology to Eimeria brunetti RNA virus 1, and this contig was considered to be the sequence of the symbiotic virus of Eimeria. The remaining two species were suggested to be novel dsRNA viruses.

Next, we designed primers using the analyzed sequences and carried out epidemiological analysis of Eimeria brunetti RNA virus 1 by PCR. As a result, viral sequences were detected from some chicken Eimeria in Japan.