Claire Crossan, Paul J. Baker, John Craft, Yasu Takeuchi, Harry R. Dalton, and Linda Scobie
Figure. . Phylogenetic analysis of HEV open reading frame 2 sequences isolated from Mytilus spp. RNA was isolated from 50–100 mg of digestive gland or gill. Tissue was homogenized in 300 μL phosphate-buffered saline, and viral RNA was isolated by using a viral RNA kit (QIAGEN, Crawley, UK), and PCR was conducted by amplifying nucleotides 6332–6476 as described (8). The nucleotide sequences were aligned and bootstrapped, and phylogenetic neighbor-joining trees were constructed by using the ClustalW software (www.ebi.ac.uk/Tools/msa/clustalw2/). Phylogenetic trees were visualized by using FigTree (http://tree.bio.ed.ac.uk/software/figtree/). Bootstrap values >70% are indicated. Scale bar indicates nucleotide substitutions per site. Sample site codes: AB, Ardrossan Beach; LB, Lunderston Bay; ABN, Aberdeen; FB, Ferrybridge; YE, Ythan Estuary. Sequences: Sw, swine; hu, human (followed by country of origin). GenBank accession numbers for reference sequences: HEV genotype 1, B73218; HEV genotype 2, M74506; HEV genotype 3, CO31008; HEV genotype 4, C272108; huUK (KernowC1), HQ389543; HuUS, JN837481; swUK AF503512; huFrance, JN906974; swCanada, AY115488; swSpain, JQ522948; sw2Japan AB248521, huJapan AB161719.