Invasive Species Compendium

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Abstract

Novel hepatitis E virus in ferrets, the Netherlands.

Abstract

A study was conducted to investigate hepatitis E virus (HEV) sequences in ferrets (Mustela putorius) from the Netherlands, using random polymerase chain reaction (PCR) amplification and high-throughput sequencing technology. Faecal samples were collected from ferrets in 2010. Viral RNA was isolated and viral metagenomic libraries were constructed for 454 pyrosequencing and 248 840 sequence reads were generated from 7 faecal samples. Using Blastn and Blastx, 289 sequence reads were identified in one sample that were related to rat HEV and that could be assembled into 6 contigs covering ∼50% of the ferret HEV (FRHEV) genome. A set of nested PCR primers was developed based on obtained sequences to detect viral RNA. Total RNA extracted from 43 household pet ferret faecal samples collected from 19 locations was used to perform reverse transcription PCR amplification. Using this PCR, viral RNA was detected in 4 (9.3%) faecal samples tested from 4 locations (distance between each sampling location ranged from 25 to 127 km). All amplicons were confirmed by nucleotide sequencing. Two PCR-positive samples (FRHEV4 and FRHEV20) were selected to characterize the complete genome of this virus. Using overlapping fragments, 2 complete FRHEV genome sequences were assembled that contained 6854 nt, including a 13-nt 3′ poly A tail and a 12-nt 5′ end. The full-genome sequences of FRHEV4 and FRHEV20 showed 98.6% sequence identity and were deposited into GenBank database under accession numbers JN998606 and JN998607, respectively. Phylogenetic analysis of the complete genomes showed that FRHEV was separated from genotype 1-4 HEVs and clustered with rat HEV. It is suggested that further studies are needed to identify the zoonotic potential of FRHEV.