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Detection of a novel hepatitis E-like virus in faeces of wild rats using a nested broad-spectrum RT-PCR

¹ Federal Institute for Risk Assessment, Berlin, Germany;
² Institute for Hygiene and Environment Hamburg, Germany;
³ Clinic for Avian, Reptile and Fish Medicine, University of Veterinary Medicine Vienna, Austria;
⁴ Friedrich- Loeffler-Institut, Institute for Novel and Emerging Infectious Diseases, Germany

⁵ E-mail: reimar.johne{at}bfr.bund.de

Hepatitis E is a rare human disease in developed countries. It is caused by hepatitis E virus (HEV), which is most likely zoonotically transmitted to humans from domestic pigs and wild boars. Multiple reports on the detection of HEV-specific antibodies in rats suggested the presence of an HEV-related agent; however, infectious virus or a viral genome could not be demonstrated so far. Here, a nested broad-spectrum RT-PCR protocol was developed capable of detecting different HEV types including those derived from wild boar and chicken. Screening of 30 faecal samples of wild Norway rats (Rattus norvegicus) from Hamburg (Germany) resulted in the detection of two sequences with similarities to human, mammalian and avian HEV. Virus particles with morphology reminiscent of HEV were demonstrated by immune electron microscopy in one of these samples and the virus was tentatively designated as rat HEV. Genome fragments with sizes of 4,019 and 1,545 nucleotides were amplified from two samples. Sequence comparison to human and avian strains revealed only 59.9% and 49.9% sequence identity, respectively. Similarly, the deduced amino acid sequence for the complete capsid protein had 56.2% and 42.9% identity with human and avian strains, respectively. Inoculation of the samples onto three different permanent rat liver cell lines did not result in detectable virus replication as assayed by RT-PCR with cells of the fifth virus passage. Further investigations are necessary to clarify the zoonotic potential of rat HEV and to assess its suitability to serve in a laboratory rat animal model for human hepatitis E.

Received 10 September 2009; accepted 2 November 2009.