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

被引:307
作者
Johne, Reimar [1 ]
Plenge-Boenig, Anita [2 ]
Hess, Michael [3 ]
Ulrich, Rainer G. [4 ]
Reetz, Jochen [1 ]
Schielke, Anika [1 ]
机构
[1] Fed Inst Risk Assessment, D-12277 Berlin, Germany
[2] Inst Hyg & Environm Hamburg, D-20539 Hamburg, Germany
[3] Univ Vet Med, Clin Avian Reptile & Fish Med, A-1210 Vienna, Austria
[4] Inst Novel & Emerging Infect Dis, Friedrich Loeffler Inst, D-17493 Greifswald, Germany
关键词
CELL-CULTURE-SYSTEM; UNITED-STATES; BLOOD-DONORS; IN-VIVO; EXPERIMENTAL-INFECTION; PHYLOGENETIC ANALYSIS; WIDESPREAD INFECTION; PREVALENCE; SWINE; ANTIBODIES;
D O I
10.1099/vir.0.016584-0
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Hepatitis E is a rare human disease in developed countries. It is caused by hepatitis E virus (HEV), which is probably transmitted zoonotically to humans from domestic pigs and wild boars. Multiple reports on the detection of HEV-specific antibodies in rats have suggested the presence of an HEV-related agent; however, infectious virus or a viral genome has not been 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 from 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 a morphology reminiscent of HEV were demonstrated by immunoelectron microscopy in one of these samples and the virus was tentatively designated rat HEV. Genome fragments with sizes of 4019 and 1545 nt were amplified from two samples. Sequence comparison with 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.
引用
收藏
页码:750 / 758
页数:9
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