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. 2009 Oct;46(2):140-4.
doi: 10.1016/j.jcv.2009.07.007. Epub 2009 Aug 3.

Genotypic characterization of symptomatic hepatitis E virus (HEV) infections in Egypt

Jason T Blackard  1 , Susan D Rouster, Soad Nady, Gehan Galal, Naglaa Marzuuk, Marwaa M Rafaat, Enas Daef, Salwa Seif El Din, Robert H Purcell, Suzanne U Emerson, Kenneth E Sherman, M Tarek Shata
Affiliations

Genotypic characterization of symptomatic hepatitis E virus (HEV) infections in Egypt

Jason T Blackard et al. J Clin Virol. 2009 Oct.

Abstract

Background: Hepatitis E virus (HEV) is a common cause of acute viral hepatitis (AVH) in many developing countries. In Egypt, HEV seroprevalence is among the highest in the world; however, only a very limited number of Egyptian HEV sequences are currently available.

Objectives: The objectives were to determine the HEV genotype(s) currently circulating in Egypt.

Study design: AVH patients without serologic evidence of hepatitis A, B, and C viruses were evaluated for possible HEV infection using serologic assays for anti-HEV IgM and anti-HEV IgG and real-time PCR for HEV RNA. Stool suspensions from suspected cases were inoculated into rhesus macaques to confirm the presence of HEV. Sequence analysis was utilized to determine HEV genotype.

Results: Of 287 subjects with AVH enrolled, 58 had serologic evidence of acute HEV infection. Stool samples for two of these patients were repeatedly positive for HEV RNA by real-time PCR. Macaques experimentally inoculated with these human stools also developed viremia. Sequence analysis of open reading frame (ORF) 1 demonstrated that these isolates belonged to HEV genotype 1 and were 3.9-9.5% divergent from other genotype 1 isolates. ORF2 was 5.3-8.7% divergent from previously reported Egyptian isolates.

Conclusions: This study strongly suggests that genotype 1 HEV related to other North African isolates is circulating in acute symptomatic patients in Egypt. Further evaluation of genotypic variability is underway in this highly endemic cohort and is considered an important component of our increased understanding of HEV pathogenesis.

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Conflict of interest statement

Competing interests: None declared

Figures

Figure 1
Figure 1
Phylogenetic analysis of HEV sequences from two symptomatic Egyptian patients. A) ORF 1 (242 base pairs [bp]; B) ORF2/3 overlap region (97 bp); C) ORF 2 (98 bp). Reference sequences are denoted by their GenBank accession numbers and the following country-specific abbreviations: Mexico (MX), United States (US), Japan (JP), China (CH), Chad (CD), Morocco (MO), India (IN), Pakistan (PA), Nepal (NP), Egypt (EG), and Algeria (AL). The two Egyptian samples characterized in the current study are highlighted in bold. The statistical robustness and reliability of the branching order within each phylogenetic tree were confirmed by bootstrap analysis using 100 replicates . Sequences from the current study – with primer sequences removed – have been submitted to GenBank under accession numbers FJ423076 – FJ423080.
Figure 1
Figure 1
Phylogenetic analysis of HEV sequences from two symptomatic Egyptian patients. A) ORF 1 (242 base pairs [bp]; B) ORF2/3 overlap region (97 bp); C) ORF 2 (98 bp). Reference sequences are denoted by their GenBank accession numbers and the following country-specific abbreviations: Mexico (MX), United States (US), Japan (JP), China (CH), Chad (CD), Morocco (MO), India (IN), Pakistan (PA), Nepal (NP), Egypt (EG), and Algeria (AL). The two Egyptian samples characterized in the current study are highlighted in bold. The statistical robustness and reliability of the branching order within each phylogenetic tree were confirmed by bootstrap analysis using 100 replicates . Sequences from the current study – with primer sequences removed – have been submitted to GenBank under accession numbers FJ423076 – FJ423080.
Figure 1
Figure 1
Phylogenetic analysis of HEV sequences from two symptomatic Egyptian patients. A) ORF 1 (242 base pairs [bp]; B) ORF2/3 overlap region (97 bp); C) ORF 2 (98 bp). Reference sequences are denoted by their GenBank accession numbers and the following country-specific abbreviations: Mexico (MX), United States (US), Japan (JP), China (CH), Chad (CD), Morocco (MO), India (IN), Pakistan (PA), Nepal (NP), Egypt (EG), and Algeria (AL). The two Egyptian samples characterized in the current study are highlighted in bold. The statistical robustness and reliability of the branching order within each phylogenetic tree were confirmed by bootstrap analysis using 100 replicates . Sequences from the current study – with primer sequences removed – have been submitted to GenBank under accession numbers FJ423076 – FJ423080.
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