Introduction: Enteric viruses, including Hepatitis E virus (HEV), are able to persist under environmental conditions and may cause public health problems by contaminating natural and drinking water resources. Routine procedures for monitoring viruses in water samples have not been established for the water microbiology screening panel. Methodology: Eighty-six raw sewerage samples were collected from the different regions of Islamabad and Rawalpindi, the twin cities of Pakistan. Samples were concentrated for HEV, using a polyethylene glycol-based method followed by viral RNA extraction using a commercial kit-based method. Reverse transcription polymerase chain reaction (RT-PCR) with HEV specific primers was used for the detection of HEV.Results: The present investigation focused on 86 raw sewerage water samples taken from different locations of drainage outlets of Islamabad and Rawalpindi. After careful experimentation, 35 samples were found to be RT-PCR positive. Nineteen (44.7%) out of 47 samples from Rawalpindi city were HEV positive while 16 (41.02%) out of 39 samples from Islamabad were HEV positive. All positive samples were found in the highly congested areas. Conclusions: The high detection rate of HEV in this study shows that HEV circulates at a relatively high frequency in the sewerage waters in Pakistan. This study is the first report on detection of HEV from sewerage waste water from Pakistan and suggests that HEV might be a potent indicator of viral pollution in environmental specimens.

Background: Vaccine development against emerging infections is essentially important for saving people from increasing viral infections. In developing countries, Hepatitis E (HEV) is a common infection affecting millions of people worldwide. Based on In-silico analysis, different approaches have been targeted. Objectives: Rationale of this study is to design an epitope-based vaccine candidates with the help of immunoinformatics that can predict promiscuous B-cell and T-cell epitopes of the most antigenic HEV-ORF2 capsid protein. Materials & Methods: This study suggests potential T-cell and B-cell epitopes of the highly antigenic HEV ORF2 capsid protein while using various In-silico tools such as NCBI-BLAST, Expassy, CLC workbench, Ellipro and Discotope. Results: Potential antigenic and immunogenic CD8+ T-cell epitopes were predicted from the global consensus sequence of ORF2-HEV. Furthermore, twenty-two linear B-cell epitopes were predicted. Among these, “SLGAGPV” at position 587-593 and “LEFRNLTPGNTNTRVSRYSS” at position 306-325 were most antigenic with antigenicity score 1.4206 and 1.3600 respectively. Discontinuous B-cell epitopes were found by three-dimensional capsid protein structure. Epitopes predicted in this study reveal high antigenicity and promiscuity for HLA classes. Conclusion: Collectively, our data suggests promiscuous epitopes that can potentially acts as new candidates for the design of HEV peptide vaccine. Keywords: Virology; gastrointestinal disease.