Abstract

MONTALVO VILLALBA, María de la Caridad et al. Isolation and propagation of hepatitis E virus in several cell lines. Rev Cubana Med Trop [online]. 2008, vol.60, n.3. ISSN 0375-0760.

BACKGROUND: hepatitis E virus (HEV) is the causative agent of hepatitis E. Biological and molecular properties of Asian HEV strains have been explored in cells cultures. OBJECTIVES: the aim of this investigation was to isolate and propagate a Cuban HEV isolate in different cell lines. METHODS: A549 cells monolayer was infected with faeces suspension from patient with sporadic hepatitis E and followed up until tenth passage. Lately, the supernatant harvested from third passage in A549 cells was inoculated in MRC5, HEP-2, LLCMK2, HeLa y FRhK4 for propagation study. These cells were observed up to third passage. RNA and viral antigen of ECV/2349-03 HEV strain were identified by RT-PCR and indirect immunofluorescence. RESULTS: CPE appeared since first passage, at third day of post-inoculation in A549 cells. HEV antigens and genome were detected in all serial passages. CPE was not observed in the rest of cellular cultures. In the cells used for propagation the viral genome was observed from first passage, while the antigens were detected since second passage, except HeLa. CONCLUSIONS: these results confirm that A549 can be used to isolate and propagate HEV. Meanwhile, the MRC5, HEP-2, LLCMK2 and FRhK4 were able to support viral growing.

Keywords : HEV; A549 cells; propagation.