Detection and characterization of human parechoviruses in archived cell cultures, in Hungary

Abstract

Background Human parechoviruses (HPeV) belonging to the family Picornaviridae are widespread enteric pathogens and associated with various clinical symptoms in humans. Objectives There is no report for detection of the circulating parechoviruses in Central Europe. The aim of this retrospective study was to detect and characterize human parechoviruses in cell cultures with “enterovirus”-like cytophatic effect (CPE) archived between 1990 and 2000, in Hungary. Study design Fecal samples from children with symptoms of gastroenteritis under age of 10 were cultured as a previous routine diagnostic laboratory protocol for “enterovirus”. Cell cultures indicating CPE were archived and deeply chilled (−80 °C) from minimum 2 individuals (2–12 patients) in each year. Specimens were tested retrospectively, in 2009, for HPeV by reverse transcription-PCR (RT-PCR) method using 5′UTR conserved primers. Specific primer pairs were designed to determine the complete nucleotide sequence of the structural region (VP0–VP3–VP1) of HPeV. Results Nine (9.1%) of 66 archived samples were found to be HPeV-positive. Six (67%) samples were identified as HPeV1, 2 (22%) were HPeV4 and 1 could not be determined. Three HPeV1 clusters were identified according to the isolation date originated from years 1990/1991, 1992/1995 and 1998. Conclusions This is the first detection of human parechoviruses in Central Europe. Detection and genetic characterization of HPeV in available historical samples infected with previously unidentifiable agents with “enterovirus-like” cytopathogenic effect help to understand the genetic diversity and evolution of these viruses.