Detection and characterization of human parechoviruses in archived cell cultures, in Hungary.
BACKGROUND
Human parechoviruses (HPeV) belonging to the family Picornaviridae are widespread enteric pathogens and associated with various clinical symptoms in humans.


OBJECTIVE
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.


METHODS
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 degrees 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.
Regional Laboratory of Virology, National Reference Laboratory of Gastroenteric Viruses, ANTSZ Regional Institute of State Public Health Service, H-7623 Szabadság út 7, Pécs, Hungary.
Akos
A
Boros
Mária
M
Uj
Péter
P
Pankovics
Gábor
G
Reuter
D018929:Cell Culture Techniques; D002648:Child; D002675:Child, Preschool; D016000:Cluster Analysis; D003588:Cytopathogenic Effect, Viral; D017931:DNA Primers / Q000235:genetics; D005243:Feces / Q000821:virology; D006801:Humans; D006814:Hungary; D007223:Infant; D008969:Molecular Sequence Data; D030061:Parechovirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D010802:Phylogeny; D010850:Picornaviridae Infections / Q000175:diagnosis*; D012189:Retrospective Studies; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D014764:Viral Proteins / Q000235:genetics
D017931:DNA Primers; D014764:Viral Proteins