title: Detection of human enterovirus and human parechovirus (HPeV) genotypes from clinical stool samples: polymerase chain reaction and direct molecular typing, culture characteristics, and serotyping.
abstract: Molecular (polymerase chain reaction [PCR]) methods are increasingly used to detect and type human enteroviruses (HEVs) and parechoviruses (HPeV). Here, we assessed their value in comparison to virus culture and serotyping for detection and typing of HEV and HPeV in stool samples from hospitalized patients. By use of real-time PCR, 221/1174 patients (18.8%) were found positive for HEV/HPeV. By cell culture, a virus could be isolated from 107 of the HEV/HPeV PCR-positive samples. Culture efficiency was correlated to the Ct value, (geno)type, and cell lines used. Of the HEV/HPeV PCR-positive samples, 47% could be genotyped by VP1 genotyping and 25% by serotyping. In conclusion, PCR detection of HEV/HPeV from stool is more sensitive than virus culture, particularly for coxsackieviruses A and HPeVs. However, the genotyping method used here could identify only 47% of the HEV/HPeV strains. Further optimization and validation of direct genotyping are needed, and clinical relevance of HEV/HPeV detection in stool needs to be determined.
authors.affiliation: Department of Medical Microbiology, Laboratory of Clinical Virology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands. k.s.benschop@amc.uva.nl
authors.forename: Kimberley
authors.initials: K
authors.lastname: Benschop
authors.forename: Rene
authors.initials: R
authors.lastname: Minnaar
authors.forename: Gerrit
authors.initials: G
authors.lastname: Koen
authors.forename: Hetty
authors.initials: H
authors.lastname: van Eijk
authors.forename: Karen
authors.initials: K
authors.lastname: Dijkman
authors.forename: Brenda
authors.initials: B
authors.lastname: Westerhuis
authors.forename: Richard
authors.initials: R
authors.lastname: Molenkamp
authors.forename: Katja
authors.initials: K
authors.lastname: Wolthers
mesh_terms: D002478:Cells, Cultured; D004770:Enterovirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D004769:Enterovirus Infections / Q000175:diagnosis* / Q000821:virology; D005243:Feces / Q000821:virology*; D005838:Genotype; D006761:Hospitals; D006801:Humans; D058889:Molecular Typing; D030061:Parechovirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D010802:Phylogeny; D010850:Picornaviridae Infections / Q000175:diagnosis* / Q000821:virology; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis; D012703:Serotyping; D014776:Virus Cultivation
chemical_list: D012367:RNA, Viral