title: A novel pancoronavirus RT-PCR assay: frequent detection of human coronavirus NL63 in children hospitalized with respiratory tract infections in Belgium.
abstract: BACKGROUND
  Four human coronaviruses are currently known to infect the respiratory tract: human coronaviruses OC43 (HCoV-OC43) and 229E (HCoV-229E), SARS associated coronavirus (SARS-CoV) and the recently identified human coronavirus NL63 (HCoV-NL63). In this study we explored the incidence of HCoV-NL63 infection in children diagnosed with respiratory tract infections in Belgium.
  
  
  METHODS
  Samples from children hospitalized with respiratory diseases during the winter seasons of 2003 and 2004 were evaluated for the presence of HCoV-NL63 using a optimized pancoronavirus RT-PCR assay.
  
  
  RESULTS
  Seven HCoV-NL63 positive samples were identified, six were collected during January/February 2003 and one at the end of February 2004.
  
  
  CONCLUSIONS
  Our results support the notation that HCoV-NL63 can cause serious respiratory symptoms in children. Sequence analysis of the S gene showed that our isolates could be classified into two subtypes corresponding to the two prototype HCoV-NL63 sequences isolated in The Netherlands in 1988 and 2003, indicating that these two subtypes may currently be cocirculating.
authors.affiliation: Laboratory of Clinical & Epidemiological Virology, Department of Microbiology & Immunology, Rega Institute for Medical Research, University of Leuven, Belgium. elien.moes@uz.kuleuven.ac.be
authors.forename: Elien
authors.initials: E
authors.lastname: Moës
authors.forename: Leen
authors.initials: L
authors.lastname: Vijgen
authors.forename: Els
authors.initials: E
authors.lastname: Keyaerts
authors.forename: Kalina
authors.initials: K
authors.lastname: Zlateva
authors.forename: Sandra
authors.initials: S
authors.lastname: Li
authors.forename: Piet
authors.initials: P
authors.lastname: Maes
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Pyrc
authors.forename: Ben
authors.initials: B
authors.lastname: Berkhout
authors.forename: Lia
authors.initials: L
authors.lastname: van der Hoek
authors.forename: Marc
authors.initials: M
authors.lastname: Van Ranst
mesh_terms: D000293:Adolescent; D001483:Base Sequence; D001530:Belgium / Q000453:epidemiology; D002648:Child; D002675:Child, Preschool; D016384:Consensus Sequence; D017934:Coronavirus / Q000235:genetics* / Q000302:isolation & purification; D018352:Coronavirus Infections / Q000453:epidemiology / Q000821:virology*; D017931:DNA Primers / Q000737:chemistry; D005260:Female; D006801:Humans; D015994:Incidence; D007223:Infant; D008297:Male; D008969:Molecular Sequence Data; D010802:Phylogeny; D012367:RNA, Viral / Q000737:chemistry; D012141:Respiratory Tract Infections / Q000453:epidemiology / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012621:Seasons; D016415:Sequence Alignment
chemical_list: D017931:DNA Primers; D012367:RNA, Viral