title: Detection and characterisation of human metapneumovirus from children with acute respiratory symptoms in north-west England, UK.
abstract: BACKGROUND
  Human metapneumovirus (hMPV) causes a spectrum of respiratory disease ranging from trivial coryzal symptoms to fatal pneumonia, with a predilection for the very young, the immune suppressed and the frail elderly. Five distinct lineages of the virus genome have been described.
  
  
  OBJECTIVE
  To develop and evaluate a sensitive, real-time PCR (RT-PCR) assay capable of detecting all lineages of hMPV, suitable for use in a diagnostic laboratory.
  
  
  METHODS
  An RT-PCR assay was developed using novel primers and dual-labelled minor-groove-binding (MGB) probes complementary to consensus sequences. The assay and two alternative assays were tested against external quality assurance (EQA) panels. 221 respiratory samples collected during 2003-2004 were screened using the new assay. hMPV positive samples were sequenced and phylogenetically analysed.
  
  
  RESULTS
  Three genetic lineages of hMPV were detected during 2003-2004. Incidence was low (2.3%) compared to previous years. All five lineages had been present in the same community within the past 3 years.
  
  
  CONCLUSIONS
  The new assay correctly identified more EQA samples, including those at greatest dilution, than the alternative assays and detected all five lineages. Seasonal circulation of hMPV in paediatric patients with acute respiratory symptoms is dynamic with respect to incidence and viral genotype.
authors.affiliation: Liverpool Specialist Virology Centre, Royal Liverpool University Hospital, Liverpool, UK.
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Hopkins
authors.forename: Catherine
authors.initials: C
authors.lastname: Redmond
authors.forename: Janet M
authors.initials: JM
authors.lastname: Shaw
authors.forename: Ian J
authors.initials: IJ
authors.lastname: Hart
authors.forename: C Anthony
authors.initials: CA
authors.lastname: Hart
authors.forename: Rosalind L
authors.initials: RL
authors.lastname: Smyth
authors.forename: Malcolm G
authors.initials: MG
authors.lastname: Semple
mesh_terms: D002648:Child; D002675:Child, Preschool; D017931:DNA Primers / Q000235:genetics; D004739:England / Q000453:epidemiology; D006801:Humans; D015994:Incidence; D007223:Infant; D007231:Infant, Newborn; D029121:Metapneumovirus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D018184:Paramyxoviridae Infections / Q000175:diagnosis* / Q000453:epidemiology; D010802:Phylogeny; D012367:RNA, Viral / Q000235:genetics; D012141:Respiratory Tract Infections / Q000453:epidemiology / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D017385:Sequence Homology
chemical_list: D017931:DNA Primers; D015345:Oligonucleotide Probes; D012367:RNA, Viral