title: Use of a universal virus detection assay to identify human metapneumovirus in a hematopoietic stem cell transplant recipient with pneumonia of unknown origin.
abstract: BACKGROUND
  Development of uncommon viral infections in immunocompromised transplant recipients can pose major diagnostic challenges. We present a case report of an immunocompromised patient suffering from pneumonia, for which the causative agent was not identified by routine methods.
  
  
  OBJECTIVE
  To identify the potential cause of the pneumonia using a degenerate oligonucleotide primer (DOP)-PCR assay that is designed to detect all viruses.
  
  
  METHODS
  DOP-PCR was applied to bronchoalveolar lavage fluid from this patient. Generic PCR products were cloned and sequenced.
  
  
  RESULTS
  The novel universal virus assay detected human metapneumovirus in the clinical sample. The finding was confirmed by two independent metapneumovirus specific PCRs targeting different regions of the viral genome.
  
  
  CONCLUSIONS
  The DOP-PCR was used to detect and identify the sequence of an unidentified virus. This study provides proof of concept for the use of clinically relevant specimens in this unbiased universal assay, which requires no previous viral sequence information.
authors.affiliation: Division of Viral Products, Center for Biologics, Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892-4555, USA.
authors.forename: Christine
authors.initials: C
authors.lastname: Uhlenhaut
authors.forename: Jeffrey I
authors.initials: JI
authors.lastname: Cohen
authors.forename: Daniel
authors.initials: D
authors.lastname: Fedorko
authors.forename: Santosh
authors.initials: S
authors.lastname: Nanda
authors.forename: Philip R
authors.initials: PR
authors.lastname: Krause
mesh_terms: D001483:Base Sequence; D001992:Bronchoalveolar Lavage Fluid / Q000821:virology; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects*; D006801:Humans; D016867:Immunocompromised Host; D029121:Metapneumovirus / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D018184:Paramyxoviridae Infections / Q000175:diagnosis*; D011014:Pneumonia / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA
chemical_list: D017931:DNA Primers