title: Genotyping of a nosocomial outbreak of pandemic influenza A/H1N1 2009.
abstract: BACKGROUND
  Epidemiological surveys have revealed outbreaks of pandemic influenza A (H1N1) 2009 in several different contexts. Molecular characterization of the influenza virus could help to provide a more accurate description of these outbreaks.
  
  
  OBJECTIVE
  To genotype pandemic influenza A (H1N1) 2009 isolates from an epidemiologically defined nosocomial outbreak.
  
  
  METHODS
  We sequenced the neuraminidase (NA) and hemagglutinin (HA) influenza A (H1N1) 2009 genes from ten HIV-positive patients involved in an epidemiologically defined outbreak in the Clinical Microbiology and Infectious Diseases (CMID) Department. Sequences were aligned to search for specific genetic features of the involved strain. We also analyzed 37 unrelated influenza A (H1N1) 2009 cases from other hospital departments. All the sequences were used to obtain phylogenetic trees.
  
  
  RESULTS
  Identical genotypic features were shared by nine of the 10 cases initially considered to be involved in the outbreak, but not by the remaining case. These features involved two silent mutations at N385 and V407 in the NA gene and three amino acid substitutions in the HA gene (D225E, A189T, and P300S). Searching for these substitutions in patients with influenza A (H1N1) 2009 hospitalized in other departments during the same period allowed us to identify an additional unsuspected immunocompetent case. The five outbreak-specific substitutions were absent in the remaining 36 unrelated controls. One of the substitutions (P300S) rendered detection of this variant by the CDC protocol inefficient. The other outbreak-specific substitutions (D225E and A189T) were identified at codons that have been analyzed in the context of virulence.
  
  
  CONCLUSIONS
  Genotyping is essential to ensure a more accurate description of pandemic influenza A (H1N1) 2009 outbreaks.
authors.affiliation: Servicio de Microbiología Clínica y Enfermedades Infecciosas, Hospital General Universitario Gregorio Marañón, Madrid, Spain.
authors.forename: B
authors.initials: B
authors.lastname: Rodríguez-Sánchez
authors.forename: M
authors.initials: M
authors.lastname: Alonso
authors.forename: P
authors.initials: P
authors.lastname: Catalán
authors.forename: M
authors.initials: M
authors.lastname: Sánchez Conde
authors.forename: F
authors.initials: F
authors.lastname: González-Candelas
authors.forename: M
authors.initials: M
authors.lastname: Giannella
authors.forename: E
authors.initials: E
authors.lastname: Bouza
authors.forename: D
authors.initials: D
authors.lastname: García de Viedma
mesh_terms: D019943:Amino Acid Substitution; D060085:Coinfection; D003428:Cross Infection / Q000150:complications / Q000453:epidemiology* / Q000821:virology; D004196:Disease Outbreaks; D005838:Genotype; D060005:Genotyping Techniques; D006679:HIV Seropositivity / Q000150:complications; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D007251:Influenza, Human / Q000150:complications / Q000453:epidemiology* / Q000821:virology; D008969:Molecular Sequence Data; D009154:Mutation; D009439:Neuraminidase / Q000235:genetics; D058873:Pandemics; D010802:Phylogeny; D014764:Viral Proteins / Q000235:genetics
chemical_list: D019267:Hemagglutinin Glycoproteins, Influenza Virus; D014764:Viral Proteins; C487630:NA protein, influenza A virus; D009439:Neuraminidase