title: Molecular detection of hepatitis A virus in urban sewage in Rio de Janeiro, Brazil.
abstract: OBJECTIVE
  A one-year survey was conducted to examine hepatitis A virus (HAV) prevalence, distribution of genotypes and their relationship to bacterial indicators in raw and treated sewage samples.
  
  
  RESULTS
  Fifty sewage samples (raw = 25 and treated = 25) were collected twice monthly from one sewage treatment plant in Rio de Janeiro. Virus concentration was performed by adsorption to an electronegative membrane followed by ultrafiltration. Viral RNA was detected by nested reverse transcriptase-polymerase chain reaction (RT-PCR) and real-time PCR and positive products were directly sequenced. Total and faecal coliform concentrations were also determined. By nested RT-PCR, HAV RNA was detected in 16/50 (32%) and eight (16%) of them were found in treated sewage samples. By real-time PCR, HAV RNA was detected in 46/50 (92%) samples and 24 were from treated sewage. Phylogenetic analyses classified nine isolates (56%) as subgenotype IA and seven (44%) as IB.
  
  
  CONCLUSIONS
  Real-time PCR was more sensitive than nested RT-PCR; the presence of subgenotypes IA and IB was described and bacterial indicators cannot be used to predict HAV presence in sewage.
  
  
  CONCLUSIONS
  These results demonstrated that HAV still remains in the environment after sewage treatment and could play an important role in maintaining the endemicity of HAV infection.
authors.affiliation: Department of Virology, Oswaldo Cruz Institute, Rio de Janeiro, Brazil. lvillar@ioc.fiocruz.br
authors.forename: L M
authors.initials: LM
authors.lastname: Villar
authors.forename: V S
authors.initials: VS
authors.lastname: de Paula
authors.forename: L
authors.initials: L
authors.lastname: Diniz-Mendes
authors.forename: F R
authors.initials: FR
authors.lastname: Guimarães
authors.forename: F F M
authors.initials: FF
authors.lastname: Ferreira
authors.forename: T C
authors.initials: TC
authors.lastname: Shubo
authors.forename: M P
authors.initials: MP
authors.lastname: Miagostovich
authors.forename: E
authors.initials: E
authors.lastname: Lampe
authors.forename: A M C
authors.initials: AM
authors.lastname: Gaspar
mesh_terms: D001938:Brazil / Q000453:epidemiology; D004279:DNA, Viral / Q000032:analysis; D006506:Hepatitis A / Q000453:epidemiology / Q000635:transmission; D030041:Hepatitis A virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D012722:Sewage / Q000821:virology*; D014505:Urban Population
chemical_list: D004279:DNA, Viral; D012722:Sewage