title: Molecular characterisation of human hepatitis E virus from Italy: comparative analysis of five reverse transcription-PCR assays.
abstract: BACKGROUND
  Hepatitis E (HEV) is an important public-health concern as a major cause of enterically transmitted hepatitis worldwide. In industrialised countries it is considered rare, and largely confined to travellers returning from endemic areas. However, autochthonous (locally acquired) HEV infection is also emerging in these regions. The infection is caused by different genotypes, depending on whether it is travel-related or autochthonous. Conventional RT-PCR followed by sequencing of PCR products can identify HEV genotype and, depending on the region, the subtype, thus helping in defining the origin of infection and tracing the source of contamination.
  
  
  METHODS
  We re-analysed a collection of serum samples previously confirmed as hepatitis E positive by anti-HEV IgM and IgG assays as well as by Real-Time PCR, with the aim to compare the performances of five different broad range RT-PCR assays that could be provided for molecular characterisation of HEV. This approach is certainly valuable to investigate the molecular epidemiology of acute hepatitis E in countries where co-circulation of different genotypes occurs, like Italy.
  
  
  RESULTS
  Samples were analyzed by five assays targeting the ORF1, ORF2, and ORF2/3 regions. The sensitivity of these assays varied significantly, depending on the target region. Only 46% of samples tested positive by nested PCR; moreover, no single method was able to detect all positive samples. Most sequences originated from patients who had travelled to endemic areas (genotype 1), while the minority originated from Italian patients with no travel history (genotype 3).
  
  
  CONCLUSIONS
  Broad range methods for molecular characterization of HEV still need to be improved to detect all circulating strains.
authors.affiliation: Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy. giuseppina.larosa@iss.it.
authors.forename: Giuseppina
authors.initials: G
authors.lastname: La Rosa
authors.forename: Marta
authors.initials: M
authors.lastname: Fratini
authors.forename: Michele
authors.initials: M
authors.lastname: Muscillo
authors.forename: Marcello
authors.initials: M
authors.lastname: Iaconelli
authors.forename: Stefania
authors.initials: S
authors.lastname: Taffon
authors.forename: Michele
authors.initials: M
authors.lastname: Equestre
authors.forename: Paola
authors.initials: P
authors.lastname: Chionne
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Madonna
authors.forename: Giulio
authors.initials: G
authors.lastname: Pisani
authors.forename: Roberto
authors.initials: R
authors.lastname: Bruni
authors.forename: Anna Rita
authors.initials: AR
authors.lastname: Ciccaglione
mesh_terms: D016751:Hepatitis E / Q000453:epidemiology / Q000821:virology*; D016752:Hepatitis E virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007558:Italy / Q000453:epidemiology; D017720:Molecular Epidemiology / Q000379:methods; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
chemical_list: D012367:RNA, Viral
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