title: Specific primer sets used to amplify by PCR the hepatitis B virus overlapping S/Pol region select different viral variants.
abstract: PCR detection of viral genomes has provided new insights into viral diagnosis. Nowadays, it is the most frequently used nucleic acid testing (qualitative and quantitative) technique. The aim of this study was to analyse the major circulating hepatitis B virus (HBV) variants PCR-amplified by three sets of primers in a patient infected with genotype E. The HBV S/Pol overlapping genomic region was amplified from the serum of an infected child using three primer sets previously described. Sequence analysis corresponding to the HBV S/Pol region revealed the presence of different viral populations depending on the set of primers used. D144A S-escape mutant was detected with two of the primer sets, while the rtL217R mutant within the Pol - conferring resistance to Adefovir - could be picked up with a different pair of primer sets. This study undoubtedly implies that the description of viral polymorphisms should be stated together with the sequence of the primers used for PCR amplification when studies of escape and/or antiviral-resistant HBV mutants are carried out.
authors.affiliation: Instituto de Microbiología y Parasitología Médica (IMPAM), Fac. de Medicina, Universidad de Buenos Aires (UBA),Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina.
authors.forename: M L
authors.initials: ML
authors.lastname: Cuestas
authors.forename: V L
authors.initials: VL
authors.lastname: Mathet
authors.forename: J R
authors.initials: JR
authors.lastname: Oubiña
mesh_terms: D017931:DNA Primers / Q000235:genetics*; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics*; D004259:DNA-Directed DNA Polymerase / Q000235:genetics; D005838:Genotype; D006509:Hepatitis B / Q000821:virology*; D006514:Hepatitis B Surface Antigens / Q000235:genetics; D006515:Hepatitis B virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D017422:Sequence Analysis, DNA
chemical_list: D017931:DNA Primers; D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens; D004259:DNA-Directed DNA Polymerase