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Identification of human papillomavirus type 156, the prototype of a new human gammapapillomavirus species, by a generic and highly sensitive PCR strategy for long DNA fragments.
Chouhy, Diego; Bolatti, Elisa M; Piccirilli, Gustavo; Sánchez, Adriana; Fernandez Bussy, Ramón; Giri, Adriana A.
Afiliação
  • Chouhy D; Instituto de Biología Molecular y Celular de Rosario (CONICET), Rosario, Argentina.
  • Bolatti EM; Area Virología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, Argentina.
  • Piccirilli G; Instituto de Biología Molecular y Celular de Rosario (CONICET), Rosario, Argentina.
  • Sánchez A; División de Dermatología, Hospital Provincial del Centenario, Rosario, Argentina.
  • Fernandez Bussy R; División de Dermatología, Hospital Provincial del Centenario, Rosario, Argentina.
  • Giri AA; División de Dermatología, Hospital Provincial del Centenario, Rosario, Argentina.
J Gen Virol ; 94(Pt 3): 524-533, 2013 Mar.
Article em En | MEDLINE | ID: mdl-23136368
This study developed a hanging-droplet long PCR, a generic and highly sensitive strategy to facilitate the identification of new human papillomavirus (HPV) genomes. This novel procedure used for the first time the hanging-droplet PCR technique for the amplification of long DNA fragments with generic primers targeting the L1 and E1 regions. It was first applied to the amplification of types belonging to the highly divergent genus Gammapapillovirus (γ-PV). The hanging-droplet long PCR was 100-fold more sensitive than a simple long PCR procedure, detecting as few as ten copies of HPV-4. Nineteen skin samples, potentially containing putative HPV types from the γ-PV genus, were also screened. The method identified four γ-PV genomic halves from new and previously described putative types, and made the full characterization of HPV-156 possible. This novel virus meets the criteria for a new species within the γ-PV genus, with nucleotide identities in the L1 ORF ranging from 58.3 to 67.3 % compared with representative types of the current γ-PV species. HPV-156 showed the highest identity to HPV-60 (67.3 %) from species γ-4, and was consistently closely related to it in both late- and early-gene-derived phylogenies. In conclusion, this report provides a versatile and highly sensitive approach that allowed identification of the prototype of a new species within the γ-PV genus. Its application with primers targeting the different genera in which both human and non-human PVs are distributed may facilitate characterization of the missing members of the family Papillomaviridae.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: DNA Viral / Reação em Cadeia da Polimerase / Alphapapillomavirus / Gammapapillomavirus Tipo de estudo: Diagnostic_studies / Prognostic_studies Limite: Humans Idioma: En Revista: J Gen Virol Ano de publicação: 2013 Tipo de documento: Article País de afiliação: Argentina País de publicação: Reino Unido
Texto completo
Adicionar na Minha BVS
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XML
PubMed Links
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: DNA Viral / Reação em Cadeia da Polimerase / Alphapapillomavirus / Gammapapillomavirus Tipo de estudo: Diagnostic_studies / Prognostic_studies Limite: Humans Idioma: En Revista: J Gen Virol Ano de publicação: 2013 Tipo de documento: Article País de afiliação: Argentina País de publicação: Reino Unido