RESUMO
A modified pan-PV consensus-degenerate hybrid oligonucleotide primer (CODEHOP) PCR was developed for generic and sensitive detection of a broad-spectrum of human papillomaviruses (HPVs) infecting the cutaneous epithelium. To test the analytical sensitivity of the assay we examined 149 eyebrow hair follicle specimens from immunocompetent male patients. HPV DNA was detected in 60â% (89/149) of analysed eyebrow samples with a total of 48 different HPV sequences, representing 21 previously described HPVs and 27 putative novel HPV types. Evidence for ten novel HPV subtypes and seven viral variants, clustering to three out of five genera containing cutaneous HPVs, was also obtained. Thus, we have shown that the modified pan-PV CODEHOP PCR assay is able to identify multiple HPV types, even from different genera, in the same clinical sample. Overall, these results demonstrate that the pan-PV CODEHOP PCR is an excellent tool for screening and identification of novel cutaneous HPVs, even in samples with low viral loads.
Assuntos
Betapapillomavirus/isolamento & purificação , Primers do DNA/química , DNA Viral/genética , Gammapapillomavirus/isolamento & purificação , Genótipo , Infecções por Papillomavirus/diagnóstico , Reação em Cadeia da Polimerase/métodos , Adulto , Sequência de Bases , Betapapillomavirus/classificação , Betapapillomavirus/genética , Primers do DNA/metabolismo , Sobrancelhas/virologia , Gammapapillomavirus/classificação , Gammapapillomavirus/genética , Folículo Piloso/virologia , Humanos , Masculino , Tipagem Molecular/métodos , Infecções por Papillomavirus/epidemiologia , Infecções por Papillomavirus/virologia , Filogenia , Prevalência , Sensibilidade e Especificidade , Eslovênia/epidemiologiaRESUMO
We evaluated the concordance between ß-HPVs detected in external genital skin, anal canal, and oral cavity specimens collected simultaneously from 717 men that were participating in the multinational HIM Study. Viral genotyping was performed using the Luminex technology. Species- and type-specific concordance was measured using kappa statistics for agreement. Overall, concordance of ß-HPVs across sites was low and mainly observed among paired genital/anal canal samples. When grouped by species, solely ß-4 HPVs showed moderate concordance in genital/anal pairs (κ = 0.457), which could be attributed to the substantial concordance of HPV-92 in men from Brazil and Mexico (κ > 0.610). ß-HPV type concordance was higher in Mexico, where HPV-19 was consistently concordant in all anatomic site combinations. Our analysis indicates that type-specific concordance across sites is limited to few viral types; however, these infections seem to occur more often than would be expected by chance, suggesting that although rare, there is agreement among sites.
Assuntos
Canal Anal/virologia , Betapapillomavirus/classificação , Betapapillomavirus/isolamento & purificação , Genitália Masculina/virologia , Genótipo , Mucosa Bucal/virologia , Infecções por Papillomavirus/virologia , Betapapillomavirus/genética , Brasil , Florida , Técnicas de Genotipagem , Homossexualidade Masculina , Humanos , Masculino , MéxicoRESUMO
We present the first longitudinal study reporting the natural history of human papillomavirus (HPV) infection in sun-exposed skin of healthy individuals living in a geographical area in which solar UV radiation is influenced by the ozone content of the atmosphere. During three climatic seasons, skin swab samples were obtained from 78 healthy individuals and the prevalence of cutaneous HPVs was assessed with broad-spectrum FAP and CUT primers and determined at 54, 45 and 47â% in spring, summer and winter, respectively. Frequencies of mixed HPV infections were significantly higher in spring with respect to summer and winter (P=0.02). Seventy-one different HPV types/putative types were identified. While 62 volunteers were HPV-infected in at least one season, 23 had persistent infections. ß-PVs (ß-1) were the most prevalent and persistent. Age was associated with both the infection status (P=0.01) and the type of HPV infection (no infection, indeterminate/transient, persistent P=0.02). The molecular/phylogenetic analysis of the newly identified ß-PV, officially designated as HPV209, showed that the virus has a typical genomic organization of cutaneous HPVs with five early (E6, E7, E1, E2 and E4) and two late genes (L2 and L1), which clusters to the species ß-2. This provides useful data on cutaneous HPV infections in high UV-exposed regions.