Molecular characterisation and tissue tropism of a novel mupapillomavirus HPV204

BOSTJAN J. KOCJAN; LEA HOSNJAK; ANJA STERBENC; DIEGO CHOUHY; ELISA BOLATTI; ADRIANA A. GIRI; MARIO POLJAK

Lisboa

Conferencia; 30th International Papillomavirus Conference; 2015

International Papillomavirus Society

Human papillomaviruses (HPVs) represent a large family of DNA viruses,etiologically linked with various neoplasms of the skin and mucosa, taxonomically classified intogenera, species and types. Mupapillomavirus (Mu-PV) genus currently consists of only two HPVtypes: HPV1 and HPV63 identified in 1980 and 1993, respectively, both associated withsporadic cases of skin warts. In this study, a potentially novel Mu-PV type, designated asFJ947082, was fully cloned, sequenced and genetically characterized. The tissue predilection ofa novel type was assessed on a collection of skin, mucosal and mucocutaneous clinicalsamples. Methods: Partial FJ947082 L1-gene sequence, closely related to Mu-PVs, wasinitially obtained from a skin swab sample using CUT primers. Complete viral genome wasamplified from an anal swab sample of a 36-year old male patient with HPV53-positive analwarts using inverse long-range PCR, cloned into a plasmid vector, and sequenced by primerwalking. A quantitative FJ947082 type-specific real-time PCR was developed and used todetermine the prevalence of a novel HPV type in 100 tissue specimens of skin warts, 95 swabsof the anal canal (mainly from patients with anal warts), and 100 swabs of clinically normal oralmucosa. Phylogenetic evaluation, performed using a maximum-likelihood algorithm, was basedon the entire L1 gene sequence of FJ947082 and all currently officially recognized HPV types.Results: Novel Mu-PV genome comprises 7,227-bp (G+C content of 37.8%), and contains fiveearly (E1, E2, E4, E6 and E7) and two late genes (L1 and L2), and the non-coding regulatoryregion between L1 and E6 ORFs. Phylogenetic analysis confirmed that FJ947082 clusters tothe Mu-PV genus. With an L1 nucleotide sequence identity of 66.3% and 66.7% to HPV1 andHPV63, respectively, FJ947082 is classified as a new type in a new Mu-PV species Mu-3. Usingreal-time PCR, novel HPV was detected in 6/100 skin warts, 3/95 anal canal samples, and innone of the oral cavity samples. Conclusion: With full characterization of a third member of Mu-PV genus we expand the current knowledge of genetic diversity and tissue tropism of this PVgenus.