Cut primer system: an additional tool to study human papillomavirus infections in mucosal and cutaneous epithelia

CHOUHY, DIEGO; CERDA MB; MAMPRÍN D´ANDREA, R; GIRI ADRIANA A

Montreal

Conferencia; 26th International Papillomavirus Conference; 2010

International Papillomavirus Society

CUT primers are a new generic system targeting mucosal/cutaneous HPVs. In skin samples, CUT primers were able to identify viruses from the Alpha-PV (species 2, 3, 4, 8, 10), the Beta-PV (species 1, 2, 3), and the Gamma-PV (species 1, 2, 3) genera and lead to the characterization of HPV 115, a cutaneotropic Beta-PV species 3 virus. In order to investigate the ability of this primer system to detect mucosal HPVs, we analyzed 352 cervical swabs derived from women attending to Hospital-Escuela “Eva Perón”, Argentina. Pap smear screening scored 321 negative, 19 ASCUS, 10 LSIL and 2 HSIL. All samples were analyzed with CUT primers, a “hanging droplet PCR” amplification strategy and amplicon identification by direct sequencing. Results were compared with the L1HPVPCR assay that combines the widely used MY09/11 primers, liquid hybridization with generic and specific probes, and EIA for detection and typing of 16 HPVs (types 6, 11, 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 56, 58, 68, 73). Prevalences of HPV infection using CUT or MY primers were 37% and 32%, respectively, with a concordance of 75% (264/352) among both assays. The overall prevalence considering results from both systems was 47%. CUT primers identified 27 different viruses belonging to genus Alfa-PV from species 3, 5, 6, 7, 8, 9, 10, 13 and 15, all containing mucosal HPV. The most frequent types identified with CUT primers were types 66 (8%), 16 (6%), 52 (4%), cand89 (3%) while MY primers found types 16 (14%), 31 (6%), 56 (5%), 18 (3%) as the most prevalent ones. These data indicate that HPV epidemiology is complex and depends on the assays used for its characterization. In conclusion, CUT primer system provides an additional tool to study HPV infection in both mucosal and cutaneous samples. Funding. FIC-NIH(D43TW001037), ANPCyT (PICT 05-25464)