HPV in oral cavity: different methods for the detection of a large number of Alpha-PV and Beta-PV genera.

CHOUHY, DIEGO; PAOLINI, FRANCESCA; BOLATTI, ELISA; GIRI ADRIANA A; VENUTI, ALDO

Trieste

Encuentro; ICGEB DNA Tumour Virus Meeting; 2011

ICGEB

Introduction: Many reports indicate involvement of HPV, particularly the type 16, in a subset of head and neck squamous cell carcinoma (HNSCC). However, most of these studies were concentrated on the identification of only specific mucosal high-risk types of the genus alpha-papillomavirus. The aim of our study was to identify a broader spectrum of HPV types by different methods.    Materials and Methods: A total of 65 subjects were recruited and categorized in 3 groups:  pre-malignant lesions (group A), oral carcinomas (group B) and health controls (group C). All subjects were tested using 2 assays for mostly Alpha HPV detection. Both assays were based in the MY09/11 primer system but had different detection strategies: i) the L1HPVPCR assay that combines liquid hybridization with generic and specific probes and EIA for detection and typing and ii) HPV M based on the direct sequencing of the amplified products. In addition, all samples were further tested with two other assays for mostly beta HPV: HPV C, based on CP primer amplified products subjected to direct sequencing and the CUT primers system based on single tube “hanging droplet” nested PCR technique with original designed primer (Chouhy et al. Virology 2010) Results: Comparison between the assays for mucosal HPV detection had a general concordance of 65%. Only one patient was positive by the HPV M assay, corresponding to HPV-6. HPV infection was detected in 34% of patients by the L1HPVPCR assay, being the HPV-16 the most frequent type. On the other hand, HPV infection was detected in 57% of patients by the HPV C assay, being the HPV-20 the most frequent type. Data on the CUT assay will be disclosed. Conclusions: Despite the weak concordance among the utilized methods, mostly accounted for difference in the detection procedures (sequencing versus hybridization), nevertheless our data indicates the presence of beta and alpha HPV into the oral cavity. At the light of this observation, any further study on the HPV infection of oral cavity should be performed by methods able to detect large number of different mucosal and cutaneous HPV. Grants: MINCyT-MAE (IT/08/11)