Abstract
Oral infection by Human Papillomavirus (HPV) has recently gained great attention because of its involvement in the development of a subset of head and neck squamous cell carcinoma. The role of specific Alpha-HPVs in this regard has been well established, whereas the contribution of other genera is under investigation. Despite their traditional classification as “cutaneous” types, Beta and Gamma HPVs are frequently detected in oral samples. Due to the lack of a standardized protocol, a large variety of methodologies have been used for oral sample collection, DNA extraction, HPV detection and genotyping. Laboratory procedures influence the evaluation of oral HPV prevalence, which largely varies also according to the population characteristics, e.g., age, gender, sexual behavior, Human Immunodeficiency Virus (HIV) status. Nevertheless, oral infection by Beta and Gamma HPVs seems to be even more common than Alpha-HPVs. The latter is 5–7% in the general population, and increases up to 30% approximately in HIV-infected men who have sex with men. Despite major advances in the evaluation of oral HPV prevalence, its natural history is still little understood, especially for Beta and Gamma HPVs. The latest technologies, such as Next Generation Sequencing (NGS), can be exploited to gain new insights into oral HPV, and to improve the identification of novel HPV types.
Highlights
Human Papillomaviruses (HPVs) cause the most widespread sexually transmitted infection [1]
Laboratory procedures influence the evaluation of oral HPV prevalence, which largely varies according to the population characteristics, e.g., age, gender, sexual behavior, Human Immunodeficiency Virus (HIV) status
Based on variations in the sequence of the L1 open reading frame, which encodes for the major capsid protein, HPVs are classified into five genera (Alphapapillomavirus, Alpha-HPV; Betapapillomavirus, Beta-HPV; Gammapapillomavirus, Gamma-HPV; Mupapillomavirus, mu-HPV; Nupapillomavirus, nu-HPV) [2] but the majority of them belong to Alpha-HPV, Beta-HPV and Gamma-HPV genera
Summary
Human Papillomaviruses (HPVs) cause the most widespread sexually transmitted infection [1]. This includes the following steps: (1) oral sample collection (followed by a pre-analytical phase); (2) nucleic acid extraction and purification; (3) HPV-DNA amplification, detection and genotyping. Oral HPV Infection by Alpha, Beta and Gamma HPVs: the Methodological Aspects The laboratory workflow for the assessment of oral HPV infection is depicted in Fig-4 of 21 ure 1 This primer set targets a 65 base pair fragment of L1 region and is commercially available in two different HPV tests: (i) the original SPF10-DNA enzyme immunoassay (DEIA)-LIPA25 system, where a DEIA performed in a 96-well format is followed by a reverse hybridization for 25 HPVs [43]; (ii) a reverse blotting hybridization system, which is based on hybridization of the biotinylated amplicons with probes immobilized on nitrocellulose strips and detects up to 32 HPV genotypes, depending on the kit version (INNO-LiPA HPV Genotyping, Innogenetics/Fujirebio, Göteborg, Sweden)
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