Identification and characterization of novel human papillomaviruses in oral rinse samples from oral cavity and oropharyngeal cancer patients.

OBJECTIVES: The objectives of this study were to: I) discover novel human papillomaviruses (HPVs) using next generation sequencing (NGS) technology in oral rinse samples collected from oral cavity cancer (OCC) and oropharyngeal cancer (OPC) patients; II) determine the prevalence of novel HPVs in archived OCC and OPC tissue samples; and III) examine the frequency of novel oncogenic HPVs in cancer and non-cancer oral rinse samples using real-time PCR. METHODS: Oral rinse samples were collected from 100 head and neck cancer patients, and 110 healthy individuals. NGS techniques were used to detect novel HPVs. RESULTS: Three potentially new types of HPV were discovered. Novel virus (NV) 14.4 was closely related to HPV76 with an 89% homology and is a member of the genus Beta-papillomavirus (ß-PV); NV69.1 was distantly related to the genus Alpha-papillomavirus (α-PV), and NV95 was closely related to HPV147 with a 65-77% homology and is part of the genus Gamma-papillomavirus (γ-PV). In archived oral tissue samples, NV14.4 was detected in a single patient with OCC. Of the oral rinse samples, NV69.1 was more prevalent than the other two NVs. CONCLUSIONS: Our results demonstrated that there are novel HPVs present in oral rinse samples that may be associated with OCC and OPC. These novel HPVs can be identified and characterized using NGS techniques.

HPV16 infection in oral cavity and oropharyngeal cancer patients.

Our objective was to compare archived tissue biopsy samples from the oral cavity and oropharynx, in terms of human papillomavirus (HPV) 16 infection. We used Taqman real-time PCR assay to detect HPV16 in 121 archived biopsy samples from the oral cavity and 100 samples from the oropharynx. Among patients with oral cavity cancer (OCC), 9% (6/65) had HPV16 infection which was significantly less than those with oropharyngeal cancer (OPC) where 79% (39/50) were HPV16 positive (P < 0.001). Our results demonstrated a significant difference between genders where males had a seven times higher chance of having HPV16 infection (P < 0.001). Viral load variation between each group was demonstrated. The median viral load in OPC was similar in OCC cases, but cancer samples were significantly higher than in non-cancer cases (oral cavity samples P = 0.015; oropharynx samples P = 0.09). From our results, we conclude that there is a significant difference in HPV16 detection between OCC and OPC, and HPV16 differs greatly between male and female cancer patients. (J Oral Sci 58, 265-269, 2016).

Detection of HPV in oral rinse samples from OPSCC and non-OPSCC patients.

BACKGROUND: Due to the increasing rates of oropharyngeal cancer, oral HPV infection is a significant concern. Methods for detecting oral HPVs is not standardized as there are different techniques available. We propose that use of oral rinse samples to detect for HPVs is a suitable technique within a clinic setting. Thus, our main objective is to study HPV detection in oral rinse samples. METHODS: In our study, we used oral rinse sample collection coupled with real-time PCR to detect for HPVs types 16 and 18, and preferentially amplified FAP PCR samples to detect for a broad range of HPVs, in oropharyngeal squamous cell carcinoma (OPSCC), non-OPSCC, and healthy patients. RESULTS: Thirty three percent of 100 cancer patients were positive for any type of HPV; of those 23 were positive for HPV16. Only 1 of 110 healthy controls was positive (this subject was positive for HPV18). CONCLUSION: Our results indicate that HPV detection in oral rinse samples may be useful as a screening tool to detect HPV-associated oral cancers.

Characterization of three novel human papillomavirus types isolated from oral rinse samples of healthy individuals.

BACKGROUND: Despite the strong evidence of HPV infection as the etiological agent in a subset of oral cancer, oral α-HPV detection is rare in healthy individuals, and little is known of the existing of novel HPV types in oral cavity. OBJECTIVE: We determined whether novel HPV types can be isolated from oral rinse samples collected from healthy individuals. STUDY DESIGN: We performed rolling circle amplification (RCA) coupled with degenerated PCR assay on 48 oral rinse samples to amplify novel HPV types. Full length HPV DNA was cloned using long range PCR. Quantitative type specific Taqman assays were used to determine the prevalence of novel HPV types in 158 archived oral tissue samples. RESULTS: We were able to isolate four novel human papillomavirus types. Full length HPV DNA was cloned for three of the four novel HPV types. All four HPV types belong to the genus Gammapapillomavirus (γ-PV), where HPV 171 is most closely related to HPV 169, showing 88% similarity; HPV 172 is most closely related to HPV 156, showing 70% similarity; HPV 173 is most closely related to HPV 4, showing 73% similarity; oral sample lavage (OSL) 37 is most closely related to HPV 144, showing 69% similarity. Finally, we showed that HPV 173 was rarely present in oral tissues (2/158), HPV 172 was only detected in normal oral tissues (25/76), and HPV 171 was more prevalent in malignant oral tissues (17/82 vs. 10/76, p=0.21). CONCLUSIONS: Novel γ-HPV types are present in oral cavity of healthy individuals.