Development and Validation of a Novel and Rapid Molecular Detection Method for High-Risk Human Papillomavirus in Formalin-Fixed, Paraffin-Embedded Tumor Tissue.

The most widely applied algorithm for human papillomavirus (HPV) detection in formalin-fixed, paraffin-embedded (FFPE) specimens of oropharyngeal head and neck squamous cell carcinoma (HNSCC) consists of p16INK4A immunostaining followed by PCR-based detection of high-risk HPV DNA on p16INK4A-immunopositive samples. However, in nonoropharyngeal HNSCC this algorithm fails, hampering correct interpretation of the prevalence and prognosis of HPV in these cases. In this study, we developed and validated a molecular HPV detection method for FFPE specimens of oropharyngeal and nonoropharyngeal HNSCC. Sectioning of FFPE blocks was circumvented by using punch biopsies from tumor-enriched regions of FFPE tissue blocks, and combined extraction was applied to obtain high-quality DNA and RNA from the punch biopsy. Next, PCR-based detection of HPV DNA was performed for 15 high-risk HPV types with subsequent detection of E6 mRNA for validation. The combined DNA/RNA FFPE test of tissue cores was assessed in well-characterized cohorts with known HPV status based on earlier work, that is, a cohort of oropharyngeal HNSCC (n = 80) and oral cavity HNSCC (n = 25), and reached an accuracy of 97% and 100%, respectively. In conclusion, our method is rapid, simple, and shows an excellent diagnostic performance for detection of HPV type 16. Ultimately, it can be applied for large cohort studies to determine the etiologic fraction and prognostic implication of HPV in nonoropharyngeal HNSCC.

Prevalence of Neovaginal High-Risk Human Papillomavirus Among Transgender Women in The Netherlands.

BACKGROUND: Worldwide, transgender women are a high burden population for sexually transmitted diseases. Neovaginal high-risk human papillomavirus (hrHPV) infection has been documented among transgender women, but its prevalence remains unclear. The objective of this study was to determine the prevalence of neovaginal hrHPV in a sample of transgender women in The Netherlands. METHODS: Between June 2015 and December 2015, neovaginal samples were obtained from all transgender women who underwent vaginoplasty and attended our outpatient clinic for postoperative follow-up at least 1 year after surgery. High-risk HPV DNA detection and partial genotyping was performed by the HPV-risk assay. Genotyping of non-16/18-hrHPV-positive samples was subsequently performed by GP5+/6+-PCR followed by Luminex suspension array technology. Physical examination and standardized (sexual) history taking was conducted. RESULTS: Valid neovaginal swabs were obtained from 54 transgender women (median age, 40.7 years [range, 19.2-60.3]; median postoperative time, 2.4 years [range, 1.0-34.2]). No transgender women were employed in the commercial sex industry. Of 28 sexually active transgender women, 6 (20%) tested positive for neovaginal hrHPV including types 16, 45, 51, 59, 66, and X. There were no concomitant neovaginal lesions nor neovaginal symptoms. All sexually inactive transgender women tested negative for neovaginal hrHPV. CONCLUSIONS: A prevalence of neovaginal hrHPV infection of 20% is observed in Dutch transgender women, who self-reported to be sexually active. The clinical consequences neovaginal hrHPV infection in transgender women require further attention.

Sexual behaviour and less frequent bathing are associated with higher human papillomavirus incidence in a cohort study of uncircumcised Kenyan men.

OBJECTIVES: Data on the acquisition of human papillomavirus (HPV) infection in men are limited, especially from developing regions including Africa. The objective of this study was to characterise and determine the risk factors of HPV acquisition among a cohort of uncircumcised men participating in a randomised controlled trial (RCT) of male circumcision in Kisumu, Kenya. METHODS: Penile exfoliated cell specimens were collected at baseline, 6- and 12-month follow-up visits from the glans/coronal sulcus and shaft of men enrolled in the control arm of the RCT between 2002 and 2005. All participants were HIV seronegative, aged 17-24 years at baseline and remained uncircumcised over follow-up. Specimens were tested with GP5+/6+ PCR to detect 44 HPV types. Parametric frailty models were used to assess risk factors of HPV incidence. RESULTS: The median age of 966 participants was 20 years. The median follow-up time was 12.1 months. The incidence rate (IR) of any HPV infection was 49.3/1000 person-months with HPV16 having the highest IR (10.9/1000 person-months). The strongest risk factors for overall HPV incidence were bathing less frequently than daily (adjusted HR=2.6; 95% CI 1.0 to 6.5) and having ≥ 2 female sexual partners in the past year (adjusted HR=1.6; 95% CI 1.2 to 2.1). CONCLUSIONS: HPV IRs were notably high in this cohort of high-risk, uncircumcised men from Kisumu, Kenya, with the number of sexual partners and bathing frequency being the strongest risk factors.

Prevalence and risk factors of human papillomavirus infection by penile site in uncircumcised Kenyan men.

Human papillomavirus (HPV) prevalence was estimated from 2,705 sexually active, uncircumcised, human immunodeficiency virus seronegative men aged 17-28 years in Kisumu, Kenya. HPV prevalence was 51.1% (95% confidence interval: 49.2-53.0%) in penile cells from the glans/coronal sulcus and/or shaft. HPV prevalence varied by anatomical site, with 46.5% positivity in the glans/coronal sulcus compared with 19.1% in the shaft (p < 0.0001). High-risk HPV was detected in 31.2% of glans and 12.3% of shaft samples (p < 0.0001). HPV16 was the most common type and 29.2% of men were infected with more than one HPV type. Risk factors for HPV infection included presence of C. trachomatis, N. gonorrhea, self-reported sexually transmitted infections, and less frequent bathing. Lifetime number of sexual partners and herpes simplex virus type-2 seropositivity were also marginally associated with HPV infection.