Human Papillomavirus DNA Polymerase Chain Reaction Research Articles

Advances in testing for human papillomavirus -mediated head and neck cancer.

New evidence has recently emerged regarding the utility and benefits of dual p16 INKa (p16) and Human papillomavirus (HPV) status testing when determining the diagnosis and prognosis of patients with oropharyngeal cancer. HPV RNA polymerase chain reaction (PCR) is the most accurate diagnostic test. The other assays (HPV DNA PCR, HPV DNA/RNA in-situ hybridization (ISH) and p16) applied to formalin fixed tumour tissue have varying but high sensitivities and specificities. Dual p16 and HPV testing identifies discordant (p16+/HPV- or p16-/HPV+) results in 9.2% of cases, who have significantly poorer prognoses than p16+/HPV+, particularly in smokers. The proportion of discordant cases varies by region, and appears to be highest in regions with lowest attributable (p16+/HPV+) fractions. Dual testing improves prognostication for oropharyngeal cancer cases by identifying discordant cases and improving the prognostic power of the Tumour Node Metastasis (TNM) classification, especially in regions with high discordant rates. Dual testing is essential when considering patients for clinical trials of treatment de-escalation, and may be important when counselling patients on prognosis, especially in regions with high discordant rates and in smokers.

Radiomic Analysis for Human Papillomavirus Assessment in Oropharyngeal Carcinoma: Lessons and Pitfalls for the Next Future

Background: Oropharyngeal Squamous Cell Carcinoma (OPSCC) is rapidly increasing due to the spread of Human Papillomavirus (HPV) infection. HPV-positive disease has unique characteristics, with better response to treatment and consequent better prognosis. HPV status is routinely assessed via p16 immunohistochemistry or HPV DNA Polymerase Chain Reaction. Radiomics is a quantitative approach to medical imaging which can overcome limitations due to its subjective interpretation and correlation with clinical data. The aim of this narrative review is to evaluate the impact of radiomic features on assessing HPV status in OPSCC patients. Methods: A narrative review was performed by synthesizing literature results from PUBMED. In the search strategy, Medical Subject Headings (MeSH) terms were used. Retrospective mono- or multicentric works assessing the correlation between radiomic features and HPV status prediction in OPSCC were included. Selected papers were in English and included studies on humans. The range of publication date was July 2015–April 2023. Results: Our research returned 23 published papers; the accuracy of radiomic models was evaluated by ROC curves and AUC values. MRI- and CT-based radiomic models proved of comparable efficacy. Also, metabolic imaging showed crucial importance in the determination of HPV status, albeit with lower AUC values. Conclusions: Radiomic features from conventional imaging can play a complementary role in the assessment of HPV status in OPSCC. Both primary tumor- and nodal-related features and multisequencing-based models demonstrated higher accuracy.

Oropharyngeal squamous cell carcinoma: p16/p53 immunohistochemistry as a strong predictor of HPV tumour status.

Oropharyngeal squamous cell carcinomas (OPSCC) related to human papillomavirus (HPV) infection have a better prognosis than those without HPV infection. Although p16INK4a overexpression is used as a surrogate marker for HPV infection, 5-20% of p16-positive OPSCC are described as being unrelated to HPV infection, with worse overall survival compared to OPSCC-related HPV. There is therefore a risk of undertreating a proportion of OPSCC patients falsely considered to be HPV-driven because of p16 positivity. TP53 mutations are highly prevalent in OPSCC driven by mutagens in tobacco and alcohol. We describe herein a combined p16/p53 algorithm to predict HPV tumour status in OPSCC. A total of 110 OPSCC were identified in the database of the pathology department and were studied using p16 and p53 immunohistochemistry. For p16-positive or p16-negative/wild-type patterns-p53 (WT-p53) cases (n=63), DNA in-situ hybridisation for high-risk HPV was performed, and if negative the HPV status was controlled by HPV DNA polymerase chain reaction (PCR) (n=19). A significant association between TP53 mutation and pattern of p53 expression was found (WT-p53, seven of 16, P<0.001). The p16-positive/WT-p53 was significantly associated with HPV+ tumour status (p16-positive/WT-p53, 50 of 110, P<0.001). Interestingly, a subset of p16-positive OPSCC was unrelated to HPV (13.5%, eight of 59), and showed mutant-type staining of p53 expression. The p16 protein immunopositivity in conjunction with the mutant-type pattern of p53 staining helped to reclassify a subset of p16-positive OPSCC as OPSCC-unrelated HPV. This approach could be routinely applied by pathologists involved in the management of OPSCC, because of their potential therapeutic implications.

Correlation of p16 immunostaining in cell-blocks with corresponding tissue specimens for squamous cell carcinomas of the oropharynx.

The goal of this study was to evaluate the performance of p16 staining in cell-blocks vs tissue specimens as a surrogate marker for human papillomavirus (HPV) status in oropharyngeal squamous cell carcinomas. Head and neck squamous cell carcinoma cases presenting as a neck mass with a p16 result on cytology and corresponding tissue specimens (1 January 2014 to 30 June 1920) were included in the study. The following were assessed from cell-block material: number of tumour clusters, percentage of tumour cells with p16 staining, and presence of staining in clusters vs single cells. Results were compared to tissue p16 status. Results of any other ancillary HPV testing were also noted. Forty-two head and neck squamous cell carcinoma neck metastases (35 oropharyngeal, five non-oropharyngeal, and 2 unknown primaries) were identified. The p16 staining pattern in cell-blocks was seen in single cells (27.6%), clusters (44.8%), or both (27.6%). The percentage of tumour cells staining for p16 in cell-blocks was much lower than in corresponding tissue specimens. There were four false negatives and one false positive (concurrent HPV DNA polymerase chain reaction testing was positive in cytology and surgical material). Compared to tissue, the cut-off for p16 interpretation in cell-blocks is substantially lower and staining may be present in single cells or clusters. In 96.9% of cases, any p16 staining in cell-blocks correlated with positive p16 staining in surgical specimens. However, a negative or discrepant p16 result on cell-block should prompt confirmatory HPV studies, as false negative p16 staining in cell-blocks is high.

Expanding the Morphologic, Immunohistochemical, and HPV Genotypic Features of High-grade Squamous Intraepithelial Lesions of the Vulva With Morphology Mimicking Differentiated Vulvar Intraepithelial Neoplasia and/or Lichen Sclerosus.

Squamous cell carcinoma of the vulva can arise through 2 pathways: human papillomavirus (HPV)-dependent high-grade squamous intraepithelial lesions (previously termed usual vulvar intraepithelial neoplasia) or HPV-independent (differentiated vulvar intraepithelial neoplasia, dVIN). Distinguishing between the 2 types can be clinically and histologically difficult. A subset of high-grade squamous intraepithelial lesions with superimposed chronic inflammation mimicking dVIN has recently been reported; p53 shows characteristic mid-epithelial staining (with basal sparing) in such cases. The pathology databases of 2 academic institutions were searched for vulva specimens with corresponding p53 and p16 immunohistochemical stains, yielding 38 specimens (from 27 patients). In situ hybridization and multiplex polymerase chain reaction-MassArray for high-risk HPV were performed on at least 1 block from each patient. All cases resembled dVIN or lichen sclerosus morphologically, but with a higher degree of atypia. All but 1 case demonstrated mid-epithelial p53 staining with basal sparing by immunohistochemistry. All cases showed block positivity for p16 and at least patchy positivity by HPV in situ hybridization. Of the 23 cases with valid HPV DNA polymerase chain reaction results, 15 were positive and 8 were negative. Of the positive cases, HPV16 was identified in 10 cases, with other high-risk types in the remaining 5. To our knowledge, this is the largest cohort of high-grade squamous intraepithelial lesions mimicking dVIN reported to date. Prior studies reported positivity for HPV16 in all cases tested, however, we found HPV16 in only 67% of HPV positive cases. This case series highlights the importance of immunohistochemistry, and occasionally HPV in situ hybridization, for accurate diagnosis, and expands the spectrum of associated HPV types.

Long-term outcomes in patients with recurrent human papillomavirus-positive oropharyngeal cancer after upfront transoral robotic surgery.

Clinical course following failure of human papillomavirus (HPV)-positive oropharyngeal cancers (HPV + OPC) is poorly understood. This study aims to characterize disease course following failure after transoral robotic surgery (TORS). We identified patients with HPV + OPC-treated upfront with TORS at our institution from 2007 to 2017. HPV status was confirmed with immunohistochemistry or HPV DNA polymerase chain reaction. Patient characteristics, treatment modalities, and post-recurrence outcomes were analyzed for the recurrent cohort. Of the 317 HPV + OPC patients, 28 (8.8%) experienced recurrence, all of HPV 16/18 subtypes. Median post-recurrence survival was 19.8 months (range 2.3-195.8 months) in the 12 locoregional and 16 months (range 2.4-79.5 months) in the 14 distant failures. Sixteen are alive with a median of 39.8 months (range 5.5-209.4 months) after retreatment. This is one of the largest series evaluating survival following TORS failure in HPV + OPC. Despite failure, long-term survival and durable remission are possible with single-modal or multiple-modal salvage treatment.

In situ hybridization for high-risk HPV E6/E7 mRNA is a superior method for detecting transcriptionally active HPV in oropharyngeal cancer

Current human papillomavirus (HPV) detection methods in oropharyngeal squamous cell carcinoma (OPSCC) have varying sensitivity and specificity. We aimed to compare different HPV-detection methods against the test used in clinical practice, ie, p16 immunohistochemistry (IHC) and to evaluate whether another HPV-detection test additional to p16 IHC would be worthwhile in OPSCC specimens. The study cohort comprised 357 consecutive OPSCC patients during two time periods: 2000-2009 and 2012-2016. From tumor tissue slides, HPV mRNA via in situ hybridization (ISH), HPV DNA via ISH and HPV DNA via polymerase chain reaction (PCR) were detected. The results of these methods were compared with p16 IHC results. Additionally, clinicopathological factors were compared with the methods studied. The sensitivity of HPV mRNA ISH, HPV DNA ISH and HPV DNA PCR were 93.4%, 86.3%, and 83.5%, respectively. The corresponding specificity was 92.4%, 95.3%, and 89.1%, respectively. The negative predictive value for p16 IHC was highest (89.0%) when using mRNA ISH, and followed by DNA ISH (83.5%). ISH for high-risk HPV E6/E7 mRNA was found to be a highly specific and sensitive method for detecting HPV in OPSCC. As p16 protein may be overexpressed due to HPV-independent mechanisms, all p16 IHC-positive OPSCCs should be considered for retesting using mRNA ISH in order to verify transcriptionally active HPV. This is especially critical when considering de-escalated treatment approaches for patients with HPV-positive tumors and still maintaining favorable outcomes for this subgroup of patients.

Role of the human papillomavirus in malignant transformation of oral leukoplakia distinct from oropharyngeal squamous cell carcinoma: A study of 76 patients with internal-control specimens

This study sought to investigate the role of human papillomavirus (HPV) in the carcinogenesis of oral leukoplakia (OLK), with the oral cavity as the site of interest. A total of 76 patients (152 specimens) were enrolled in the study. The patients were divided into 2 groups: the malignant transformation of OLK (OLK-MT) group and the non-malignant transformation of OLK (OLK-non-MT) group. HPV reverse dot blot analysis, HPV DNA polymerase chain reaction (PCR), and p16INK4A immunohistochemistry (IHC) were used to determine HPV infection status. Transformation of OLK commonly occurred in the lateral/ventral tongue, buccal mucosa, and gingiva. On the basis of the initial analysis of specimens, only 5.3% (4 of 76) of patients were found to be HPV-16 positive, and these patients' final specimens yielded negative results. Overexpression of p16INK4A in the dysplastic stage was associated with the transformation of OLK (P = .013; odds ratio = 3.544). Transformation of OLK was common in patients who are elderly, in females, and in nonsmokers/nondrinkers; lesions were located in the lateral/ventral tongue, with dysplasia and overexpressed p16INK4A seen during the initial stage. HPV may be an opportunistic infection in the oral cavity and may not be a cause of malignant transformation of OLK. p16INK4A expression, which initially increases and then diminishes or disappears, may be an early predictor of malignant transformation.

Role of Conventional Pap’s Smear in Diagnosis of Atypical Squamous Cell-Indeterminate Significance (ASC-US) and Low-Grade Squamous Intraepithelial Lesion (LSIL) in Comparison with Human Papillomavirus PCR Test

Human papillomavirus (HPV) infection is most common etiological agent of carcinoma cervix. Most prevalent carcinogenic HPV types are 16 and 18. Cervical lesion can be detected by conventional Pap’s smears and polymerase chain reaction (PCR). Molecular techniques are costly and not available everywhere. The aim of study was to evaluate efficacy of conventional Pap’s smear in comparison with HPV PCR for diagnosis of ASC-US (atypical squamous cell-indeterminate significance) and LSIL (low-grade squamous intraepithelial lesion). We prospectively collected data of conventional Pap’s smears from May 2017 to September 2017. Pap’s smears showing epithelial cell abnormality were sent for HPV DNA PCR as per Bethesda guidelines 2014. Total 252 Pap’s smears were collected. Age range of affected women was 21–45 years. Chief complaints of patients were discharges, pain, and bleeding per vagina. Out of 252 Pap’s smears, 47 Pap’s smears showed epithelial cell abnormalities. The abnormalities included ASC-US in 59.5%, LSIL in 17.0%, ASC-H (cannot exclude HSIL) in 10.6%, HSIL (high-grade squamous intraepithelial lesion) and AGC in 4.2% each, adenocarcinoma in situ and squamous cell carcinoma in 2.1% each. Out of 36 ASC-US and LSIL abnormal Pap’s smears, six patient’s (five ASC-US and one LSIL) samples were sent for HPV DNA PCR. All six patients were negative for HPV DNA on PCR. Early-stage lesions of cervix may be detected by Pap’s smear even in PCR-negative cases. Conventional Pap’s smear is cost-effective method for detection of cervical lesions like ASC-US and LSIL.

Refining prognostic stratification of human papillomavirus-related oropharyngeal squamous cell carcinoma: different prognosis between T1 and T2.

PurposeTo validate the 8th edition of the American Joint Committee on Cancer/Union for International Cancer Control (AJCC/UICC) TNM staging system for human papillomavirus (HPV)-related oropharyngeal squamous cell carcinoma (OPSCC) and investigate whether a modified classification better reflects the prognosis.Materials and MethodsMedical records of patients diagnosed with non-metastatic HPV-related OPSCC between 2010 and 2016 at a single institution were retrospectively reviewed. HPV status was determined by immunohistochemical analysis of p16 and/or HPV DNA polymerase chain reaction (PCR). We reclassified TNM stage T0-1 and N0-1 as group A, T2-3 or N2 as B, and T4 or N3 as C. Survival analysis according to 8th AJCC/UICC TNM staging and the modified classification was performed.ResultsOf 383 OPSCC patients, 211 were positive for HPV DNA PCR or p16. After exclusion, 184 patients were included in this analysis. Median age was 56 years (range, 31 to 81 years). Most primary tumors were in the palatine tonsil (148 tumors, 80%). The eighth AJCC/UICC TNM classification could not differentiate between stage I and II (p = 0.470) or II and III (p = 0.209). Applying modified grouping, the 3-year overall survival rate of group A was significantly higher than that of group B and C (98% vs. 91%, p = 0.039 and 98% vs. 78%, p < 0.001, respectively). Differentiation between group B and C was marginally significant (p = 0.053).ConclusionThe 8th AJCC/UICC TNM staging system did not clearly distinguish the prognosis of stage II from that of other stages. Including the T2N0-1 group in stage II may improve prognostic stratification.

Current and future techniques for human papilloma virus (HPV) testing in oropharyngeal squamous cell carcinoma.

Despite a reduction in smoking and alcohol consumption, the incidence of oropharyngeal squamous cell carcinoma (OPSCC) is rising. This is attributed to human papilloma virus (HPV) infection and screening for HPV is now recommended in all cases of OPSCC. Despite a variety of clinically available tests and new non-invasive test strategies there is no consensus on which technique is best. This review reports on current techniques for HPV detection in OPSCC and the clinical applicability of emerging techniques. Literature searches of Medline, Embase and clinicaltrials.gov using the search terms 'head and neck neoplasms', 'squamous cell carcinoma' and 'HPV testing' were performed. 45 studies were identified and included. p16 immunohistochemistry (IHC), HPV DNA in situ hybridization (ISH) and HPV polymerase chain reaction (PCR) are the commonest tests to determine HPV status. p16 IHC and HPV DNA PCR are highly sensitive whilst HPV DNA ISH is more specific, these techniques conventionally utilize surgical biopsies. New tests using PCR to screen fine needle aspirates, saliva, brush cytology and serum for HPV are promising but have variable sensitivity and specificity. These non-invasive samples avoid the morbidity of surgical biopsies and need for tissue blocks; their clinical role in screening and surveillance remains largely untested. Further work is needed to validate these tests and define their role.

Genital Human Papillomavirus Infection in Indian HIV-Seropositive Men Who Have Sex With Men.

The incidence of penile cancer in Indian men is high. Little is known about genital human papillomavirus (HPV) infection in Indian HIV-seropositive men who have sex with men (MSM), a population that may be at particularly high risk for genital HPV infection and, potentially, penile cancer. In this study, we assessed the prevalence and risk factors for genital HPV infection in this population. Three hundred HIV-seropositive MSM were recruited from 2 clinical sites in India. They were tested for genital HPV infection using L1 HPV DNA polymerase chain reaction with probes specific for 29 types and a mixture of 10 additional types. Participants received an interviewer-administered questionnaire that included questions on demographics and behaviors. Human papillomavirus data were available from 299 participants. The prevalence of any HPV type in the penis and scrotum was 55% and 54%, respectively. Human papillomavirus type 35 was the most common oncogenic HPV type followed by HPV-16. In multivariate analysis, being the insertive partner with 100+ male partners increased the odds of any penile HPV infection compared with not being insertive with any partners (odds ratio, 2.5; 95% confidence interval, 1.3-5.1). Circumcision was protective against penile HPV infection (odds ratio, 0.39; 95% confidence interval, 0.19-0.76). The prevalence of penile and scrotal HPV infection was high among Indian HIV-seropositive MSM. The most common oncogenic HPV type in this population, HPV-35, is not included in any currently available HPV vaccines. Insertive anal sex with men and lack of circumcision were the primary risk factors for penile HPV infection in this population.

Assessment of HPV screening methods and sample storage in oral lichen planus lesions

Aim:We investigated the human papillomavirus (HPV) DNA distribution in oral lichen planus (OLP) samples using non-invasive methods, with the potential for much broader population screening.Methods:Three different non-invasive collection methods (dry storage, lysis buffer preservation, and use of a commercial tool to rake epithelial cells and immersion of the collection device into the provided rinse solution) were evaluated. HPV DNA was detected in brushed or scraped samples of the lesion site using a commercial HPV DNA polymerase chain reaction kit.Results:In the group of patients with OLP lesions, HPV was detected significantly more frequently than in the healthy control group (lysis buffer for control = 2.1%, p = 0.046; DNA•SAL™ rinse solution for control = 4.2%, p = 0.0003; dry storage for control = 6.3%, p = 0.0001) regardless of the different collection and preservation methods. HPV DNA was detected in 12.85% of the samples preserved in lysis buffer. HPV DNA was detected in 34.28% (p = 0.0048 vs. ly...

Keratinizing odontogenic cysts with a spectrum of verrucoid morphology: investigation of a potential role of human papillomavirus

The role of human papillomavirus (HPV) in keratinizing odontogenic cysts (OC) has only rarely been studied. We describe the clinicopathologic findings in a series of OCs that had unusual keratinization patterns and were investigated for a possible HPV etiology. Tissue samples from 29 patients with keratinizing OCs were studied for light microscopic features suggestive of HPV infection and by an HPV DNA polymerase chain reaction assay. The mean age at presentation was 31.1years; 79.3% of the OCs occurred in the mandible and 46.4% were associated with an impacted tooth. The phenotypic characteristics koilocytes, hypergranulosis, and a verrucous pattern of the cyst-lining epithelium were observed in 69%, 62.1%, and 17.2% of cases, respectively. These histomorphologic features did not, however, correlate with HPV infection. HPV does not appear to play a role in keratinizing OCs and is not responsible for the wart-like histomorphologic features that may be seen in these lesions.

English

Human immune deficiency virus (HIV) infection is associated with a higher risk of human papillomavirus (HPV) positivity and cervical intraepithelial neoplasia (CIN) which may progress to cancer. Hence, it is important to screen all HIV positive women for this cancer which is amenable to cure in early stages. A cross sectional case-control study was conducted on 100 HIV seropositive women and matched seronegative controls attending Lok Nayak Hospital, New Delhi to study the prevalence of HPV infection and abnormal cervical cytopathology. A detailed cervical evaluation including a per-speculum examination, pap smear, cervical scrape for HPV DNA polymerase chain reaction (PCR), visual inspection of cervix with acetic acid, colposcopy and guided biopsy were done. Abnormal pap smears were found in 63% women of which 10% were squamous intraepithelial lesions. Biopsy revealed cervical intraepithelial neoplasia in 2% of all cases (5.6% of all patients biopsied). Prevalence of HPV in HIV positives was 24% compared to 4% in controls (p value 0.001), also bearing a significant correlation with CIN, thus placing HIV positives at a higher risk for cervical dysplasias and cancer. The sensitivity of HPV DNA test for detection of cervical dysplasias was 86.3% and specificity 64.2%. HIV positivity predisposes to invasive cervical cancer on account of immunosuppression and co-existing HPV infection, thereby the need for aggressive screening for cervical intraepithelial lesions. Both HPV DNA PCR and pap smear are optimal screening tools in these women. Key words: Human immune deficiency virus (HIV), cervical human papillomavirus (HPV) infection, cervical intraepithelial neoplasia, pap smear, human papillomavirus (HPV) DNA polymerase chain reaction (PCR).

Abnormal Pap Tests and Human Papillomavirus Infections Among HIV-Infected and Uninfected Women Who Have Sex With Women

To estimate the frequency of abnormal Pap and human papillomavirus (HPV) positivity among human immunodeficiency virus (HIV)-seropositive and -seronegative women who have sex with women (WSW). Pap and HPV DNA polymerase chain reaction tests were obtained every 6 months from women in a US cohort of HIV-seropositive and -seronegative women. Women who have sex with women were women reporting no male and at least 1 female sex partner for 5 years. They were frequency matched 1:5 to women reporting sex only with men (WSM) and assessed using multivariable generalized estimating equation logistic regression models. Paps at study entry were abnormal in 12 (21%) of 49 HIV-seropositive WSW, 151 (64%) of 245 HIV-seropositive WSM, 3 (9%) of 24 HIV-seronegative WSW, and 16 (11%) of 120 HIV-seronegative WSM. Human papillomavirus was found at entry in 18 (42%) HIV-seropositive WSW, 109 (52%) HIV-seropositive WSM, 6 (27%) HIV-seronegative WSW, and 13 (13%) HIV-seronegative WSM. After controlling for HIV serostatus and CD4 count, WSW had marginally lower odds than WSM of Pap abnormality (odds ratio = 0.59, 95% confidence interval = 0.33-1.03) and of HPV (odds ratio = 0.53, 95% confidence interval = 0.32-0.89). After controlling for partner's gender, HIV seropositivity and lower CD4 count were associated with any HPV, oncogenic HPV, any abnormal Pap result, and high-grade squamous intraepithelial lesion or worse (p < .0001 for all). Although risks for abnormal Pap and HPV are modestly lower in WSW than in WSM, both are common in HIV-seropositive women regardless of sexual preference. Both WSW and WSM should be screened similarly.

Verrucous carcinomas of the head and neck, including those with associated squamous cell carcinoma, lack transcriptionally active high-risk human papillomavirus

Most oropharyngeal squamous cell carcinomas (SCC) and histologic variants harbor transcriptionally active human papillomavirus (HPV). While HPV DNA can be found in many non-oropharyngeal head and neck carcinomas, transcriptionally active HPV is rare. Verrucous carcinoma is a variant with bland cytology, warty appearance, locally destructive growth, and lack of metastasis when lacking a frankly invasive carcinoma component. Studies have shown variable rates of HPV DNA and p16 protein expression in such tumors but still have not clearly addressed if the virus has biological activity or clinical relevance in the positive cases. Department files were searched for verrucous neoplasms, including pure verrucous carcinoma, verrucous carcinoma with dysplasia or minimal invasion, and SCC arising in verrucous carcinoma (ie, having a major component of frankly invasive carcinoma). p16 immunohistochemistry, HPV DNA polymerase chain reaction (PCR) and E6/E7 mRNA reverse transcription PCR for high-risk HPV types were performed. Of the 49 cases, 6 (12.2%) showed strong (>50%) staining for p16. HPV DNA was detected in 7/49 (14.3%) cases, but only one case was positive for both p16, and HPV DNA. A total of 36 cases yielded sufficient RNA for RT-PCR (18 verrucous carcinomas, 13 atypical verrucous carcinomas, and 5 SCC arising in verrucous carcinoma). All 36 were negative, including the four p16-positive and three HPV DNA-positive tumors tested. Although a minority of verrucous carcinoma lesions are p16 and HPV DNA positive, transcriptionally active high-risk HPV is uniformly absent. These findings argue that verrucous carcinoma and its related squamous cell carcinomas are not HPV-driven tumors.

Risk factors for anal human papillomavirus infection type 16 among HIV-positive men who have sex with men in San Francisco.

HIV-positive men who have sex with men (MSM) are at high risk of anal cancer compared with the general population. Human papillomavirus (HPV) infection, particularly HPV 16, is causally associated with anal cancer. However, the risk factors for anal HPV 16 infection are poorly understood. We determined the prevalence and risk factors for anal HPV 16 infection in a population of HIV-positive MSM, most of whom were being treated with antiretroviral therapy. Cross-sectional data from the baseline visit of a 4-year prospective cohort study. Three hundred forty-eight HIV-positive MSM were recruited in San Francisco, and they received a detailed sexual behavior risk factor questionnaire. An anal swab was used to collect specimens for HPV type-specific DNA testing using L1 HPV DNA polymerase chain reaction. We used log-binomial multivariable models to determine the risk factors for anal HPV 16 infection. Ninety-two percent of HIV-positive MSM had at least 1 anal HPV type, 80% had at least 1 oncogenic HPV type, and 42% had HPV 16. Non-Hispanic white race and higher level of education were associated with a decreased risk of HPV 16 infection. A higher number of total male partners was associated with HPV 16 (relative risk: 1.6, 95% confidence interval 1.1 to 2.4, P = 0.01) for 201-1000 partners compared with 1-200. Injection drug use was independently associated with anal HPV 16 infection (relative risk: 1.5, 95% confidence interval 1.2 to 1.9, P = 0.003). The prevalence of anal HPV infection, including HPV 16, is high in HIV-positive MSM. HIV-positive MSM should be counseled about the risk associated with increased partners and injection drug use.

Detection of HPV infection in head and neck squamous cell carcinoma: a practical proposal

Detecting human papillomavirus (HPV) infection in head and neck squamous cell carcinoma (HNSCC) is clinically relevant, but there is no agreement about the most appropriate methodology. We have studied 64 oropharyngeal carcinomas using p16 immunohistochemistry, HPV DNA in situ hybridisation (ISH) and HPV DNA polymerase chain reaction (PCR) followed by pyrosequencing. We have also evaluated a new assay, RNAscope, designed to detect HPV E6/E7 RNA transcripts. Using a threshold of 70 % labelled tumour cells, 21 cases (32.8 %) were p16 positive. Of these, 19 cases scored positive with at least one HPV detection assay. Sixteen cases were positive by HPV DNA-ISH, and 18 cases were positive using the E6/E7 RNAscope assay. By PCR and pyrosequencing, HPV16 was detected in 15 cases, while one case each harboured HPV33, 35 and 56. All p16-negative cases were negative using these assays. We conclude that p16 expression is a useful surrogate marker for HPV infection in HNSCC with a high negative predictive value and that p16-positive cases should be further evaluated for HPV infection, preferably by PCR followed by type determination. Using RNase digestion experiments, we show that the RNAscope assay is not suitable for the reliable discrimination between E6/E7 RNA transcripts and viral DNA.

Human papillomavirus prevalence and type in liquid-based cervical samples from Turkish women in a selected risk group

To detect the prevalence and type distribution of human papillomavirus (HPV) in different cytological diagnostic categories. Materials and methods: Between 2007 and 2010, a total of 1014 liquid-based thin preparations of cervical smears were selected and classified according to cytology results. HPV DNA polymerase chain reaction (PCR) was also performed using these samples. HPV DNA-positive samples were genotyped by DNA sequencing. Results: Of those enrolled in the study, 45.3% were negative cytologically, 36.4% had atypical squamous cells of undetermined significance, 0.3% had atypical squamous cells preventing the exclusion of a high-grade squamous intraepithelial lesion, 16.8% showed a low-grade squamous intraepithelial lesion, and 1.3% had an high-grade squamous intraepithelial lesion. Conclusion: PCR assays showed HPV positivity in 63.0% of cytologically negative and 90.8% of cytologically positive samples. The most common types of HPV detected were 16, 6, 18, 31, 66, 56, 53, 81, 45, and 62. Of HPV DNA-positive samples, 47.7% were high, 4.7% were intermediate, and 17.9% were low risk. The high-risk types of HPV detected were 16, 18, 31, 56, 53, 45, 62, 58, 59, 67, 51, 35, 73, 52, 33, 39, 68, and 82.