High-risk Human Papillomavirus Types Research Articles

Abstract 3948: HPV45 and HPV52 prevalence, within-type variants, and precancer/cancer risks differ by race/ethnicity

Abstract Introduction: Persistent infection with a high-risk HPV (HR-HPV) type causes cervical cancer and many other cancers in both men and women. Our published data have shown that the distribution of specific HPV16/35 sublineages and genetic variants differ around the world and confer greater risk of cervical precancer/cancer in the populations where the virus is thought to have originated and coevolved. More studies are needed to evaluate the relationship between race/ethnicity, HR-HPV genetic variation, and precancer/cancer risk. Objective: To evaluate HPV45 and HPV52 genetic variation, and how it relates to a woman’s race/ethnicity and cervical precancer/cancer risk, using HPV whole-genome sequencing of 774 HPV45+ and 719 HPV52+ women in the NCI-Kaiser Permanente Northern California (KPNC) HPV Persistence and Progression cohort; and compared to HPV16/35. Methods: We viral genome sequenced 557 HPV45+ and 345 HPV52+ controls (women with benign infections [≤CIN1]), and 94 HPV45+ and 227 HPV52+ precancer/cancer cases (CIN3+). We evaluated HPV45 and HPV52 prevalence for each type by a woman’s self-reported race/ethnicity, and compared them to HPV16 and HPV35 using existing comparable NCI-KPNC data. To assess associations between HPV45 and HPV52 genetic variation (viral sublineages and SNPs) and infection outcome, we used Fisher’s Exact Tests and logistic regression to estimate the OR and 95% CI. Results: For HPV45, we determined that sublineages have varying histology-specific precancer/cancer risk (ORs ranged from 2-14), and that the sublineage distribution significantly varied by race/ethnicity (p < 0.001). By race, the HPV45 A2 sublineage was most prevalent in Asian/PI women and was associated with the highest risk of CIN3+ in these women compared to other races/ethnicities (OR=5.7, 95% CI=1.5-29.4), while the B2 sublineage was associated with increased risk of CIN3+ in Black women only (OR=6.1, 95% CI=1.2-33.6). The prevalence of each HR-HPV type (HPV16/35/45/52) significantly varied by race/ethnicity (p < 0.001). For each type, White was the most reported race/ethnicity; however, Black women were a greater proportion of the HPV35+ cases compared to other types (15% vs 8-10%), and Asian/PIs were a greater proportion of HPV52+ cases (26% vs 10-13%). HPV52 genome analyses are still underway; we will present the complete study results (i.e., sublineage risk by race) at AACR. Conclusions: We conducted the largest studies of HPV45 and HPV52 genome variation to date. We demonstrate that specific HPV45 sublineages vary by race and are associated with an increased risk of precancer/cancer for specific races/ethnicities, and that HPV52 infection has a greater prevalence in Asian/PI women compared to other racial groups. Our data suggest that certain HR-HPV types and specific sublineages of these types are more common in specific racial/ethnic groups and potentially linked to differing risks by race. Citation Format: Aimee J. Koestler, Chase W. Nelson, Meredith Yeager, Zigui Chen, Laurie Burdett, Sambit K. Mishra, Michael Dean, Elizabeth Suh-Burgmann, Thomas Lorey, Phillip E. Castle, Mark Schiffman, Lisa Mirabello. HPV45 and HPV52 prevalence, within-type variants, and precancer/cancer risks differ by race/ethnicity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3948.

Abstract 4804: Prevalence of anal HPV infection in women attending a gynecology and colposcopy clinic: Assessing the potential impact of HPV vaccination

Abstract Human papillomavirus (HPV) accounts for over 80% of anal cancer cases. In the effort to prevent HPV-related cancers, the 9-valent HPV vaccine emerged as a crucial intervention, targeting HPV types 6, 11, 16, 18, 31, 33, 45, 52, and 58; the previous 4-valent (types 6,11,16,18) and 2-valent (types 16, 18). This study involved 213 women aged 21-50 attending gynecology and colonoscopy clinics in San Juan, Puerto Rico. The study assessed anal HPV prevalence and analyzed vaccination status-related differences. Anal swabs were collected for HPV testing using the ATILA AmpFire, targeting 17 HPV types. This method identified 15 high-risk HPV types (16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, and 68) and two low-risk HPV types (6 and 11). Prevalence of any HPV, HR-HPV, and LR-HPV, and of specific HPV types were calculated by overall and vaccination status. Differences were assessed for statistical significance (p<0.05) using Fisher's exact or Pearson χ2 test. The mean age of women was 36.0 ± 5.9 years. Overall, 19.7% (n=44) had received the HPV vaccine; among them, 52.2% (n=23) had received the 4-valent vaccine, 2.3% had received the 2-valent vaccine, 2.3% had received the 9-valent vaccine, and 43.2% were uncertain about their specific HPV vaccine type. The prevalence of anal HPV infection was 68.5% for any HPV, 61.5% for HR-HPV, and 15.0% for LR-HPV. The most prevalent HR-HPV types were HPV 56 (13.2%), HPV 68 (11.7%), and HPV 66 (11.2%). There were no statistically significant differences in the prevalence of any HPV, HR-HPV, and LR-HPV between vaccinated and unvaccinated women. The most common HR-HPV type among vaccinated were HPV 66 (20.5%), HPV 31 (18.2%), HPV 56 (15.9%), while the most common among unvaccinated women were HPV 56 (12.4%), HPV 68 (12.4%), and HPV 16 (12.4%). Additionally, when evaluating as a grouped variable, the HPV genotypes covered by the 4-valent, unvaccinated women had a significantly higher prevalence of these types as compared to those vaccinated (28.9% vs. 13.6%, respectively, p-value =.03). The unvaccinated women showed a lower prevalence for HPV types covered by the 9-valent vaccine compared to the vaccinated cohort (43.2% vs. 45.5%). However, this difference was not statistically significant. We found a high prevalence of anal HPV among these women, with lower prevalence of 4-valent and 2-valent HPV types among vaccinated women, evidencing the pivotal role of HPV vaccination. Future research should assess the 9-valent vaccine's long-term effectiveness against more HPV types. Funding: Research Centers in Minority Institutions (RCMI), University of Puerto Rico (NIMHD # 2U54MD007600-36), and CAPAC Research Training Program (NCI #R25CA240120). Citation Format: Anelisse Dominicci-Maura, Ana Patricia Ortiz, Josefina Romaguera, Filipa Godoy-Vitorino. Prevalence of anal HPV infection in women attending a gynecology and colposcopy clinic: Assessing the potential impact of HPV vaccination [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4804.

CLIA Validation of a New High Performance HPV ctDNA Assay Covering Multiple High Risk HPV Types

CLIA Validation of a New High Performance HPV ctDNA Assay Covering Multiple High Risk HPV Types

Prevalence of Human Papillomavirus (HPV) and HPV Type Distribution in Penile Samples in Young Men in Denmark: Results 10 Years After Implementation of a Girls-Only HPV Vaccination Program.

In Denmark, a girls-only human papillomavirus (HPV) vaccination program was initiated in 2008-2009. The study aim was to assess the HPV prevalence and type distribution in younger men prior to HPV vaccination in men. The study population was younger men who attended information days regarding military service. At random days (2019-2020), 280 men were included. We collected questionnaire data regarding risk factors for HPV infection and a penile swab for HPV testing. We compared results in this study with those from a previous study of young men (2006-2007). The majority of participants (94%) were 18-20 years old. The median number of lifetime sexual partners was 4. Altogether, 130 men (46.4%) were HPV positive. No infections with HPV types 6, 11, 16, 18, 31, and 45 were detected. The most frequent type was HPV-51 (detected in 11.1%). Comparison showed that the odds of high-risk HPV type infection were higher in 2019-2020 (prevalence odds ratio [POR], 1.7 [95% confidence interval {CI}, 1.1-2.7]) compared with 2006-2007. In contrast, the odds were lower (POR, 0.3 [95% CI, .1-.6]) for HPV types targeted by the 9-valent HPV vaccine. The multicohort girls-only vaccination program has to a large degree protected young men against the HPV types included in the licensed vaccines. This does not speak against gender-neutral vaccination as the HPV prevalence is still high, although consisting largely of less carcinogenic HPV types.

A multiplex RPA-CRISPR/Cas12a-based POCT technique and its application in human papillomavirus (HPV) typing assay

Persistent infection with high-risk human papillomavirus (HR-HPV) is the primary and initiating factor for cervical cancer. With over 200 identified HPV types, including 14 high-risk types that integrate into the host cervical epithelial cell DNA, early determination of HPV infection type is crucial for effective risk stratification and management. Presently, on-site immediate testing during the HPV screening stage, known as Point of Care Testing (POCT), remains immature, severely limiting the scope and scenarios of HPV screening. This study, guided by the genomic sequence patterns of HPV, established a multiplex recombinase polymerase amplification (RPA) technology based on the concept of “universal primers.” This approach achieved the multiple amplification of RPA, coupled with the CRISPR/Cas12a system serving as a medium for signal amplification and conversion. The study successfully constructed a POCT combined detection system, denoted as H-MRC12a (HPV—Multiple RPA—CRISPR/Cas12a), and applied it to high-risk HPV typing detection. The system accomplished the typing detection of six high-risk HPV types (16, 18, 31, 33, 35, and 45) can be completed within 40 min, and the entire process, from sample loading to result interpretation, can be accomplished within 45 min, with a detection depth reaching 1 copy/μL for each high-risk type. Validation of the H-MRC12a detection system’s reproducibility and specificity was further conducted through QPCR on 34 clinical samples. Additionally, this study explored and optimized the multiplex RPA amplification system and CRISPR system at the molecular mechanism level. Furthermore, the primer design strategy developed in this study offers the potential to enhance the throughput of H-MRC12a detection while ensuring sensitivity, providing a novel research avenue for high-throughput detection in Point-of-Care molecular pathogen studies.

Prevalence and Correlation between High-Risk HPV Genotypes and Pap Smear Findings in Bahrain: A Retrospective Approach

Background: Human papillomavirus (HPV) is currently the most common pathogen responsible for cervical cancer (CC), a major cause of cancer-related deaths in women. This study aimed to estimate the current HPV prevalence in Bahrain and to determine the association of high-risk HPV (HR-HPV) genotypes with cytological findings, age, and clinical history of the patients. Methods and Results: Our study used a retrospective approach. Convenience sampling was used to gather 100 cases of HPV-positive women with abnormal and normal Pap smear findings from January 2017 to April 2023 in the Cytology Department at King Hamad University Hospital. Out of 100 HR-HPV positive cases, non-16/18/45 HR-HPV was found in 62%, and 73% had abnormal Pap smear findings. Among Pap smear findings, ASC-US/ASC-H (P=0.038) and LSIL/HSIL(P=0.017) were significantly associated with HR-HPV genotypes.ASC-US was found to be more frequently associated with HPV16+non-16/18/45 HR-HRV and LSIL with HPV18/45+non-16/18/45 HR-HRV. Most HR-HPV cases (59%) were aged ≤40 years, 25% - from 41 to 50 years old, while only 16% were >50. The age group ≤40 had the highest peak with non-16/18/45 HR-HPV genotype (35%) and HPV16 (19%). The predominant genotypes for age groups 41-50 and >50 were non-16/18/45 HR-HPV types (19% and 8%, respectively). Conclusion: In Bahrain, non-16/18/45 HR-HPV infection is becoming more prevalent, with ASC-US being the most common abnormal Pap finding and the highest HPV infection in women aged 40 years or younger.Based on our findings, we recommend effective screening and vaccine programs for women aged 40 years and younger, as early detection can lower infection rates and improve recovery.

Correlation between high-risk HPV infection and p16/Ki-67 abnormalities in Pap samples in a South Eastern Europe cohort.

Cervical cancer (CC) is the fourth most common cause of cancer-related deaths amongst women worldwide. CC represents a major global healthcare issue, and Romania ranks the worst in mortality rates amongst EU countries. However, the early detection of CC can be lifesaving. To understand the testing process undergone by women in Romania, we performed a retrospective study, and investigated a cohort of 83 785 cervical cases from Romanian women aged 15-70, obtained in private-based opportunistic screening. We examined the correlation between Pap smear results, human papilloma virus (HPV) genotyping, and the expression of cell cycle markers p16 and Ki-67. Analysis of Pap results revealed approximately 10% abnormal cases, of which high-grade squamous intraepithelial lesionsconstituted 4.9%. HPV genotyping of 12 185 cases with available Pap results unveiled a range of high-risk HPV (hrHPV) types associated with cervical abnormalities. Notably, 26% of hrHPV-positive cases showed no observable abnormalities. In a subset of cases with abnormal Pap and a type of hrHPV, P16/Ki-67 double-staining was also positive. This study suggests the importance of an integrated diagnostic algorithm that should consider the HPV genotype, Pap smear, and p16/Ki-67 staining. This algorithm should enhance the CC screening accuracy and its management strategies, particularly in those regions with a high disease burden, such as Romania.

The expression analysis of human endogenous retrovirus-K Env, Np9, and Rec transcripts in cervical cancer.

While infection with high-risk human papillomavirus (HPV) types is necessary for cervical cancer (CC) development, it is not enough, and other risk factors are required. Several studies have reported the activation of HERV-K in different cancers; however, the investigation of HERV-K expression levels in CC is scarce. In this study, it was hypothesized that activation of HERV-K could play an essential role in CC development. In this order, the expression levels of HERV-K Env, Np9, and Rec transcripts were investigated on 147 normal to CC uterine cervical tissues using quantitative real-time PCR. The significantly higher levels of HERV-K Env and Np9 transcripts were found in patients with cervical intraepithelial neoplasia (CIN) II-III and CC groups compared to those in the normal/CIN I group. Expression of Rec transcript was also higher only in the CC group than normal/CIN I group. Among CC patients, meaningfully higher levels of HERV-K Env and Np9 transcripts were found in patients with squamous cell carcinoma rather than in adenocarcinoma. When only the HPV 16 positive samples were investigated, it was found that the mean difference in Env and Np9 mRNA levels was meaningfully higher among precancer lesions and the cancer group in comparison with the normal group. However, the Rec mRNA level showed no significant differences. The association between the expression of HERV-K genes was investigated, and a significant positive correlation of Env expression with Np9 transcript was found only in the group with precancer lesions (R = 0.6, p = 0.0037). Moreover, a significant positive correlation was found between Rec and Np9 transcripts in patients with normal cervix tissues (R = 0.26, p = 0.033). However, no correlations were observed between the expression of Env and Rec in the three groups. In conclusion, our results showed that HERV-K transcripts, especially Env and Np9, upregulated during cervical lesion progression. These findings highlight the potential use of HERV-K Env and Np9 as biomarkers for CC diagnosis and prognosis. Further investigation is needed to determine the clinical utility of these markers and whether targeting HERV-K oncogenes could be a viable therapeutic strategy for CC.

Consistency in human papillomavirus type detection between self-collected vaginal specimens and physician-sampled cervical specimens.

With the rising need for accessible cervical cancer screening, self-sampling methods offer a promising alternative to traditional physician-led sampling. This study aims to evaluate the efficacy of the HygeiaTouch Self Sampling Kit for Women in detecting human papillomavirus (HPV) types and predicting cervical lesions. We studied the concordance in identifying high-risk HPV (hrHPV) types between samples collected by physicians and those self-collected by women using a self-sampling kit for validation. Women aged 21-65, fitting into specific categories based on their cervical health history were eligible. Cohen's kappa coefficient to gauge concordance between the two specimen types and relative accuracy metrics in identifying cervical intraepithelial neoplasia (CIN) were also calculated, with physician-sampled specimens serving as a reference. A total of 1210 participants from three institutes were involved. The self-sampling kit closely matched the physician-led method in terms of collecting valid specimens (100% vs. 100%), identifying hrHPV types (kappa: 0.75, 95% confidence interval [95% CI]: 0.72-0.79; agreement: 87.7%, 95% CI: 85.8-89.6) and predicting CIN grade 2 or worse (CIN2+) (relative sensitivity: 0.949, relative accuracy: 0.959). Kappa values varied between 0.71 and 0.83 for different hrHPV types and combinations, with an overall value 0.75 (95% CI: 0.72-0.79) signifying robust compatibility between the two methods. Our study underscores the potential of the HygeiaTouch Self Sampling Kit as a reliable, efficient, and user-friendly alternative to traditional sampling methods. This suggests that self-sampling could be pivotal in expanding cervical cancer screening accessibility and enhancing detection rates.

Human Papillomavirus (HPV) Infection and Risk Behavior in Vaccinated and Non-Vaccinated Paraguayan Young Women.

Cervical cancer is a global health concern and ranks fourth among the most prevalent cancers in women worldwide. Human papillomavirus (HPV) infection is a known precursor of cervical cancer and preventive measures include prophylactic vaccines. This study focused on sexually active Paraguayan women aged 18-25 years, exploring the intersection of HPV vaccination and sexual behavior. Among 254 participants, 40.9% received the Gardasil-4 vaccine, with no significant differences in sexual behavior between the vaccinated and unvaccinated sexually active groups. However, a notable decrease in the prevalence of HPV among the vaccinated women highlights the efficacy of this vaccine in reducing infections. The prevalence of any HPV type was 37.5% in vaccinated participants compared to 56.7% in unvaccinated participants (p = 0.0026). High-risk HPV types showed a significant difference, with a prevalence of 26.0% in vaccinated women compared with 52.7% in unvaccinated women (p < 0.001). Although a potential decline in genital warts was observed among the vaccinated individuals, statistical significance (p = 0.0564) was not reached. Despite the challenges in achieving high vaccination coverage, the observed reduction in HPV prevalence underscores the importance of ongoing monitoring, healthcare professional recommendations, and comprehensive risk management. These findings contribute to dispelling concerns about HPV vaccination influencing sexual behavior, advocating further large-scale research to explore the impact of vaccines on various HPV types and potential cross-protection.

Analytical evaluation of the automated genotyping system (GenPlex) compared to a traditional real-time PCR assay for the detection of high-risk human papillomaviruses.

The detection of high-risk human papillomaviruses (HPVs) is crucial for early screening and preventing cervical cancer. However, the substantial workload in high-level hospitals or the limited resources in primary-level hospitals hinder widespread testing. To address this issue, we explored a sample-to-answer genotyping system and assessed its performance by comparing it with the traditional real-time polymerase chain reaction(PCR) method conducted manually. Samples randomly selected from those undergoing routine real-time PCR detection were re-analyzed using the fully automatic GenPlex® system. This system identifies 24 types of HPV through a combination of ordinary PCR and microarray-based reverse hybridization. Inconsistent results were confirmed by repeated testing with both methods, and the κ concordance test was employed to evaluate differences between the two methods. A total of 365 samples were randomly selected from 7259 women. According to real-time PCR results, 76 were high-risk HPV negative, and 289 were positive. The GenPlex® system achieved a κ value greater than 0.9 (ranging from 0.920 to 1.000, p < 0.0001) for 14 types of high-risk HPV, except HPV 51 (κ = 0.697, p < 0.0001). However, the inconsistent results in high-risk HPV 51 were revealed to be false positive in real-time PCR by other method. When counting by samples without discriminating the high-risk HPV type, the results of both methods were entirely consistent (κ = 1.000, p < 0.0001). Notably, the GenPlex® system identified more positive cases, with 73 having anHPV type not covered by real-time PCR, and 20 potentially due to low DNA concentration undetectable by the latter. Compared with the routinely used real-time PCR assay, the GenPlex® system demonstrated high consistency. Importantly, the system's advantages in automatic operation and a sealed lab-on-chip format respectively reduce manual work and prevent aerosol pollution. For widespread use of GenPlex® system, formal clinical validation following international criteria should be warranted.

A population based study on human papillomavirus infection and associated risk factors among women of the remote South Andaman Island, India

BackgroundHuman papillomavirus (HPV) is associated with cervical cancer and cervical dysplasia worldwide. Data on HPV prevalence in a region is important because it serves as a predictor of the likelihood of the population in that particular region acquiring cervical cancer. Moreover, with the availability of effective vaccines, the public health system must be aware of the preponderance of HPV to implement the vaccine. The present study was designed to understand the prevalence of HPV and associated factors among the women of South Andaman Island.MethodsA cross-sectional study was conducted among married women of reproductive age (18–59 years) from South Andaman District from 2018 to 2022. Cervical scrapes were collected from participants after obtaining informed written consent for HPV molecular testing (HPV DNA) such as PCR assay. Demographic data was collected using a standard questionnaire and statistical analyses were performed to determine the associated factors.ResultsThe study showed prevalence of HPV as 5.9%(95% CI: 3.9–7.9) and prevalence of HR-HPV16 was 4.1% (95% CI 2.6 – 5.5) and HR-HPV18 prevalence was 1.8(95% CI: 0.6–3). The independent factors associated the HPV positivity were age above 55 years, menopause, post-menopausal bleeding, blood-stained vaginal discharge and loss of weight. Age was associated with all HPV infections among the South Andaman women.ConclusionsHPV 16 was reported as the predominant high risk HPV type circulating among women of South Andaman. Cervical cancer and precancerous lesions were significantly associated with HPV positivity and High risk HPV 16. Based on the knowledge of the risk factors associated with HPV, implementation of stronger public health awareness and prophylactic HPV vaccination is crucial among the women of this remote island.

Downregulation of LAMB3 Altered the Carcinogenic Properties of Human Papillomavirus 16-Positive Cervical Cancer Cells.

Nearly all cervical cancer cases are caused by infection with high-risk human papillomavirus (HR-HPV) types. The mechanism of cervical cell transformation is related to the powerful action of viral oncoproteins and cellular gene alterations. Transcriptomic data from cervical cancer and normal cervical cells were utilized to identify upregulated genes and their associated pathways. The laminin subunit beta-3 (LAMB3) mRNAwas overexpressed in cervical cancer and was chosen for functional analysis. The LAMB3 was predominantly expressed in the extracellular region and the plasma membrane, which play a role in protein binding and cell adhesion molecule binding, leading to cell migration and tissue development. LAMB3 was found to be implicated in the pathway in cancer and the PI3K-AKT signaling pathway. LAMB3 knockdown decreased cell migration, invasion, anchorage-dependent and anchorage-independent cell growth and increased the number of apoptotic cells. These effects were linked to a decrease in protein levels involved in the PI3K-AKT signaling pathway and an increase in p53 protein. This study demonstrated that LAMB3 could promote cervical cancer cell migration, invasion and survival.

Enhancing Cervical Cancer Screening: Review of p16/Ki-67 Dual Staining as a Promising Triage Strategy.

Cervical cancer, primarily caused by high-risk human papillomavirus (HR-HPV) types 16 and 18, is a major global health concern. Persistent HR-HPV infection can progress from reversible precancerous lesions to invasive cervical cancer, which is driven by the oncogenic activity of human papillomavirus (HPV) genes, particularly E6 and E7. Traditional screening methods, including cytology and HPV testing, have limited sensitivity and specificity. This review explores the application of p16/Ki-67 dual-staining cytology for cervical cancer screening. This advanced immunocytochemical method allows for simultaneously detecting p16 and Ki-67 proteins within cervical epithelial cells, offering a more specific approach for triaging HPV-positive women. Dual staining and traditional methods are compared, demonstrating their high sensitivity and negative predictive value but low specificity. The increased sensitivity of dual staining results in higher detection rates of CIN2+ lesions, which is crucial for preventing cervical cancer progression. However, its low specificity may lead to increased false-positive results and unnecessary biopsies. The implications of integrating dual staining into contemporary screening strategies, particularly considering the evolving landscape of HPV vaccination and changes in HPV genotype prevalence, are also discussed. New guidelines and further research are necessary to elucidate the long-term effects of integrating dual staining into screening protocols.

Efficacy and immunogenicity of AS04-HPV-16/18 vaccine in females with existing cervical HR-HPV infection at first vaccination: A pooled analysis of four large clinical trials worldwide.

Females with existing high-risk HPV (HR-HPV) infections remain at risk of subsequent multiple or recurrent infections, on which benefit from HPV vaccines was under-reported. We pooled individual-level data from four large-scale, RCTs of AS04-HPV-16/18 vaccine to evaluate efficacy and immunogenicity in females DNA-positive to any HR-HPV types at first vaccination. Females receiving the AS04-HPV-16/18 vaccine in the original RCTs constituted the vaccine group in the present study, while those unvaccinated served as the control group. Vaccine efficacy (VE) against new infections and associated cervical intraepithelial neoplasia (CIN) 2+ in females DNA-negative to the considered HR-HPV type but positive to any other HR-HPV types, VE against reinfections in females DNA-positive to the considered HR-HPV type but cleared naturally during later follow-up, and levels of anti-HPV-16/18 IgG were assessed. Our final analyses included 5137 females (vaccine group = 2532, control group = 2605). The median follow-up time was 47.88 months (IQR: 45.72-50.04). For the prevention of precancerous lesions related to the non-infected HR-HPV types at baseline, VE against HPV-16/18 related CIN 2+ was 82.70% (95% CI: 63.70-93.00%). For the prevention of reinfections related to the infected HR-HPV types following natural clearance, VE against HPV-16/18 12MPI was non-significant (p > .05), albeit robust immunity persisted for at least 48 months. Females with existing HR-HPV infections at first vaccination still benefit from vaccination in preventing precancers related to the non-infected types at baseline. VE against reinfections related to the infected types following natural clearance remains to be further investigated.

AmpFire HPV and ScreenFire RS HPV validation trial.

The human papillomavirus (HPV) screening assays from Atila Biosystems, including the new AmpFire (14 type) and ScreenFire RS (13 type), were subjected to a series of validation tests. We used a set of samples from the Chinese Multi-Site Screening Trial (previously tested with cobas 4800 and the next-generation SeqHPV) to satisfy Meijer's criteria for clinical end-point validation. We selected 556 self-collected specimens composed of 273 high-risk HPV (hrHPV) positives and 283 hrHPV negatives on the cobas 4800 and SeqHPV. Of the 273 hrHPV-positive cases, 108 had a disease end point of cervical intraepithelial neoplasia grade 2 (CIN2) or higher, including 47 with cervical intraepithelial neoplasia grade 3 (CIN3+) or higher. We simulated the VALGENT framework for inter- and intralaboratory validation and evaluated the new 4-channel risk-stratified ScreenFire assay in a hierarchal fashion. Both AmpFire and ScreenFire detected 106 (98.1%) of 108 cases with CIN2 or higher, with specificities of 56.7% and 58.1%, respectively. Intralaboratory concordance for 2 runs of AmpFire and ScreenFire was 95.13% and 96.03%, respectively, for overall hrHPV types and 99.10% and 99.46%, respectively, for HPV 16. The interlaboratory concordance of AmpFire and ScreenFire was 93.68% and 94.04% for overall hrHPV and 98.92% and 99.28%, respectively, for HPV 16. Other genotype correlation percentages were similarly high, with κs ranging from 0.86 to 0.94. The ScreenFire RS assay demonstrated excellent "genotype-specific concordance" when evaluated for "clinical guidance" in a hierarchal fashion (noting only the highest risk channel) with both the cobas 4800 and SeqHPV for less than CIN2, CIN2, and CIN3 or higher. The excellent intra- and interlaboratory reproducibility and the established clinical performance, together with the platforms' simplicity, make these assays particularly applicable to low-resource settings.

Human Papillomaviruses (HPVs)—A Review for Diagnosis and Vaccination

Human papillomaviruses (HPVs) are a group of more than 150 related viruses. They are called papillomaviruses because some of them cause papillomas, which are more commonly known as warts. Some types of HPV only grow in skin, while others grow in mucous membranes such as the mouth, throat, or vagina. All types of HPV are spread by contact (touch). More than 40 types of HPV can be passed on through sexual contact. Most sexually active people are infected with one or more of these HPV types at some point in their lives. At least a dozen of these HPV types are known to cause cancer. While HPV infections are very common, cancer caused by HPV is not. Most people infected with HPV will not develop cancer-related to the infection. However, some people with long-lasting infections of high-risk HPV types are at risk of developing cancer. HPV infections of the mucous membranes can cause genital warts, but they usually have no symptoms. There are no effective medicines or other treatments for HPV, other than removing or destroying cells that are known to be infected. But in most people, the body’s immune system controls the HPV infection or gets rid of it over time. To learn more, see HPV and HPV Testing.

CRISPR-Based Multiplex Detection of Human Papillomaviruses for One-Pot Point-of-Care Diagnostics.

The World Health Organization's global initiative toward eliminating high-risk Human Papillomavirus (hrHPV)-related cancers recommends DNA testing over visual inspection in all settings for primary cancer screening and HPV eradication by 2100. However, multiple hrHPV types cause different types of cancers, and there is a pressing need for an easy-to-use, multiplex point-of-care diagnostic platform for detecting different hrHPV types. Recently, CRISPR-Cas systems have been repurposed for point-of-care detection. Here, we established a CRISPR-Cas multiplexed diagnostic assay (CRISPRD) to detect cervical cancer-causing hrHPVs in one reaction (one-pot assay). We harnessed the compatibility of thermostable AapCas12b, TccCas13a, and HheCas13a nucleases with isothermal amplification and successfully detected HPV16 and HPV18, along with an internal control in a single-pot assay with a limit of detection of 10 copies and 100% specificity. This platform offers a rapid and practical solution for the multiplex detection of hrHPVs, which may facilitate large-scale hrHPV point-of-care screening. Furthermore, the CRISPRD platform programmability enables it to be adapted for the multiplex detection of any two nucleic acid biomarkers as well as internal control.

Assessing the feasibility of a multimodal liquid biopsy for the diagnosis of HPV-associated oropharyngeal squamous cell carcinoma.

In this feasibility study, we explored the combined use of circulating tumor human papillomavirus (HPV) DNA (ctHPVDNA) and HPV serology as diagnostic tests for HPV-associated oropharyngeal squamous cell carcinoma (OPSCC). Among patients with research-banked serum or plasma at diagnosis, IgG antibodies to oncoproteins from HPV types 16, 18, 31, 33, 35, 45, 52, and 58 were detected with multiplex serology. Positivity for HPV 16 was defined based on detection of combinations of anti-E6, E1, E2, and E7 and for other high-risk types on detection of anti-E6 and anti-E7. Circulating tumor HPV DNA was detected by custom digital droplet polymerase chain reaction (ddPCR) assays for HPV types 16, 18, 33, 35, and 45. p16 immunohistochemistry and high-risk HPV RNA in situ hybridization (ISH) using a cocktail of 18 high-risk HPV types were performed on tissue. Of 75 patients, 67 (89.3%) were HPV-associated (p16 and HPV RNA ISH positive) and 8 (10.7%) were HPV-independent. All 8 HPV-independent patients were seronegative and negative for ctHPVDNA (100% specificity). Serology was positive in 53 (79.1%) of 67 HPV-associated patients, while ddPCR was positive for ctHPVDNA in 59 (88.6%) of 67 HPV-associated patients. Requiring both tests to be positive resulted in a sensitivity of 50 (74.6%) of 67 while combining assays (either positive) improved sensitivity to 62 (92.6%) of 67. Compared to HPV RNA ISH, HPV serology and ctHPVDNA are sensitive and highly specific biomarkers for HPV-associated OPSCC at the time of presentation.

Feasibility and Acceptability of Self-sampling for Human Papillomavirus DNA Testing Among Asymptomatic Women in Rural Delta State, Nigeria

Background: Self-sampling and human papillomavirus (HPV) DNA testing can be vital tools in cervical cancer screening for hard-to-reach women with limited access to health care in sub-Saharan Africa. This study explored the feasibility and acceptability of self-sampling for HPV testing among asymptomatic women in Orhuwhorun community in Delta State. Methods: This was a cross-sectional study of 230 women aged between 30 - 65years, enrolled by a multi-stage sampling method. Recruited women were asked to provide self-collected vaginal samples between May and June, 2021. HPV detection and genotyping was done using 21 HPV Geno-Array Diagnostic kit. Participants were asked to complete questionnaires after self-sampling to point out their experiences and acceptability of HPV self-sampling. Results: An excellent feasibility of self-sampling (95.2%) was observed. The acceptability rate of self-sampling was 93.0%, and 99.6% (229/230) of the participants were confident that they used the device correctly. The quality of self-sampling was satisfactory in 100% of the samples; 21.1% (48/228) of the samples were positive for HPV, including 12.3% (28/228) with high-risk HPV types,2.6% (6/228) with probable high risk HPV types and 1.8% (4/228)with low-risk HPV types. Multiple HPV infection occurred in 10 cases (4.4%). Conclusion: This study indicates that self-sampling is a feasible and acceptable approach for cervical cancer screening among women in rural Delta State. Thus, the government should consider self-sampling as a valuable strategy in implementing national cancer screening programme.