Human Papillomavirus Type Research Articles

Association of Bacterial Composition and Diversity in the Cervical Tumor Microbiome with HPV Genotype in a Large, International Patient Cohort

Tumor bacterial composition is strongly associated with response to cancer therapy, and is impacted by environment, including geography. Human papillomavirus (HPV) genotypic diversity composition and load are dynamic during pelvic radiation (RT) and correlate with differential responses to RT in cervical cancer patients. In this multi-institutional, collaborative study, we aimed to explore associations between bacterial composition, HPV serotypes, and geographical distribution in an international patient population. Cervical swabs were collected from 287 patients diagnosed with cervical cancer/ dysplasia in four locations: Houston, USA (TX; N = 94), Ethiopia (ETH; N = 85), Puerto Rico (PR; N = 71), and Botswana (BOT; N = 37). Swabs were collected prior to treatment and were subjected to 16S V4 rRNA gene sequencing and HPV genotyping. We compared HPV types and geography via Chi-squared test. We analyzed bacterial composition, alpha diversity (ANOVA), and beta diversity (principal coordinates analysis [PCoA] with PERMANOVA) for HPV type and geography. We used Linear Discriminant Effect Size (LEfSe) analysis to distinguish taxa associated with HPV types. Overall, the global bacterial composition for patients with cancer or dysplasia did not significantly vary by location. However, the proportion of patients with each HPV type varied by location (p<0.01); HPV16 was most frequent in TX (54%), BOT (70%) and ETH (61%), while HPV18 was most frequent in PR (62%). The proportion of patients with HPV low-risk/negative tumors was highest in ETH (25%) compared to other sites (2% - 14%). Patients with HPV 16 had significantly higher bacterial alpha diversity across locations (all p<0.01). The bacterial composition also differed by HPV type across locations (p = 0.01). On LEfSe, bacterial genera enriched in HPV 16 samples were Bacteroides, Clostridium, and Prevotella. Non-HPV16 tumors were enriched in species of Lactobacillus and Gardnerella and HPV 18 and high-risk type tumors were enriched in Escherichia. In thislarge, international cohort of cervical cancer and dysplasia patients, bacterial composition was more closely associated with cervical HPV genotype than with geography. This finding has implications for the development of biomarkers and interventions aimed at improving chemotherapy and radiation response through manipulation of the microbiome.

Clinical Validation of HPV ctDNA for Early Detection of Residual Disease Following Chemoradiation in Cervical Cancer

Despite chemoradiation (CRT), 30-40% of patients with locally advanced cervical cancer relapse. Most cases are caused by human papilloma virus (HPV), and HPV circulating tumor DNA (ctDNA) may identify patients at highest risk of relapse. Our previous pilot study showed that detectable HPV ctDNA at the end of CRT is associated with inferior progression-free survival (PFS) using digital polymerase chain reaction (dPCR), and that a next generation sequencing approach (HPV-seq) may outperform dPCR. We hypothesized that HPV ctDNA may identify cervical cancer patients at increased risk of relapse following CRT and aimed to prospectively validate HPV ctDNA as a tool for early detection of residual disease. This prospective, multicenter validation study accrued 70 patients with HPV+ stage IB-IVA cervical cancer treated with definitive CRT from 2017-2022. Patients underwent phlebotomy at baseline, end of, 4-6 weeks and 3 months post CRT for HPV ctDNA levels. HPV genotyping was performed on the baseline plasma sample using HPV-seq. HPV genotype-specific DNA levels in plasma were quantified using both dPCR and HPV-seq. PFS was estimated using the Kaplan-Meier method and compared using the log rank test. Multivariable Cox regression analyses incorporating stage and HPV ctDNA detectability assessed independent prognostic factors associated with PFS. At the time of abstract, results for 67 patients were available. The majority had squamous histology (84%) and stage IIB (36%) or IIIC1 (25%) disease. HPV genotyping using HPV-seq revealed 54% (36/67) of cases harboring HPV-16, and 46% harboring other HPV types: 15 HPV-18; 5 HPV-59; 2 HPV-31; 2 HPV-33; 2 HPV-52; 1 each HPV-39, HPV-45, HPV-53, HPV-58, and HPV-82. With a median follow up of 2.2 (range 0.4 - 5.2) years, there were 21 PFS events. Most recurrences (14/21) were distant and/or paraaortic; 4 local and nodal/distant; 2 pelvic nodal; and 1 local. Patients with detectable HPV ctDNA on dPCR at the end of, 4-6 weeks and 3 months post CRT had significantly worse 2-year PFS compared to those with undetectable HPV ctDNA (78 vs 52%, p = 0.04; 82 vs 26%, p < 0.001; and 80 vs 23%, p = < 0.001, respectively). HPV-seq showed similar results (87 vs 55%, p = 0.02; 81 vs 45%, p = 0.003; and 84 vs 31%, p = < 0.001, respectively). On multivariable analyses, detectable HPV ctDNA on dPCR and HPV-seq remained independently associated with inferior PFS (see table). HPV-seq enables HPV genotyping directly from plasma in locally advanced cervical cancer. Persistent HPV ctDNA following CRT is independently associated with inferior PFS in this prospective validation study. HPV ctDNA testing can be used to identify, as early as at the end of CRT, patients at high risk of recurrence in future treatment intensification trials.

A rapid HPV typing assay to support global cervical cancer screening and risk-based management: A cross-sectional study.

The World Health Organization recommends human papillomavirus (HPV) testing for cervical screening. Extended genotyping can identify the highest-risk HPV-positive women. An inexpensive, rapid, mobile isothermal amplification assay (ScreenFire HPV RS test) was recently redesigned to yield four channels ordered by cancer risk (ie, hierarchical approach): HPV16, HPV18/45, HPV31/33/35/52/58 and HPV39/51/56/59/68. Stored specimens from 2076 women (mean age 30.9) enrolled in a colposcopy clinic, with high HPV prevalence, were tested with ScreenFire. We calculated hierarchical channel positivity and non-hierarchical channel and type positivity, according to histologic diagnosis (256 cancer, 350 cervical intraepithelial neoplasia [CIN]3, 409 CIN2, 1020 < CIN2) and known virologic reference results (Linear Array and TypeSeq). Additionally, we analyzed ScreenFire time-to-positive up to 60 min by channel and histology. Overall clinical sensitivity for CIN3+ was 94.7% (95% confidence interval 92.6-96.4), similar to Linear Array (92.3, 89.7-94.3) and TypeSeq (96.0, 93.9-97.6). Sensitivity was high for all types and channels. The hierarchical approach was well in line with HPV typing and histologic diagnosis, prioritizing higher risk women having HPV16 and precancer. For HPV16, time-to-positive was shorter in women with precancer. ScreenFire showed excellent agreement with research reference typing tests and detection of CIN2+. Risk-based type results could help guide clinical management of HPV-positive women. Time-to-positive combined with genotyping might be useful. ScreenFire is rapid, mobile, relatively inexpensive and designed for implementation of HPV-based screening and management, including in lower-resource settings. Further validation in screening by self-sampling and practical effectiveness merit evaluation.

HPV Circulating Cell-Free DNA Kinetics in Cervical Cancer Patients Undergoing Definitive Chemoradiation

The human papilloma virus (HPV) is a significant cause of cervical cancer and viral DNA can be detected in the blood of patients with cervical cancer (cfHPV-DNA). We hypothesized that detecting HPV cfDNA before, during and after chemoradiation (CRT) could provide insights into disease extent, clinical staging, and treatment response. Forty-seven patients with cervical cancer were enrolled on this study between 2017 and 2022, either as part of a standard-of-care (SOC) treatment banking protocol (33 patients) or as part of a clinical trial combining a therapeutic HPV vaccine (PDS0101; Immunocerv, 14 patients). Longitudinal plasma samples were collected from each patient as baseline, during week 1, 3 or 5 of CRT. cfHPV-DNA was quantified using droplet digital PCR targeting the HPV E6/E7 oncogenes of 13 high-risk types based on analysis of cervical tumor genotype (AmpFire). Clinical covariates, including FIGO stage, primary tumor size, and treatment response were studied using appropriate statistical tests. All 47 patients had detectable HPV cfDNA during CRT with 38 out of 47 having HPV type 16 detected. The median cfDNA at baseline was 24.5 copies/mL, with a range of 0 to 157,638 copies/mL. Of the 35 patients with at least three measurements, 20 (57%) had peak cfDNA counts at week 3, and 30 out of 35 showed a decline in cfDNA counts at week 5 compared to week 3. The proportion of patients who cleared cfDNA (<16 copies/mL) increased with each week of CRT, reaching 75% at week 5. Baseline cfDNA counts were associated with para-aortic nodal involvement (p<0.0001) but not with FIGO stage or gross tumor volume. A greater proportion of patients treated with therapeutic HPV-directed vaccine had clearance of cfDNA counts as compared to those treated with SOC (at week 3, 38% vs 5%, P = 0.02 and week 5, 79% vs 22%, P = 0.0054) CONCLUSION: HPV cfDNA levels change dynamically throughout definitive CRT and peak during the first 3 weeks for the majority of patients. Treatment with a therapeutic HPV vaccine was associated with a more rapid decline in cfHPV DNA. Further analysis of cfDNA kinetics could provide valuable information on the relationship between cfDNA levels, treatment response, and clinical outcomes.

Genetic variation of E6 and E7 genes of human papillomavirus type 16 from central China

BackgroundPersistent high-risk human papillomavirus (HR-HPV) infection is an important factor in the development of cervical cancer, and human papillomavirus type 16 (HPV-16) is the most common HR-HPV type worldwide. The oncogenic potential of HPV-16 is closely related to viral sequence variation.MethodsIn order to clarify the variant characteristics of HPV-16 E6 and E7 genes in central China, E6 and E7 sequences of 205 HPV‐16 positive samples were amplified by polymerase chain reaction. PCR products of E6 and E7 genes were further sequenced and subjected to variation analysis, phylogenetic analysis, selective pressure analysis and B-cell epitope prediction.ResultsTwenty-six single nucleotide variants were observed in E6 sequence, including 21 non-synonymous and 5 synonymous variants. Twelve single nucleotide variants were identified in E7 sequence, including 6 non-synonymous and 6 synonymous variants. Four new variants were found. Furthermore, nucleotide variation A647G (N29S) in E7 was significantly related to the higher risk of HSIL and cervical cancer. Phylogenetic analysis showed that the E6 and E7 sequences were all distributed in A lineage. No positively selected site was found in HPV-16 E6 and E7 sequences. Non-conservative substitutions in E6, H31Y, D32N, D32E, I34M, L35V, E36Q, L45P, N65S and K75T, affected multiple B-cell epitopes. However, the variation of E7 gene had little impact on the corresponding B-cell epitopes (score < 0.85).ConclusionHPV-16 E6 and E7 sequences variation data may contribute to HR-HPV prevention and vaccine development in Jingzhou, central China.

Female genital schistosomiasis, human papilloma virus infection, and cervical cancer in rural Madagascar: a cross sectional study

BackgroundWomen’s health in resource-limited settings can benefit from the integrated management of high-burden diseases, such as female genital schistosomiasis (FGS) and human papilloma virus (HPV)-related cervical cancer. In schistosomiasis-endemic countries such as Madagascar, data on FGS and HPV prevalence are lacking as well as preventive measures for both conditions. This study aims to estimate the prevalence of FGS and HPV in rural Madagascar, and to examine associated risk factors to identify opportunities for improving women’s health.MethodsAfter initial community outreach activities, interested women aged 18–49 years were recruited consecutively in 2021 at three primary health care centers in the district of Marovoay. FGS was detected by colposcopy. Colposcopy images were double-blind reviewed by two independent specialists. A Luminex bead-based assay was performed on cervical vaginal lavage specimens for HPV typing. Crude (CPR) and adjusted prevalence ratios (APR) of associations between selected factors and FGS and HPV positivity were estimated using univariable and multivariable binary Poisson regression with 95% confidence intervals (CIs).ResultsAmong 500 women enrolled, 302 had complete information on FGS and HPV diagnosis, and were thus eligible for analysis. Within the sample, 189 (62.6%, 95% CI: 56.9–68.1) cases of FGS were detected. A total of 129 women (42.7%, 95% CI: 37.1–48.5) tested positive for HPV. In total, 80 women (26.5%, 95% CI: 21.6–31.8]) tested positive for both conditions. No association was observed between FGS and HPV positivity, while previous pregnancy (APR = 0.65, 95% CI: 0.43–0.78) and older age (APR = 0.59, 95% CI: 0.42–0.81) are showing a negative association with HPV infection compared to no previous pregnancy and younger age groups.ConclusionsThe results of the study show that FGS and HPV are highly prevalent in rural Madagascar. The concurrent prevalence of these two conditions requires urgent adaptations of public health strategies to improve women’s health, such as integrated services at primary level of care.Graphical

Broad-Spectrum Detection of HPV in Male Genital Samples Using Target-Enriched Whole-Genome Sequencing.

Most human papillomavirus (HPV) surveillance studies target 30-50 of the more than 200 known types. We applied our recently described enriched whole-genome sequencing (eWGS) assay to demonstrate the impact of detecting all known and novel HPV types in male genital samples (n = 50). HPV was detected in nearly all (82%) samples, (mean number of types/samples 13.6; range 1-85), and nearly all HPV-positive samples included types in multiple genera (88%). A total of 560 HPV detections (237 unique HPV types: 46 alpha, 55 beta, 135 gamma, and 1 mu types) were made. The most frequently detected HPV types were alpha (HPV90, 43, and 74), beta (HPV115, 195, and 120), and gamma (HPV134, mSD2, and HPV50). High-risk alpha types (HPV16, 18, 31, 39, 52, and 58) were not common. A novel gamma type was identified (now officially HPV229) along with 90 unclassified types. This pilot study demonstrates the utility of the eWGS assay for broad-spectrum type detection and suggests a significantly higher type diversity in males compared to females that warrants further study.

Prevalence and distribution of high-risk Human Papillomavirus in Western Türkiye between 2019 and 2022.

Human Papillomavirus (HPV) is a primarily sexually transmitted human pathogen linked with the formation of anogenital warts and several types of cancer including cervical cancer, oropharyngeal cancer, penile cancer, and anal cancer. HPV genotypes are divided into high-risk and low-risk in accordance with their potential for carcinogenesis. The most notable high-risk genotypes are HPV-16 and HPV-18. In this study we report on the PCR analysis results of the cervical smear results of 328 females, and 50 males who have presented to a tertiary care university hospital in the port city of İzmir, Türkiye between August 2019 and August 2022. HPV-DNA positivity percentage was determined as 25.3% (96/378). 5 of 378 patients were HPV-18 positive (age range 31-80) whereas 27 patients were HPV-16 positive (age range 29-66). 64 patients were positive for other high-risk HPV types (age range 27-56). The positivity for HPV-16, HPV-18 and other high-risk HPV types were determined as 7.14%, 1.3% and 16.9% respectively. Our results demonstrate a 25.3% (96/378) positivity among tested patients moreover, out of 96 positive cases at least 32 are vaccine preventable. This study indicates the need for higher coverage of HPV vaccination in Western Türkiye.

Immunogenicity and safety of an Escherichia coli-produced human papillomavirus (types 6/11/16/18/31/33/45/52/58) L1 virus-like-particle vaccine: a phase 2 double-blind, randomized, controlled trial

The Escherichia coli-produced human papillomavirus (HPV) 16/18 bivalent vaccine (Cecolin) has received prequalification by the World Health Organization based on its high efficacy and good safety profile. We aimed to evaluate the immunogenicity and safety of the second-generation nonavalent HPV 6/11/16/18/31/33/45/52/58 vaccine (Cecolin 9) through the randomized, blinded phase 2 clinical trial. Eligible healthy women aged 18–45 years were randomly (1:1) allocated to receive three doses of 1.0 mL (270 µg) of Cecolin 9 or placebo with a 0-1-6-month schedule. The primary endpoint was the seroconversion rate and geometric mean titer of neutralizing antibodies (nAbs) one month after the full vaccination course (month 7). The secondary endpoint was the safety profile including solicited adverse reactions occurring within 7 d, adverse events (AEs) occurring within 30 d after each dose, and serious adverse events (SAEs) occurring during the 7-month follow-up period. In total, 627 volunteers were enrolled and randomly assigned to Cecolin 9 (n = 313) or placebo (n = 314) group in Jiangsu Province, China. Almost all participants in the per-protocol set for immunogenicity (PPS-I) seroconverted for nAbs against all the nine HPV types at month 7, while two failed to seroconvert for HPV 11 and one did not seroconvert for HPV 52. The incidence rates of total AEs in the Cecolin 9 and placebo groups were 80.8% and 72.9%, respectively, with the majority of them being mild and recovering shortly. None of the SAEs were considered related to vaccination. In conclusion, the E. coli-produced 9-valent HPV (9vHPV) vaccine candidate was well tolerated and immunogenic, which warrants further efficacy studies in larger populations.

The High-Risk Human Papillomavirus Type Influences the Tissue Microenvironment in Cervical Intraepithelial Neoplasia Grade 2.

High-risk, cancer-causing human papillomavirus (HPV) types are associated with cervical precancer and cancer. A high proportion of high-risk HPV precancer lesions undergo immune-mediated regression. The purpose of this study was to determine if the tissue microenvironment of HPV16 and 18 (HPV16/18) cervical intraepithelial neoplasia grade 2 lesions differed from other high-risk types (HPV 'other'). Consistent with other studies, we found that progression to higher-grade disease was more frequent in HPV16/18 lesions when compared with HPV 'other' lesions. HPV16/18 lesions were significantly more likely to be indoleamine 2,3,-dioxygenase 1 (IDO1)-positive and were associated with reduced CD8 and FoxP3 T cells in the lesion. In the stroma, reduced Tbet- and CD32-positive cells and increased Blimp1-positive cells were significantly associated with HPV16/18 lesions when compared with HPV 'other' types. On analysis of the IDO1-positive tissues, lesional IDO1 was associated with significantly decreased numbers of CD4-, CD8-, and FoxP3-positive cells in the stroma compared with IDO1-negative tissues. These data suggest that IDO1 expression may impair infiltration of CD4, CD8, and FoxP3 cells into the stroma beneath the precancer lesion. Increased expression of IDO1 may contribute to immune avoidance and an increased frequency of disease progression in HPV16- and 18-positive lesions.

Human papillomavirus vaccination and anal high-grade precancerous lesions and cancer-a real-world effectiveness study.

Human papillomavirus (HPV) vaccination has shown high efficacy against anal HPV infection and lesions in clinical trials, and the HPV prevalence and type distribution in anal precancers and cancer predict a high preventable potential for HPV vaccination. However, the real-world effectiveness of HPV vaccination against anal high-grade lesions and cancer is yet to be shown. We investigated HPV vaccine effectiveness against anal high-grade squamous intraepithelial lesion (HSIL) or worse in a nationwide cohort including all Danish women aged 17-32 years during October 2006 to December 2021 (n = 968 881). HPV vaccinations and first occurrence of anal HSIL or worse were retrieved from nationwide registries. Women were considered vaccinated after first dose and classified by age at vaccination. Using Cox regression, hazard ratios (HRs) and 95% confidence intervals (CIs) were calculated for anal HSIL or worse according to vaccination status. During follow-up, the number of incident histological anal HSIL or worse cases was 37 in unvaccinated women, and less than 5 and 26 in women vaccinated at ages younger than 17 years and 17-32 years, respectively. The overall number of cancers was less than 5. Compared with unvaccinated women, the risk of histological anal HSIL or worse was reduced for women vaccinated at age younger than 17 years (HR = 0.30, 95% CI = 0.10 to 0.87). For women vaccinated at age 17-32 years, the hazard rate of anal HSIL or worse was 1.21 (95% CI = 0.73 to 2.03). This is the first study to demonstrate that HPV vaccination at a younger age is associated with substantially reduced risk of anal HSIL or worse in the general population.

Association between Human Papilloma Virus (HPV) genotype and protein of retinoblastoma expression in cervical cancer

Link of Video Abstract: https://youtu.be/5Q9QmV4Qmyg Background: Regarding prevalence among females worldwide, cervical cancer ranks fourth and has a high fatality rate. Cervical cancer growth is correlated with high-risk human papillomavirus (HPV) infection. Retinoblastoma protein (pRb) is a protein that interacts with E6/E7 oncoproteins to regulate cell proliferation, differentiation, and death. Mutations on the E6/E7 protein oncogenes of high-risk HPV can disrupt this relationship. This investigation aims to establish the association between pRb expression and the HPV genotype. Methods: This observational cross-sectional study examined 43 patients diagnosed with cervical cancer at Wahidin Sudirohusodo Hospital and its network. Samples were taken using consecutive random sampling based on established criteria. HPV genotype was examined using the polymerase chain reaction (PCR) method, while pRB expression was assessed by immunohistochemical examination. The relationship between the two variables was assessed using the Chi-Square test. Results: There were 5 (11.6%) patients with HPV type 16, 14 (32.6%) with HPV type 18, 12 (27.9%) with other types of HPV and 12 (27.9%) were negative for HPV. Examination of pRb expression showed that 33 (76.7%) samples had +2 expression, 7 (16.3%) had +1 expression, and 3 (7.0%) samples had no pRb expression. The pRb expression and HPV type were not significantly correlated in this study (p = 0.410). However, pRb expression and cervical cancer clinical stage were shown to be significantly correlated (p = 0.007). Conclusion: pRb expression is more prevalent when a malignancy is more advanced. In this investigation, no association between HPV genotype and pRb expression was discovered, despite oncogene mutations being involved in the regulation mechanism of cell proliferation, differentiation, and apoptosis.

Association between Human Papilloma Virus (HPV) genotype and mutant protein 53 (p53) expression in cervical cancer

Link of Video Abstract:https://youtu.be/fff-Unnud2w Background: Human Papilloma Virus (HPV) infection is the main cause of cervical cancer, a primary malignant tumor developing from squamous epithelial cells. HPV expresses oncoproteins E6 and E7, known to inactivate tumor suppressor proteins, one of which is protein 53 (p53). A promising biomarker for diagnosing and prognosis malignancies brought on by the HPV genotype is the identification of p53. This study investigates the association between p53 expression and HPV genotype in cervical cancer patients. Method: This observational cross-sectional study involved patients diagnosed with cervical cancer on histopathological examination who met the requirements. The polymerase chain reaction (PCR) method was used to determine the HPV genotype, and an immunohistochemistry analysis was used to assess the level of p53 expression. A Chi-Square test is utilized to evaluate the association between the two variables. Results: Of the 49 patients, there were 7 (14.3%) patients with HPV type 16, 13 (26.5%) with HPV type 18, 14 (26.5%) other types of HPV and 15 (30.6%) negatives for HPV. Examination of p53 expression showed that 17 (34.6%) samples had &lt;10% expression, 17 (34.6%) had 10-50% expression, and 15 (30.8%) samples had &gt;50% expression. There was no correlation between p53 expression and HPV genotype (p = 0.071). However, an association between p53 expression and cervical cancer's clinical stage was identified (p = 0.028). Conclusion: Increased cervical cancer stage can be associated with increased p53 expression. Thus, p53 can be used as a predictor of cervical cancer stage. However, in cervical cancer, p53 expression cannot be associated with the HPV genotype.

Epidemiology of oral human papillomavirus infection among diverse Chinese adults in typical areas of China: Findings from the DLCC study.

Human papillomavirus (HPV) infection is a predominant cause of oropharyngeal squamous cell carcinoma. However, there is limited knowledge about the epidemiology of oral HPV infections among adults in China. We collected data from a prospective cohort that enrolled participants in mainland China. A total of 9867 participants aged at least 20 years provided oral swab specimens in typical areas of China (Hebei and Guangdong provinces) in 2021. HPV DNA in oral exfoliated cells was tested using Nested Polymerase Chain Reaction and sequencing. Prevalence among sub-populations was compared. Multivariable logistic regression models were employed to assess possible factors influencing oral HPV infection. The overall prevalence of oral HPV infection was 3.0% [95% confidence interval (CI):2.68%-3.36%]. Among those infected, 1.3% (1.08%-1.53%) were infected with high-risk HPV types. Men had a higher age-standardized HPV infection prevalence (3.6%, 2.96-4.29%) compared to their female counterparts (2.7%, 2.35-3.12%). People in Hebei had a higher age- and sex-standardized prevalence (4.1%, 3.50%-4.70%) than those in Guangdong (2.2%, 1.80%-2.56%). Generally, men (odds ratio [OR] and 95% CI: 1.42, 1.09-1.85) and people in Hebei (2.01, 1.53-2.65) had higher odds of any type of HPV infection. Additionally, people living in urban areas had a 2.15-fold (1.43-3.26) higher odds of high-risk HPV infection. This study reveals a low prevalence of oral HPV infection with significant geographic and sex differences among Chinese population. This is the first study to report the epidemiological characteristics of oral HPV infection among Chinese adults in diverse geographic areas with large sample size.

Prevalence of high-grade HPV types among women in Astana, Kazakhstan (2018-2022)

Background: Cervical cancer is a significant public health concern worldwide, and human papillomavirus (HPV) is the leading cause of this disease. While there have been some studies on the prevalence of HPV in Kazakhstan, the country faces unique challenges in preventing and treating HPV-related diseases, including limited resources, a lack of awareness about HPV, and cultural attitudes toward sexual health. This research article aims to provide a comprehensive overview of HPV genotype distribution in Astana, Kazakhstan, for the last five years. Methods: A cross-sectional study was performed from 2018 to 2022. Study population: Kazakhstani women referring to “Olympus laboratories network” settings for HPV testing age 18-64. AmpliSens® Real-Time PCR kits for HPV genotyping 14 high-risk HPV types (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68) were performed. Results: This study revealed the increasing trend in HR-HPV prevalence among women for five sequential year periods. As a result, approximately 43% of women referred to the HPV testing were HR-HPV positive, with the most prevalent type being HPV 16 (20%), followed by HPV types 31, 52, 51. Conclusion: By examining the latest research on HPV and cervical cancer in Kazakhstan, we hope to contribute to developing effective public health policies and interventions to help reduce these diseases' burden in this country. Furthermore, the database will be established for future large-scale studies after vaccination implementation.

First international proficiency study on human papillomavirus testing in cervical cancer screening

BackgroundAlthough cervical screening using Human Papillomavirus (HPV) testing is globally recommended public health policy, there has been no international proficiency studies specifically targeting HPV testing for cervical screening. ObjectiveTo obtain the first global overview of the current proficiency of HPV testing services for cervical cancer screening. Study designA coded proficiency panel of 12 samples containing HPV types 16, 18, 31, 33, 45, 52, 58 or 35/39/51/56/59/68 in human DNA in varying amounts as well as control. Datasets detecting at least a) 10 International Units (IU) of HPV16 and 18, b) 1000 IU of HPV types 31, 33, 45, 52, 58 and c) having no false positives were considered proficient. ResultsIn total, 84 laboratories worldwide submitted 158 datasets (some laboratories used >1 HPV testing platform). Of those, 122 (77%) were 100% proficient. Only 14/158 datasets (9%) contained false positive results. Comparison of results with assays approved by the Food and Drug Administration (FDA) suggest that future proficiency requirements should also accommodate assays detecting only 100 IU of HPV16/18. A pool of low oncogenicity HPV types that contributed very little to sensitivity, but adversely affected specificity, was detectable by most datasets. ConclusionInternationally recognized proficiency studies of HPV screening, traceable to international standards, provided an overview of current testing performance. There was a high level of proficiency in terms of sensitivity and few false positives, but specificity was not optimal and further research on optimal specificity of HPV screening tests may be warranted.

Low presence of papillomavirus and its lack of correlation with clinicopathological factors in breast cancer: a case control study.

Breast cancer is currently the most commonly diagnosed neoplasm in women worldwide. There is evidence that human papillomavirus (HPV) infection may play a key role in breast cancer aggressiveness, but results are conflicting across studies. The aim of this study was to investigate the presence of the HPV viral genome in benign and malignant breast tissue samples and its clinicopathological characteristics of cancer. In this case-control study, 100 formalin-fixed paraffin-embedded (FFPE) of breast cancer and 100 blocks of non-cancerous breast tissue were selected as a control group from the pathology department of Imam Khomeini Hospital in Ahvaz from 2020-2022. The presence of HPV was detected using nested PCR including MY09/11 primers and sequencing were performed for virus genotyping. The present study enrolled 100 subjects each in two cancer and control groups with a mean age of 52.81±13.23 and 35.77±11.65, respectively. The risk of cancer in HPV-infected patients is almost 5 times higher than in HPV-negative individuals, it is not statistically significant (OR =4.99, 95% CI 0.35 to 72.15, p=0.238). The prevalence of HPV in the cancer and control groups was 7% and 1%, respectively and HPVs detected in two groups were of the HPV 16 genotype. Although the chance of ER and PR expression, lymphvascular involvement, perineural invasion, and higher tumor grade was higher in HPV-positive subjects than in HPV-negative subjects, this was not statistically significant (OR>1, p>0.05). Based on studies reporting the existence of sequences of different high-risk HPV types (oncogenes) in breast cancer tissues, this study confirmed the hypothesis of a possible infectious cause in the development of breast cancer. So far, however, the results have been controversial and inconclusive. Further studies with large sample sizes are needed to demonstrate the link between HPV and breast cancer.

Assessment of Human Papillomavirus Non-16/18, Type-Specific Risk for Cervical Intraepithelial Neoplasia Grade 3 or Worse Among Women With Cervical Atypical Glandular Cells.

To evaluate the risk for cervical intraepithelial neoplasia grade 3 (CIN 3) or worse (including adenocarcinoma in situ [AIS] and invasive cervical cancer) associated with non-16/18 human papillomavirus (HPV) types (other HPV) among women with atypical glandular cells (AGC) in cervical cytology. This population-based cohort study evaluates the risk of CIN 3 or worse associated with other HPV types. Human papillomavirus genotyping was performed on Pap tests collected in Sweden from 341 women with AGC that were positive for other HPV types from February 17, 2014, to December 31, 2018. The women were followed for histopathologic outcomes using comprehensive registry linkages until December 31, 2019. Cumulative incidence proportions of CIN 3 or worse by specific HPV type were calculated using 1-minus Kaplan-Meier function. Hazard ratios (HRs) for CIN 3 or worse were generated using multivariate Cox regression. Of 341 women, 134 (39.3%) had CIN 3-AIS, but there were only five (1.5%) women in the cohort with invasive cervical cancer. Human papillomavirus 45 preceded 80.0% of invasive cervical cancer cases. Among women positive for HPV33, 82.9% (95% CI 58.0-97.3%) had CIN 3 or worse during follow-up. Positivity for HPV31 conferred the highest HR for CIN 3 or worse relative to other types, both in primary cytology and primary HPV screening (HR 2.71, 95% CI 1.47-5.00 and HR 3.41, 95% CI 1.95-5.96, respectively). Among non-16/18 HPV types in AGC, HPV31 and 33 had the highest risk for CIN 3 or worse, whereas most of the women with invasive cancer were positive for HPV45. Extended HPV genotyping may be helpful for the management of AGC.

A human papillomavirus whole genome plasmid repository: A resource for HPV DNA quality control reagents.

Well characterized reference reagents are useful for assay validation, proficiency/competency assessment, daily run controls, and to improve inter-laboratory comparisons. Synthetic human papillomavirus (HPV) DNA fragments and plasmid clones are available, but synthetic fragments include limited segments of the HPV genome and many HPV plasmids have interrupted coding regions or contain partial genomes. As a result, they are not compatible with all HPV DNA detection and typing assays. To address this need, we are establishing an HPV plasmid repository of HPV clones containing the whole genome of each type with no interruptions in coding regions. To date, HPV plasmid clones for 16 HPV types, (including all vaccine types and 14 types in clinical assays: HPV6, 11, 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66 and 68) have been constructed using a Gibson assembly method and validated by sequencing and the Novaplex HPV typing assay. The newly constructed HPV whole genome plasmids can serve as a quality control reagent resource for HPV DNA assays and are available for public health and research laboratories.

Effect of Pap Smear Cytology, HPV Genotyping on the Concordance of Colposcopy and Conization Results.

To evaluate the conization results performed due to human papillomavirus (HPV), smear, colposcopy results or clinician's decision and determine the factors that predict ≥CIN2. Retrospective comparative study. Place and Duration of the Study: Department of Gynaecology and Obstetrics, Dr. Abdurrahman Yurtaslan Ankara Oncology Training and Research Hospital, Ankara, Turkey, between January 2011 and December 2021. Women with known HPV results who underwent conization in the Gynaecology clinic were retrospectively included. Age, HPV genotypes, conization, and colposcopy results of the patients were recorded. Patients were divided into two groups as those with and without ≥CIN2 and compared in terms of clinicopathological features. Four hundred and twenty eight (82.8%) of the 517 patients were premenopausal and perimenopausal, and 89 (17.2%) of the patients with a median age of 42 years (range: 30-65 years) were postmenopausal. While 374 were HPV 16/18 positive, 143 were non-16/18 HPV positive. Conization result was normal in 202 (39.1%) patients, CIN1 in 129 (25.0%) patients, and CIN 2-3 in 186 (36.0%) patients. In the HPV 16/18 positive group, conization result was normal in 38.2% of patients, CIN1 in 20.9%, and CIN 2-3 in 40.9%; these rates were 41.3%, 35.7%, and 23.1% in the HPV-other group, respectively (p <0.001). In the logistic regression model, age, HPV type (16/18), and smear cytology results (≥ASC-US) were tested as independent factors predicting ≥CIN2. HPV 16/18 positivity and smear cytology result (≥ASC-US) were the factors predicting ≥CIN2. Smear and HPV genotyping can make an important contribution to detecting false <NIC2 results as a result of colposcopy. CIN, Colposcopy, Conization, Cervix, Cervical cancer, Neoplasia, HPV.