High-risk Human Papillomavirus Genotypes Research Articles

Cytological DNA methylation for cervical cancer screening: a validation set.

In a previous training set with a case-controlled design, cutoff values for host EPB41L3 and JAM3 gene methylation were obtained for the detection of cervical intraepithelial neoplasia (CIN) 2 or more severe lesions (CIN2+). This validation trial was conducted to evaluate the role of DNA methylation in screening for CIN2+ by cervical cytology among unselected participants. From June 1, 2019, to September 1, 2019, in our study center, we collected liquid-based samples from cervical swabs for methylation assays and hrHPV testing in eligible patients. The primary endpoint was the diagnostic accuracy of DNA methylation and hrHPV genotyping for CIN2+ according to confirmed histology results. Among 307 participants, compared with hrHPV testing, the methylation assay for CIN2+ had lower sensitivity (68.7% versus 86.1%, p=0.002) but higher specificity (96.7% versus 0.696, p<0.001). The methylation assay also had favorable sensitivity and specificity in patients with negative hrHPV testing (56.3% and 96.9%) and in patients with cervical adenocarcinoma (73.7% and 92.7%). DNA methylation had higher specificity than the hrHPV assay (100.0% versus 44.4%, p<0.001) for identifying residual CIN2+ in patients without residual lesions. Positive cervical DNA methylation was associated with a diagnostic probability of endometrial carcinoma (odds ratio 15.5 [95% confidence interval 4.1-58.6]) but not of ovarian epithelial carcinoma (1.4 [0.3-6.5]). The host EPB41L3 and JAM3 gene methylation assay in cervical cytology had favorable diagnostic accuracy for CIN2+ and was highly specific for residual CIN2+ lesions The methylation assay is a promising triage tool in hrHPV+ women, or even an independent tool for cervical cancer screening. The methylation status in cervical cytology could also serve as a prognostic biomarker. Its role in detecting endometrial carcinomas is worthy of further exploration.

Variability of High-Risk Human Papillomavirus and Associated Factors among Women in Sub-Saharan Africa: A Systematic Review and Meta-Analysis.

Sub-Saharan Africa (SSA) carries the highest burden of high-risk human papillomavirus (HR-HPV) in the world, driven by, and together with, HIV infection. This systematic review aimed to identify HR-HPV genotypes and their associated factors among women in SSA. A systematic review and meta-analysis of studies conducted in SSA on HR-HPV was conducted. Standard electronic databases were searched. R software version 3.6.0 was used for meta-analysis, with p < 0.05 considered statistically significant. We included 28 articles with a total of 22,652 participants. The overall pooled prevalence of HR-HPV genotypes was 55.13%, albeit high heterogeneity between studies. The overall pooled prevalence of HR-HPV genotypes in HIV-positive individuals was 75.51%, compared to 52.97% in HIV-negatives (OR = 4.68 (0.71-30.76)). HPV 16 (18%), 35 (10.12%), 52 (9.98%), 18 (9.7%) and 45 (6.82%) genotypes were the most prevalent. Twelve studies identified the most frequently reported risk factors associated with HR-HPV, with HIV infection (66.66%), multiple sexual partners (41.66%) and young age (41.66%) being the most reported risk factors. The combined prevalence of HR-HPV genotypes among women in general and HIV-infected women in particular remains high in SSA. The presence of several genotypes not covered by the vaccine is remarkable and suggests the need for revision of current vaccination policies to prevent HR-HPV infections.

Detection of cervical high-grade squamous intraepithelial lesions and assessing diagnostic performance of colposcopy among women with oncogenic HPV

BackgroundHPV screening tests may improve cervical cancer risk stratification and better guide decisions about follow-up with colposcopy/biopsy. This study aimed to estimate the risk of cervical intraepithelial neoplasia grade 2 or worse (CIN2+) among women with oncogenic HPV types and evaluate the performance of colposcopy in the diagnosis of histologic CIN2 + at Putuo Hospital, Shanghai, China.MethodsThis cross-sectional survey was conducted from February 2020 to December 2022 among women who were referred to colposcopy. Women with high-risk (HR) HPV-positive, cytology testing and colposcopy-directed biopsy were included.ResultsUnivariate and multivariate analysis indicated that high-grade colposcopic impression ((OR, 17.61%, 95%CI: 11.54–26.85%) was associated with the highest risk for detecting CIN2+, followed by HSIL + cytology (OR, 6.90%, 95%CI: 3.56–13.37%) and HPV16/18 positive (OR, 2.91%, 95%CI: 2.12–3.99%). Overall, CIN2 + was detected in 14.6% of 2007 women. HPV16/18 had higher CIN2 + risks than other HR-HPV genotypes (30.1% vs. 10.2%, P<0.001). Among women with low-grade cytology, 24.1% had CIN2+, and the risks for HPV16/18 (58.2%) were higher than for other HR-HPV(16.8%). For those with high-grade cytology, there was no significant difference between HPV groups ( 75.0% vs. 72.9%, P > 0.05). The diagnostic performance of colposcopy in diagnosis of CIN2 + by senior and junior colposcopists was comparable.ConclusionsThe results indicated that referral to colposcopy is recommended in managing women with HR-HPV positive, and colposcopic impressions provide key clues for identification CIN2+.

HPV vaccination in Kenya: a study protocol to assess stakeholders' perspectives on implementation drivers of HPV vaccination and the acceptability of the reduced dose strategy among providers.

Cervical cancer is the leading cause of cancer-related deaths among Kenyan women. Persistent infection with high-risk oncogenic Human papillomavirus (HPV) genotypes is a necessary cause of cervical cancer. HPV vaccines are safe, durable, and efficacious in preventing incident HPV infections. In Kenya, despite efforts to increase HPV vaccination, coverage remains low. We sought to assess: (1) barriers and facilitators of HPV vaccination from the perspective of adolescent girls and young women (AGYW), their guardians as well as stakeholders involved in HPV vaccine delivery, and (2) the acceptability of the single dose of the HPV vaccination among healthcare providers (HCPs). Our study is nested within the KENya Single-dose HPV-vaccine Efficacy study (KEN SHE) that sought to test the efficacy of single-dose bivalent (HPV 16/18) and single-dose nonavalent (HPV 16/18/31/33/45/52/58/6/11) vaccination. We are conducting this study in Kiambu, Nairobi, and Kisumu counties. In these counties, we are interviewing stakeholders (n = ∼25), selected based on their role in HPV vaccination at the county and national levels. Interviews are audio recorded and conducted in English or Swahili. The semi-structured interview guides were designed based on: (1) the Theoretical Domains Framework (TDF) for AGYW and guardians and (2) the Consolidated Framework for Implementation Research (CFIR) for other stakeholders. The Theoretical Framework of Acceptability (TFA) was leveraged to design the survey administered to HCPs (n = ∼309) involved in HPV vaccination. We will develop a codebook based on emerging codes from the transcripts and constructs from the TDF and CFIR. Emerging themes will be summarized highlighting similarities and differences between and within the different stakeholder groups and counties. Descriptive statistics and a χ2 test will be used to assess the distribution of responses between the different sites and regression analysis will be used to assess factors associated with high acceptability of the single-dose strategy while controlling for confounding variables. Our study will describe key barriers and facilitators that affect HPV vaccination from the perspective of multiple stakeholders as well as insights on the perspective of HCPs towards the single-dose strategy to inform the designing of strategies to increase HPV vaccination uptake in Kenya and comparable settings.

Does infection of multiple HPV high-risk genotypes carry a higher malignant potential than a single genotype in uterine cervix? A retrospective study.

86 Background: The high-risk genotype of human papilloma virus (HPV) has been known as the most important carcinogen in uterine cervical carcinoma. It remains unclear that the infection with multiple high-risk genotypes carry a higher malignant potential risk compared with a single genotype in uterine cervix. Methods: This is a retrospective study of 695 patients underwent conization at Kyungpook National University Hospital in South Korea from 2012 to 2023. Participants were required to have undergone an HPV test by cervical swab within 12 months prior to the surgery. Patients with a negative result or a previous history of cancer were excluded. Additionally, patients older than 50 years were excluded to reduce bias. We included patients with the most common genotype in the cohort with only one high-risk genotype, and patients with multiple high-risk genotypes including the most common genotype. We categorized them based on pathologic results. Patients with HSIL, carcinoma in situ (CIS), invasive carcinoma were classified as ≥HSIL, while those with CIS, invasive carcinoma were classified as ≥CIS. The HPV test was performed using DNA microarray or real-time polymerase chain reaction, can detect 18 high-risk genotypes and 14 low-risk genotypes. The infection status of low-risk HPV and the vaccination for HPV were not considered in this study. The statistical analysis was conducted using SPSS (version 26). Results: A total of 146 patients younger than 50 years found to be infected with HPV high-risk genotype 16 or 16 with other high-risk genotypes. The HPV 16 single infection and multiple infection were found in 79 (54.1%), 67 (45.9%), respectively. Among patients with HPV 16 single infection, 61 (77.2%) were diagnosed with ≥HSIL and 45 (57.0%) were ≥CIS. In the HPV 16 multiple infection group, 56 (83.6%) were diagnosed with ≥HSIL and 34 (50.7%) were ≥CIS. There was no statistically significant correlation based on the HPV 16 single or multiple infection status for either ≥HSIL (HPV 16 single infection vs. multiple infection, odds ratio (OR)=0.666 [95% CI=0.289-1.531], p=0.407) or ≥CIS (HPV 16 single infection vs. multiple infection, OR=1.285 [95% CI=0.668-2.471], p=0.507). Conclusions: Among young women with only a single HPV high-risk genotype, the genotype 16 was the most common. Compared to the infection group with a single HPV high-risk 16 genotype, the malignant potential was not found to be significantly increased in the infection group with multiple HPV high-risk genotypes, including HPV genotype 16. [Table: see text]

Prevalence of Human Papillomavirus (HPV) Genotypes among Women During 2015-2020 in Mashhad, Iran.

Cervical cancer is the fourth most common cancer in women, and human papillomavirus (HPV) is the leading cause of cervical cancer. Cervical cancer screening and HPV vaccination are important in the incidence of cervical cancer. This study was performed on Liquid Base Cytology (LBC) samples of 1214 women in Mashhad who were referred for cervical cancer screening in 2015-2020. Samples were examined by Single-Step PCR and Reverse Line Blot for HPV genotyping. 386 women (31.8%) were HPV PCR positive. HPV genotyping of 277 samples showed that HPV 31 (3%), 16 (2.5%), 51 (2.2%), 18 (2%), and 66 (1.8%) were the most prevalent high-risk HPV (hrHPV) genotypes. Among low-risk HPV (lrHPV) genotypes, HPV 6 (9.2%), 53 (4.7%), and 42 (2.8%) were the most common genotypes. The range of multiple infections varied between two to eight genotypes and the prevalence of multiple HPV infections (12.4%) was higher than single infections (10.4%). For women with single HPV infections, HPV 31 and 66 were equally the most common hrHPV genotypes, followed by HPV 16 and 39. In women with multiple HPV infections, HPV 31 was the most common hrHPV genotype, followed by HPV 51 and 16. For both the single and multiple HPV infections, HPV 6 was the most common lrHPV genotype, followed by HPV 53 and 42. In conclusion, due to the high prevalence of HPV single and multiple infections, the need for governmentally supported HPV vaccination and through cervical cancer screening should be emphasized to prevent cervical cancer.

Prevalence of human papillomavirus infection among women at a gynaecological clinic in Tlemcen, Algeria.

High-risk human papillomavirus (HR-HPV) is recognized as the etiologic agent responsible for cervical cancer, ranking as the second most prevalent cancer among women in Algeria with an incidence rate of 10.4 per 100,000. The primary aim of this study was to conduct a preliminary prospective investigation into the detection of HR-HPV infections in Tlemcen, Algeria, where screening is exclusively based on cytology. A total of 130 cervical swabs were analysed in this study. HPV detection was performed utilizing the Cobas® 4800 test, incorporating polymerase chain reaction (PCR) for individual genotyping of HPV-16 and HPV-18, as well as pooled detection of 12 other commonly occurring HPVs. The findings revealed that out of the 130 samples, 28 tested positive for HR-HPV, resulting in a prevalence rate of 21.5%. Among these cases, five infections demonstrated the coexistence of HPV16 with other HR-HPV genotypes. The prevalence of HPV16 infections was determined to be 28.6% (8/28), whereas 68% of infections (19/28) were attributed to other HR-HPV genotypes. These observations indicate that HPV16 was not the prevailing genotype. Consequently, these results underscore the necessity for a larger-scale study with an expanded sample size encompassing cytology and HPV testing. Such an investigation would be invaluable in facilitating the development of a national prevention program to effectively control cervical cancer.

Prevalence and human papillomavirus (HPV) genotype distribution in Suzhou, China

ABSTRACT Persistent infection with high-risk human papillomavirus (HR-HPV) is a known pathogenic factor of cervical cancer. To develop scientific guidance for cervical cancer screening and HPV vaccination, we analyzed HPV genotypes in Suzhou City, China. This study utilized data from the cervical cancer screening project in Suzhou from 2016 to 2021. A total of 444,471 female residents who voluntarily underwent HPV testing were included in the study. The overall HR-HPV prevalence was 10.2%. The three most common HR-HPV genotypes were HPV52 (2.81%), HPV58 (1.64%), and HPV16 (1.46%). The rate of HPV infection increased with age. Having a junior school education or higher was a protective factor compared to having an education level below junior school. The overall HPV infection rate showed a downwards trend from 2016 to 2021. HPV16 exhibited the fastest annual decline rate, followed by HPV18. As the severity of cervical intraepithelial neoplasia increases, the detection rate of HPV infection significantly increased. In conclusion, in addition to cervical cancer screening, it is important to pay attention to health promotion and education for low-educated women aged 45–59. Considering the distribution of HPV genotypes, prioritizing the administration of high-valency HPV vaccines to local seventh-grade female students is recommended.

High-risk human papillomavirus genotyping in women with atypical squamous cells of undetermined significance

We conducted a prospective study to evaluate the prevalence of high-risk human papillomavirus (hr-HPV) positivity in women with atypical squamous cells of undetermined significance (ASC-US). Additionally, we assessed the association of hr-HPV positivity with the pathology of high-grade squamous intraepithelial lesions or worse (HSIL+) and the risk of subsequent detection of squamous intraepithelial lesions. A total of 376 women were included, with 242 (64.4%) exhibiting hr-HPV positivity. The predominant HPV genotypes were 16, 52 and 58. Factors associated with the immediate detection of HSIL+ pathology included a colposcopic impression of high-grade lesions, hr-HPV positivity, HPV 16 positivity, HPV 18 positivity, HPV 58 positivity, age less than 40 years, and biopsy of two or more pieces. However, only the first three factors were statistically significant in multivariate analysis. Among the 291 women who continued surveillance for 6 months or more, the median follow-up period was 41.8 months (interquartile range [IQR] 26.5–54.0). The prevalence of subsequent HSIL in women with hr-HPV positivity versus negativity was 3.6% versus 0.98%, respectively. The median time to the subsequent detection of SIL was 28.7 months (IQR 14.9–41.7). In conclusion, women with ASC-US in our study had a high proportion of hr-HPV positivity. Type-specific HPV testing could play a pivotal role in the development of specific management protocols for women with ASC-US.Clinical trial registration: https://thaiclinicaltrials.org, TCTR20161017002.

Evaluation of the Roche MagNA Pure 96 nucleic acid extraction platform for the SPF10 line probe assay system.

To evaluate the MagNA Pure 96 (MP96) nucleic acid extraction system as an alternative for cervical specimen processing for human papillomavirus (HPV) genotyping detection by reverse hybridization line probe assay (LiPA-25), compared to the well-established extraction system MagNA Pure LC 2.0 (MPLC). A total of 200 cervical samples preserved in ThinPrepCyt® solution were extracted by MP96 and MPLC, respectively, and then purified nucleic acids were amplified using the SPF10 PCR primer set. Amplification products were subjected to SPF10-DNA enzyme immunoassay (DEIA) and LiPA-25. The concordance between different extraction methods in this study was reflected in the comparison of the results of the DEIA and LiPA-25. Agreement of HPV-positive results (DEIA) was 97.5% (Kappa=0.932). Pair-wise analyses of either HPV grouping (any HPV genotypes, any high-risk HPV genotypes, any low-risk HPV genotypes, any nonavalent vaccine-targeted HPV, and any non-vaccine-targeted oncogenic HPV genotypes) identified >95% agreement (all Kappa>0.900). For the two extraction methods, there was no statistical difference (chi-square test: P=0.690) for single versus multiple genotypes, and concordant, compatible, and discordant genotypes were observed in 87.0%, 9.5%, and 3.5% of 200 samples, respectively. HPV genotyping results of the MPLC system and the MP96 system showed a high degree of concordance. Combined with the advantages of high-throughput and anti-contamination of MP96, the MP96 extraction system could be more suitable for the testing of samples in future studies.

Testing for Human Papillomaviruses in Urine, Blood, and Oral Specimens: an Update for the Laboratory.

Twelve high-risk alpha human papillomavirus (HPV) genotypes cause approximately 690,000 cancer cases annually, with cervical and oropharyngeal cancer being the two most prominent types. HPV testing is performed in laboratory settings for various applications of a clinical, epidemiological, and research nature using a range of clinical specimens collected by clinicians or by individuals (self-collected specimens). Here, we reflect on the importance and justification of using the right test for the right application and provide practical updates for laboratories either participating in or anticipating involvement in HPV testing in three specimen types, namely, urine, blood, and oral specimens, which are considered "alternative" specimens by many. In addition to clinician-collected cervical samples and self-collected cervicovaginal samples, first-void urine is emerging as a credible specimen for HPV-based cervical cancer screening, triage of HPV screen-positive women, monitoring HPV vaccine impact, and HPV testing in groups for which a less invasive sample is preferred. Detection of cell-free DNA (including HPV DNA) in blood has great promise for the early detection of HPV-attributable oropharyngeal cancer (HPV-AOC) and potentially other HPV-driven cancers and as an adjunct prognostic marker in long-term tumor surveillance, including treatment response. The moderate sensitivity of HPV testing in oral rinses or swabs at HPV-AOC diagnosis prevents its use in HPV-AOC secondary prevention but represents a promising prognostic tool in HPV-AOC tertiary prevention, where the HPV persistence in oral rinses throughout treatment may predict early HPV-AOC recurrences and/or the development of secondary HPV-AOC. The increasing sophistication of specific collection devices designed for alternative samples and the enhanced precision of novel molecular technologies are likely to support the evolution of this field and catalyze potential translation into routine practice.

Design and Analytical Evaluation of a Rapid Plasma Screening Assay for Circulating Human Papillomavirus DNA via Thermostable Enzyme Chemistries.

Infection with oncogenic strains of human papillomavirus (HPV), such as HPV-16 and HPV-18, can lead to malignant progression and tumorigenesis. As an adjunct to traditional invasive tissue sampling methods, the use of modern thermostable enzyme chemistries can aid in the development of innovative assay workflows to extract and detect circulating HPV DNA (cHPV-DNA) in liquid biopsies. In this work, we first successfully generated a model system to replicate fragmented cHPV-DNA in human plasma. Using this model system, we designed a novel thermostable enzyme chemistry-based cHPV-DNA assay for rapid clinical HPV screening and robustly evaluated its analytical assay performance. Our findings demonstrated that the use of thermostable enzymes provided faster cHPV-DNA extraction and amplification, leading to an overall three-fold improvement in overall assay time as compared to the current standard assay workflow and achieving clinically relevant levels of analytical specificity, sensitivity, and precision for accurate cHPV-DNA detection with excellent 100% sensitivity and specificity in contrived human plasma specimens. In summary, we have devised a rapid laboratory workflow to facilitate the emerging use of liquid biopsies for minimally invasive, rapid, and scalable HPV DNA testing. With facile assay modifications, our thermostable enzyme-based cHPV-DNA assay can be utilized for the detection of other clinically high-risk HPV genotypes.

Polymorphism of KIR Genes in Women with Human Papillomavirus Infection

Molecular biology screening techniques for early detection of human papillomavirus (HPV) infection in the National CervicalUterine Cancer Program in Cuba provide the opportunity to treat premalignant lesions and prevent progression to cervicaluterine cancer. Objectives: To identify 14 high-risk HPV genotypes in women aged 30 to 50 with negative previous cytology and to identify the polymorphism of killer immunoglobulin-like receptor (KIR) genes in a subsample of HPV-positive women. Methods: HPV screening tests were performed on 3,115 women using the COBAS 4800 system with the HPV COBAS kit (Roche, Germany). For KIR gene typing, 60 randomly selected HPV-positive women were analyzed using a molecular method based on hybridization probes on a LUMINEX flow analyzer with the LIFECODES KIR-SSO typing kit (Immucor, USA). Results: 295 (9.5%) women tested positive for one of the 14 high-risk genotypes. The highest percentage of positive women was found among those aged 30 to 39 years (12.0%). Fourteen women had coinfection with HPV16 or HPV18 along with another highrisk genotype. There was a high frequency of genes encoding activating receptors such as KIR 2DL1 (98.3%), KIR 2DL3 (98.3%), and KIR 2DS4*all full length (93.3%). Conclusions: The prevalence of HPV in Cuban women with normal cytology is lower than the global average, with activating KIR genes predominating among positive cases.

High-risk human papillomavirus infection among Nigerian women: A systematic review and meta-analysis.

We conducted a systematic review and meta-analysis to determine the prevalence of high-risk human papillomavirus (hrHPV) infection and its associated risk factors among Nigerian women. Databases including PubMed, Web of Science, Scopus, and CINAHL were searched for studies published between 01 January 2001 and 31 December 2022, that had reported hrHPV infection and associated risk factors among women in Nigeria from ages of 25 to 65 years. Of the 136 records initially retrieved, 18 were eligible for analysis. The prevalence of hrHPV genotypes was 25%, and for hrHPV 16 and 18, were 9% and 10%, respectively. The prevalence of hrHPV among HIV+ve women was 71%. The most common risk factors for hrHPV were age at coitarche and multiple sex partners. hrHPV prevalence is high in women in Nigeria and common among those HIV+ve. Rapid screening for hrHPV genotypes is recommended, and multivalent HPV vaccines should be considered for women.

First study of genital HPV infection among women living with HIV recruited from May to September 2018 in Eastern Algeria.

Genital HPV infection is recognized worldwide as one of the most common, if not the most common, sexually transmitted infection. Several studies have shown a higher prevalence of HPV infection in women living with HIV (WLWH). We aimed to determine the prevalence of HPV, the circulating genotypes, and its association with risk factors among Algerian WLHIV. Cervical specimens were collected from 100 WLHIV. The detection of HPV infection was performed by Roche Linear Array test. The overall prevalence of HPV infection was determined at 32%, all including high risk HPV (HR-HPV). We found a great diversity of HR-HPV genotypes, the most common individually were HPV52, HPV16, HPV18 and HPV58, The highest prevalence was found in groups of genotype 52, 25%. HPV16 and 18 were less common (16%). Cervical cytology was abnormal in 66% (81.3% in the HPV + population), with a predominance of inflammatory lesions (75% HPV +), The most important determining risk factor for HPV infection in this series was a low CD4 T cell count <200/mm3 found in 72% of the HPV positive participants. Our study is an initial database, to be completed by a multicenter study to determine the most frequent genotypes in Algerian WLHIV, in order to discuss the introduction of the vaccine against HPV in Algeria, especially among WLHIV.

Evaluating the performance of anal cytology and high-risk HPV genotyping for detecting anal HSIL in a clinic-based sample of people living with and without HIV in Puerto Rico.

Given the disproportionately elevated anal cancer risk in high-risk populations, it is important to assess the performance of commonly used anal cancer screening tools to improve the effectiveness of detection and treatment methods. This study evaluates 1) the concordance between anal cytology and histology results and 2) the performance of cytology and high-risk human papillomavirus (HR-HPV) genotyping as screening tools for detecting histologically confirmed anal high-grade squamous intraepithelial lesions (HSIL). Data from the Anal Neoplasia Clinic in Puerto Rico (2014-2021; n = 466) was used. The clinical performance of anal cytology and HR-HPV genotyping to detect HSIL was compared to the gold standard: high-resolution anoscopy-guided biopsy. Sensitivity, specificity, positive predictive value, negative predictive value, and κ coefficients were calculated. A total of 66.95% of the patients were men, 74.0% were people living with HIV, 76.2% had anal HR-HPV infection, and 40.34% had histologically confirmed anal HSIL. The weighted κ statistic between the tests (cytology and histology) was 0.25 (p < .001). The sensitivity and specificity of cytology alone to detect anal HSIL were 84.3% (95% confidence interval [CI], 78.3%-89.1%) and 36.0% (95% CI, 30.3%-42.0%), respectively. Anal HR-HPV genotyping had higher sensitivity (92.2%; 95% CI, 87.4%-95.6%) and similar specificity (34.8%; 95% CI, 29.2%-40.7%) compared to cytology. The two tests combined (positive results following cytology or HR-HPV test) improved sensitivity to detect anal HSIL (97.9%; 95% CI, 94.8%-99.4%), but specificity was compromised (19.2%; 95% CI, 14.7%-24.4%). Although HR-HPV genotyping improved the detection of anal HSIL, HR-HPV testing had lower specificity than anal cytology alone.

Detection of human papillomavirus in archival bladder and ovarian cancer samples

ProblemHuman papillomavirus (HPV) is a well-established causative agent of cervical cancer. However, its role in the development of bladder and ovarian cancer is under debate. Current study evaluated the role of high-risk HPV genotypes 16 and 18 in triggering bladder and ovarian carcinomas. MethodsA total of 80 formalin-fixed and paraffin-embedded (FFPE) archival tissue specimens of both bladder (n = 40) and ovarian carcinoma (n = 40) were tested for the presence of HPV integrants and HPV 16 genotype and HPV 18 genotype. Results and conclusionThe PCR results showed that 5% (2/40) of bladder cancer samples were HPV positive with equal prevalence of HPV 16 in both sexes (1:1) while 22.5% (9/40) of ovarian cancer samples came positive for HPV with only one positive for HPV 16. These results indicate association of HPV with bladder and ovarian carcinomas, HPV 16 genotype being more prevalent than 18. The study emphasizes that HPV is a serious issue in low- and middle-income countries where existing methods of prevention are still inadequate or costly. Timely HPV screening can help lower the burden of HPV-induced carcinomas. Furthermore, systematic HPV vaccination program among the young population is paramount.

Development of a simplified and cost-effective sample preparation method for genotyping of human papillomavirus by next-generation sequencing.

High-risk human papillomavirus (HPV) infection is the most common cause of cervical cancer, but low-risk HPV strains can sometimes also be involved. Although HPV genotyping techniques used in clinical diagnosis cannot detect low-risk HPV, next-generation sequencing (NGS) can detect both types. However, DNA library preparation is complicated and expensive. The aim of this study was to develop a simplified, cost-effective sample preparation procedure for HPV genotyping based on next-generation sequencing (NGS). After DNA extraction, a first round of PCR was performed using modified MY09/11 primers specific for the L1 region of the HPV genome, followed by a second round of PCR to add the indexes and adaptors. Then, the DNA libraries were purified and quantified, and high-throughput sequencing was performed using an Illumina MiSeq platform. The sequencing reads were compared with reference sequences for HPV genotyping. The limit of detection for HPV amplification was 100 copies/µl. Analysis of the correlation of pathological cytology with the HPV genotype in individual clinical samples showed that HPV66 was the most common genotype found in the normal stage, whereas HPV16 was the main genotype found in low-grade squamous intraepithelial lesions, high-grade squamous intraepithelial lesions, and cervical cancer. This NGS method can detect and identify several HPV genotypes with 92% accuracy and 100% reproducibility, and it shows potential as a simplified and cost-effective technique for large-scale HPV genotyping in clinical samples.

Immediate histopathologic correlation in Turkish population with negative cytology and high-risk human papillomavirus positivity: A retrospective analysis of high-risk human papillomavirus genotype and stratified by age.

According to the American Society of Colposcopy and Cervical Pathology (ASCCP) recommendations, regardless of age, women with high-risk infections other than human papillomavirus 16/18 positivity (other hrHPV) and negative cytology should not be referred directly to colposcopy. Several studies compared detection rates of ≥high-grade squamous intraepithelial lesion (HSIL) between HPV 16/18 ± 45, and other hrHPV types on colposcopic biopsy. We designed a retrospective study to determine the presence of ≥HSIL in colposcopic biopsy in women with negative cytology and hrHPV positivity during the years 2016-2022. HPV 16/18/45 had a PPV of 43.8%, while other hrHPV types had a PPV of 29.1% for a tissue diagnosis of ≥HSIL. For a tissue diagnosis of ≥HSIL detection, there was no statistically significant difference between the PPV of other hrHPV and HPV 16/18/45 in patients ≥30. There were only two cases with a tissue diagnosis of ≥HSIL in the other hrHPV group of women under 30 years of age. We suggested that the follow-up recommendations of ASCCP for patients above the age of 30 with negative cytology and other hrHPV positivity may not be fully applicable to countries like Turkey with a different healthcare environment. Referring to patients ≥30 who had other hrHPV positivity and negative cytology to direct colposcopy may be clinically beneficial, particularly in populations where a colposcopic examination is easy and inexpensive.

The role of multiple high-risk human papillomavirus infections for cervical biopsies and findings in colposcopic procedures

The clinical outcome of high-risk HPV (hr-HPV) infection varies according to genotype(s). Patients may harbor either one single hr-HPV (s-HPV) or multiple HPV (m-HPV) genotypes. Recently, the relationship between m-HPV infections and high-grade dysplasia has been investigated, and controversial results have been obtained. Therefore, the clinical significance of m-HPV is not clear. This study aimed to evaluate which group is associated with higher grade dysplasia by analyzing colposcopic punch biopsies. A total of 690 patients who were scheduled for a diagnostic excisional procedure between April 2016 and January 2019 due to the detection of high-grade cervical intraepithelial neoplasia (CIN 2/3) in colposcopy were included. Patients who were not scheduled for colposcopic examination or cervical punch biopsy, or who were scheduled for an excisional procedure due to smear-biopsy incompatibility or persistent low-grade dysplasia were excluded. Patients with a negative HPV test and an unknown HPV genotype were also excluded. Among the patients scheduled for excision (n=404), 74.5% had a s-HPV and 25.5% had a m-HPV infection. The proportion of CIN 1, 2 and 3 per patient in the m-HPV group was significantly higher than the s-HPV group (p=0.017). When this analysis was made for the number of CIN 2+3 per patient in the s-HPV and m-HPV groups, it was 1.29 (389/301) and 1.36 (140/103), respectively, and no difference was found (p=0.491). Patients in the m-HPV group, who underwent more colposcopic cervical biopsies, had higher numbers of CIN lesions, regardless of age and cytology results.