Adhemar Longatto-Filho

Comparative performance of cobas 4800 HPV Test and Anyplex II HPV HR for high-risk human papillomavirus detection

ABSTRACT Numerous molecular tests are available to detect human papillomavirus (HPV). We compared the analytical performance of cobas and Anyplex for detection of high-risk (HR) carcinogenic HPV genotypes, assessed the composition of HPV types (other than 16 and 18) that influenced cobas performance, and considered the impact of viral load on test performance. We used data from the Early Detection of Cervical Cancer in Hard-to-Reach Populations of Women Through Portable and Point-of-Care HPV Testing project, which involved collection (2019–2022) of cervicovaginal samples from 1,042 women aged 21–74 years in Belgium ( n = 244), Portugal ( n = 309), Brazil ( n = 244), and Ecuador ( n = 245). Samples were tested by cobas (provides individual results for HPV16 and HPV18 and a pooled result for 12 other HR-HPV types) and Anyplex (provides separate results for 14 HR-HPVs). We calculated HPV positivity by each test and compared performance between tests by calculating Cohen’s kappa statistics. Based on 938 samples with complete data from both tests, positivity rates by cobas were 13.4%, 3.6%, 34.3%, and 45.3% for HPV16, HPV18, 12 pooled HR-HPVs, and any HR-HPV, respectively. Corresponding HPV positivity rates by Anyplex were 14.9%, 3.7%, 37.9%, and 50.0% for the same categories, respectively, with high concordance; kappa statistics were 0.90, 0.87, 0.82, and 0.85, respectively. Based on 355 samples that tested positive for at least 1 of the 12 pooled HR-HPVs, most types showed high agreement (80.9%–100.0%) between individual-Anyplex and pooled-cobas HPV results, except for HPV68 (61.3% agreement). Our findings suggest that the two commercial tests may have different performances, depending on the specific HPV types detected, emphasizing the need for continued research on conditions that may affect these tests, especially for less common or less studied HPV types. IMPORTANCE This study compared two commercial tests—cobas and Anyplex—for detecting high-risk HPV types in women undergoing routine cervical cancer screening or referred for colposcopy. Both tests provide separate results for HPV16 and HPV18, but Anyplex also identifies the remaining 12 high-risk HPV types individually, while cobas groups them together. Overall, we found a high level of agreement between the two tests, supporting their use in clinical practice. However, differences in detecting certain HPV types, particularly those that are less common or less studied, emphasize the importance of choosing the right test. As more countries switch to HPV-based cervical cancer screening, using tests that provide detailed results could help improve risk assessment and optimize patient care.

Critical Evaluation of Pap Test Adherence to Routine Screening in Amazonas State, Brazil

Introduction: This study critically evaluates adherence to Pap test screening practices in cytology-based cervical cancer screening in the state of Amazonas over a 10-year period. Materials and Methods: A retrospective analysis was conducted of the results of cytological screening examinations (Pap test) in Amazonas State from 2013 to 2023. For this purpose, Brazilian public databases Cervical Cancer Information System (SISCOLO) and Cancer Information System (SISCAN) (from the Department of Information and IT of the Unified Health System [DATASUS]) were consulted. Results: There was a decrease in the number of Pap tests performed during the period from 2019 to 2021, likely related to the COVID-19 pandemic. This was followed by a subsequent increase in the post-pandemic period. Notably, in municipalities with fewer than 10,000 annual Pap examinations there was a decrease in the average number of tests when comparing the years 2016–2018 to 2013–2015, and an even greater decrease during the pandemic. Conclusions: There is considerable variation in utilization of the cytological Pap test across different municipalities. This lack of uniformity throughout the state likely compromises the capacity to detect early stage cervical intraepithelial lesions.

Evaluation of Elafin Immunohistochemical Expression as Marker of Cervical Cancer Severity

<b><i>Introduction:</i></b> The main risk factor for the development of cervical cancer (CC) is persistent infection by human papillomavirus (HPV) oncogenic types. In order to persist, HPV exhibits a plethora of immune evasion mechanisms. PI3/Elafin (Peptidase Inhibitor 3) is an endogenous serine protease inhibitor involved in epithelial protection against pathogens. PI3/Elafin’s role in CC is still poorly understood. <b><i>Materials and Methods:</i></b> In the present study, we addressed PI3/Elafin protein detection in 123 CC samples by immunohistochemistry and mRNA expression in several datasets available at Gene Expression Omnibus and The Cancer Genome Atlas platforms. <b><i>Results:</i></b> We observed that PI3/Elafin is consistently downregulated in CC samples when compared to normal tissue. Most of PI3/Elafin-positive samples exhibited this protein at the plasma membrane. Besides, high PI3/Elafin expression at the cellular membrane was more frequent in in situ stages I + II than in invasive cervical tumor stages III + IV. This indicates that PI3/Elafin expression is gradually lost during the CC progression. Of note, advanced stages of CC were more frequently associated with a more intense PI3/Elafin reaction in the nuclei and cytoplasm. <b><i>Conclusion:</i></b> Our results suggest that PI3/Elafin levels and subcellular localization may be used as a biomarker for CC severity.

Self-Sampling as a Plausible Alternative to Screen Cervical Cancer Precursor Lesions in a Population with Low Adherence to Screening: A Systematic Review

<b><i>Background:</i></b> Cervical cancer is the fourth most common cancer in the world. A raised incidence and mortality parallel a low participation rate in screening, namely screening in poor countries and among specific populations of developed countries. Cervical or vaginal self-sampling may increase adherence to screening due to its low costs and elevated sensibility and specificity. Our main goal was to compare self-sampling with physician sampling and to evaluate the participation rate of women formerly non-adherent to cervical cancer screening. <b><i>Study Design:</i></b> We identified relevant studies from PubMed. Studies were eligible for inclusion if they fulfilled the following criteria: women aged between 16 and 80 years, poor and developed countries that use self-sampling, women with a low participation rate in screening and studies published since 2013. After fully reading the articles, data were extracted to an Excel sheet to ­display all relevant information in an organized manner. ­<b><i>Results:</i></b> We identified 18 studies, which altogether enrolled 22,118 women. Self-sampling and physician sampling are quite similar regarding the HPV detection rate. Women have a low participation rate mainly because of limited access to health services, religious and culture beliefs, and lack of time due to several demanding tasks in daily life. Self-sampling shows a significant increase in acceptability and preference compared to physician sampling. <b><i>Conclusion:</i></b> Self-sampling is a reliable method to involve women in opportunistic or organized screening programs for cervical cancer prevention.

Combined Oral Contraceptive Use and the Risk of Cervical Cancer: Literature Review

AbstractCervical cancer (CC) is caused by persistent infection of human papillomavirus of high oncogenic risk (hr-HPV); however, several cofactors are important in its carcinogenesis, such as smoking, multiparity, and prolonged use of oral hormonal contraceptives (COCs). Worldwide, 16% of women use COCs, whereas in Brazil this rate is of ∼ 30%. The safety and adverse effects of COCs are widely discussed in the literature, including the increase in carcinogenic risk. Due to the existence of several drugs, combinations, and dosages of COCs, it is hard to have uniform information in epidemiological studies. Our objective was to perform a narrative review on the role of COCs use in the carcinogenesis of cervical cancer. Several populational studies have suggested an increase in the incidence of cervical cancer for those who have used COCs for > 5 years, but other available studies reach controversial and contradictory results regarding the action of COCs in the development of CC.

Self-sampling for HPV genotyping: a study of vaginal and urine collection in Brazilian women with high-grade lesions

In Brazil - a country of continental dimensions with marked socioeconomic disparities - the use of self-collected samples and first-void urine for cervical cancer screening may be particularly valuable. This study aimed to assess the acceptability of two self-sampling approaches - first-void urine collection and vaginal self-sampling - among women diagnosed with high-grade cervical lesions (CIN2+) referred to a tertiary care center. Additionally, the study evaluated the concordance of high-risk HPV (hrHPV) test results obtained from self-collected samples compared to those collected by a healthcare professional. This cross-sectional study included 100 women. Participants completed a structured questionnaire on clinical history, demographics, gynecological and obstetric background. Following an instructional video, they performed self-collection of urine and vaginal samples. All participants then underwent colposcopic examination for lesion assessment and therapeutic planning. HPV DNA testing was conducted, and agreement analysis was performed between sample types. Both urine and vaginal self-collection methods were reported as easy and comfortable. Instructions were considered easy or very easy by nearly all participants for all collection methods. Clinician-collected sampling was associated with higher embarrassment and discomfort. Agreement analysis showed excellent concordance for HPV 16 and other high-risk HPV types between self-collected, urine, and clinician-collected samples, with all comparisons reaching statistical significance. Urine and vaginal self-collection are feasible, acceptable, and reliable methods. Urine sampling was the preferred method in the present study. High concordance with clinician-collected samples confirms their clinical utility, and the positive response to instructional videos highlights the importance of educational support.

Gastrin-releasing peptide receptor: a promising new biomarker to identify cervical precursor lesions and cancer

This study aimed to verify the relation between gastrin-releasing peptide receptor (GRPR), oncogenic Human Papillomavirus (HPV) and cervical lesions severity. GRPR mRNA levels were evaluated in cervical cancer-derived cell lines and in primary keratinocytes expressing HPV16 oncogenes by RT-PCR. GRPR protein expression was assessed by immunohistochemistry in organotypic cell cultures derived from keratinocytes transduced with HPV16 oncogenes and in 208 cervical samples, including 59 non-neoplastic tissue, 28 cervical intraepithelial neoplasia grade 3 (CIN3), 44 squamous cell carcinomas (SCC) and 77 adenocarcinomas (ADC). Generic primers (GP5+/GP6+) were used to identify HPV infection in tissue samples. Experiments involving cell lines were analyzed through non-parametric tests (Kruskal Wallis), and Fisher's Exact Test for human tissues samples. All statistical tests were considered significant at p <0.05. Immunohistochemical evaluation was conducted independently and blindly by two observers (AD- LO). Any discordant findings were resolved through discussion to reach a consensus score. GRPR mRNA levels were not increased in cells expressing HPV16 or HPV18 oncogenes. However, at the protein level, GRPR was upregulated in organotypic cell cultures containing HPV oncogenes. Besides, it was identified an association between GRPR expression and cervical lesion severity (p < 0.0001). The detection rate of high-risk HPV DNA was directly correlated with cervical disease. Nonetheless, HPV infection was not directly associated with GRPR in cervical samples. GRPR expression is highly predictive of cervical lesion severity, irrespective of HPV infection and might contribute to improving patient's therapeutic management as well as being used a marker of disease progression.

Ongoing Challenges in Maintaining the Diagnostic Quality of Cervical Cytopathology: Is It Possible to Minimize Differences in Morphological Interpretations?

Background: The efficiency of the cytological test largely depends on the control and quality assurance of laboratory procedures, to avoid false results. The objectives of this descriptive review were to point out and discuss the main mechanisms for controlling and ensuring the quality of cytological diagnosis. Summary: A critical revision was performed to identify the principal challenges involved in the processes of all the main issues related to the morphological alterations that characterize a true-related lesion in the daily routine of the Pap test examination. Principles of QC and QA are already being implemented worldwide, and the positive aspects and limitations of these are discussed as well as proposing alternatives when pertinent. Most of the articles evaluated highlighted the necessity of implementing audit mechanisms to control the performance of the professionals involved with the cytology evaluation. Key Message: Promising data from the new image-based technological arsenal appear to be a remarkable tool for improving cytological evaluation, reducing errors of interpretation and serving as a powerful arm for cytology teaching.

A critical evaluation of the status of HPV vaccination in São Paulo State, Brazil

Cervical cancer, whose well-recognized etiological agent is the Human Papillomavirus (HPV), is responsible for approximately 300,000 deaths worldwide, 80% of cases occurring in developing countries. In Brazil, 17,010 cases were expected, with 2,550 cases in São Paulo State, in 2023. The Papanicolaou test is the diagnostic method for the detection of precancerous lesions of the cervix, and HPV vaccination is now available for prevention. This is a descriptive and exploratory, retrospective investigation, carried out through analysis of data obtained from Brazilian Information Technology (DATASUS) of the Brazilian Federal Government: Cancer Information System (SISCAN), Brazilian National Immunization Program (PNI) and Mortality Information System (SIM). Electronic Library and data made available by the Government of the State of São Paulo. The number of women in São Paulo State who underwent cytological examinations and histological tests for cervical cancer decreased between the Years 2013 and 2022. The continuous increase in cervical cancer over the study period was probably due to the lack of adherence to the primary and secondary prevention opportunities offered by the Public Health Authorities.