Pap Test Research Articles

Multi-institutional 10-year retrospective review of amoeba diagnosed on cytologic evaluation of anal pap tests: what is the significance?

Intestinal amoebae are usually transmitted via ingestion of amoebic cysts in fecally contaminated water or food. However, other modes of transmission include sexual contact through anal-oral sex. While the primary role of anal cytology is the detection of anal cancer and precursor lesions, organisms can also be identified. Despite this, assessment of the clinical significance of cytologic identification of amoebae is lacking in the literature. A 10-year retrospective review of the pathology archives of 2 institutions in Boston, Massachusetts was conducted. Anal Pap tests that identified amoeba were reviewed and correlated with the medical records for investigation into clinical parameters and patient management. A total of 46 cases were identified between the 2 sites. The majority of patients were male (95.7%) and endorsed having sex with men (84.8%). Only a minority endorsed recent travel (6.5%). Most of the patients were also HIV (human immunodeficiency virus)-positive (71.1%) with all of these patients being well-controlled on antiretroviral therapy. Most patients were asymptomatic (87.0%). On review of the anal Pap tests, the average organism number per case was 35.4. In the majority of cases, follow-up microbiology testing for confirmation and/or speciation was not performed (89.1%) and were not treated (93.5%). While identification of amoeba is possible on anal cytology, the clinical significance remains unclear as most patients were asymptomatic and not treated in this series. Ultimately, the clinical setting likely plays an important role in determination of management.

Sexual and reproductive health cancer screening avoidance: The role of body image

The current study examines the relationship of female genital self-image, weight-related cancer screening avoidance, and internalized weight stigma among cisgender women that can provide knowledge about avoidance of life-saving preventative healthcare. This cross-sectional survey included a convenience sample of 384 U.S. cisgender women who were 18+. The sample was primarily white (n = 260, 67.7 %) with a mean age of 33.18 years. 28.4 % reported avoiding a pap smear, 27.1 % avoided a clinical breast exam, and 29.4 % avoided a mammogram. Using multivariate logistic regressions, our results show high internalized weight stigma moderates the relationship of positive genital self-image on weight-related genital and breast cancer screening avoidance. Thus, the odds of avoiding screenings are positive, where the odds of avoidance slightly decreases from the interaction term as female genital body image increases. Interventions to improve female genital body image among cisgender women may lessen the effects of internalized weight stigma on avoiding reproductive cancer screenings. BMI was only a predictor for avoiding pap tests. Further examination is warranted because BMI and sexual health behaviors are not usually associated in body image research. Clinical workforce training is needed to educate providers about the harm of weight stigma and its relationship with healthcare avoidance.

Navigating Practice Issues Related to the Unsatisfactory Cervicovaginal Papanicolaou Test: Survey Results of Laboratories Participating in the 2020 College of American Pathologists PAP Education Program.

Unsatisfactory Papanicolaou (Pap) tests pose a unique set of challenges to the laboratory with regard to their processing, review, reporting, and performance of human papillomavirus (HPV) testing. There are no standardized guidelines for the review process and handling of unsatisfactory Pap tests. To assess the current practice patterns regarding various aspects of the unsatisfactory Pap test, from processing to reporting, across laboratories worldwide. A supplemental questionnaire was mailed to laboratories participating in the 2020 College of American Pathologists (CAP) Gynecologic Cytopathology (PAP Education) Program, requesting data regarding the unsatisfactory Pap test. Of 1520 participating laboratories, 619 (40.7%) responded, and the responses of 577 laboratories were included for further analysis. Only 64.6% (373 of 577) laboratories used the unsatisfactory Pap test criteria as specified by the 2014 Bethesda System. About three-quarters of the respondents (433 of 576; 75.2%) routinely rescreened unsatisfactory Pap tests. Routine repreparation of such Pap tests was performed by 54.9% (316 of 576) of laboratories, and 52.0% (293 of 563) used glacial acetic acid for repreparing excessively bloody specimens. HPV test results were reported for unsatisfactory Pap tests, always or sometimes, by 62.4% (353 of 566) of respondents. This CAP survey reveals important information regarding the practice patterns pertaining to several aspects of the unsatisfactory Pap test. It also provides valuable insight into the quality assurance measures that can be implemented for such tests. Future studies can further aid in the standardization of all components of the handling of unsatisfactory Pap tests for overall quality improvement.

A Cervical Cancer Screening Toolkit for Somali Women: A Pilot Randomized Controlled Trial.

This study aims to determine whether a novel cervical cancer screening toolkit will increase rates of pap test completion and HPV vaccination among Somali women living in the United States. We conducted a randomized controlled, pilot trial from June 2021 to February 2022. Somali women aged 21 to 70 were randomized to either receive a toolkit (infographic, video and an in-person health seminar) or not. Health passports confirming a completed pap test and/or HPV vaccination by clinician signature were used to measure outcomes. The primary outcome was pap test completion and the secondary outcome was HPV vaccination. We enrolled 57 participants. Patients randomized to the treatment arm were significantly more likely to have had a pap test (53.7% vs. 3.7%, p < 0.0001) and were also more likely to have received the HPV vaccine (10.7% vs. 3.7%, p = 0.6110). This toolkit increased rates of pap test completion and more participants in the intervention arm received HPV vaccination, though numbers were low. The study design may serve as a reproducible model to determine the effectiveness of patient education materials.

Factores socioculturales que influyen en las usuarias de 50 años para la toma de papanicolau de un hospital nasca

Objective: The objective of this study was to determine the sociocultural factors in 50-year-old users for Pap smears at the Ricardo Cruzado Rivarola de Nasca hospital during the period December 2021 - February 2022. Material and Methods: Observational, analytical, and prospective study. The study population is made up of 80 50-year-old users who attend the Ricardo Cruzado Rivarola hospital for Pap tests at the gynecology-obstetrics office and the sample is 67 users, selected by non-probabilistic sampling for convenience. Results: For the first analysis, in simple and multiple regression, it was found that women with high levels of knowledge had a 384% and 382% higher frequency of having ever undergone a PAP test compared to those with low levels of knowledge. Conclusions: The level of knowledge about the PAP has a relevant influence on having carried out a PAP; but also the women who had a high level were the ones who had the highest frequency of having done it. This added to the number of children; possibly because in the circles of women with children one of the topics to be discussed is about maternal health and within them the PAP.

The detrimental consequences of two consecutive disasters impacting cytopathology in Ukraine: COVID followed by thewar.

The Covid-19 pandemic and ongoing war in Ukraine caused unprecedented disruption in healthcare, including cytopathology activities. This paper elucidates the effect of two consecutive disasters-the COVID-19 pandemic followed by the war-on cytopathology practice in Ukraine through a single-centre retrospective study. Total testing volumes, geographic distribution, and indicators of laboratory operations were assessed during three periods of 3 months each: the first 3 months of the acute phase of the war (March-May 2022, period 1); summer (June-August 2022, period 2); and the fall (September-November 2022, period 3, associated with massive attacks on the energy infrastructure in Ukraine). These data were compared with the corresponding periods in 2020, during the COVID-19 pandemic, and in 2021, the post-lockdown period. The ongoing war in Ukraine has caused a dramatic disruption in routine health maintenance and cytological practice. A net decline in both PAP testing and non-gynaecological pathology was associated with a geographic redistribution of cytopathological testing, and an increase in the rate of abnormal sample reporting. Despite these challenges, cytopathology practice in Ukraine demonstrates resilience, allowing for maintaining the healthcare system and addressing the needs of the civil population during the war. The ongoing war in Ukraine heavily affected cytological practice. The decline in PAP testing during the early period of the war was associated with an increase in the abnormal sample rate. Further study of the war's impact on the cervical pathology rate and the health of the population in the next decades is needed.

HPV Cotesting of Unsatisfactory Papanicolaou Tests: Implications for Follow-up Intervals.

The 2019 American Society of Colposcopy and Cervical Pathology management guidelines recommend that patients with an unsatisfactory Papanicolaou (Pap) test (UPT) and negative human papillomavirus (HPV) cotest undergo repeat age-based screening in 2 to 4 months. The rationale is that a negative HPV test in the setting of an UPT may reflect an inadequate sample and therefore should not be interpreted as truly "negative." For patients 25 years and older who are cotested, if HPV is positive for the 16 or 18 genotypes, direct referral for colposcopy is recommended. Our study aimed to determine if a negative HPV cotest result is predictive of the absence of a high-grade squamous intraepithelial lesion (HSIL) and whether these patients may be called back for repeat testing at an interval longer than 2 to 4 months. Follow-up cervical cytology and biopsy results in women with UPT and HPV cotests from January 2017 to December 2021 were collected. Original UPT and HPV cotest results were correlated with the follow-up Pap and biopsy results. There were 1,496 (2.28%) UPT cases out of 65,641 total Pap tests. Among the 1,496 UPT cases, 1,010 (67.5%) had HPV cotesting; 676 (45.1%) were followed by repeat Pap or biopsy within 4 months and 850 (56.8%) within 12 months. The total follow-up rate was 81%, with a range of 3 days to 36 months. The HSIL rate in HPV-positive cases was 5.7% (3/53) vs 0.4% (2/539) (P = .006) in HPV-negative cases. In UPT, HPV cotesting showed negative predictive values for low-grade and high-grade squamous intraepithelial lesion detection of 98.5% and 99.6%, respectively, while positive predictive values were 19% and 5.7%. A negative HPV cotest in individuals with UPT predicted the lack of HSIL in our study. Compliance with the recommended follow-up time of 2 to 4 months for women with UPT was low (45.1%). Our study suggests that women with UPT and negative HPV cotest may be safely called back at an interval longer than 4 months.

Population-level use of gynecological health services by female youth with intellectual/developmental disabilities in British Columbia Canada

BackgroundThere is evidence that female youth with intellectual/developmental disabilities (IDD) experience poorer gynecological care compared to female youth without disabilities. ObjectiveThe objective of this study was to obtain baseline data on visits to a health care provider for a gynecological issue for females with IDD and compare that information to the experiences of female youth without IDD. MethodsThis study is a retrospective cohort study using population-level administrative health data from 2010 to 2019 for females aged 15–24 years, with and without IDD. Results6452 female youth with IDD and 637,627 female youth without IDD were identified in the data. Over the ten-year period, 53.77% of youth with IDD and 53.68% of youth without IDD had a visit to a physician for a gynecological issue. However, as females with IDD aged, the number of people seeing a physician for a gynecological issue decreased. In the group aged 20–24 years, 15.25% of females with IDD and 24.47% of females without IDD (p < 0.0001) had a Pap test done at any time; 25.94% of females with IDD had a visit for contraception management and 28.38% of females (p < 0.0001) without IDD had a visit for contraception management. Gynecological care also varied by type of IDD. ConclusionsFemales with IDD had a similar number of visits for a gynecological issue as female youth without IDD. However, the reasons for visits and the age at which visits occurred differed between youth with and without IDD. As females with IDD transition into adulthood, gynecological care must be maintained and improved.

Knowledge and practices about cervical cancer and human papilloma virus among the women and the influencing factors

ABSTRACT Knowledge about Human Papilloma Virus (HPV), timely screening programs and early vaccination are crucial to prevent cervical cancer. Existing studies have reported a lack of knowledge and awareness about HPV, Pap tests and cervical cancers. This study aims to evaluate the knowledge, awareness and behaviors of the women about cervical cancer, HPV and HPV vaccine. This descriptive, cross-sectional design study was carried out at a family health center in Istanbul with the participation of 1050 women from March to July 2021. Information form and HPV Knowledge Scale (HPV-KS) were used for data collection. Descriptive statistics and multiple regression analysis were used for data analysis. The mean score and standard deviation obtained from the HPV-KS were 11.80 ± 8.02. Although 77.5% of the participants had heard about Pap test, 57.9% did not have Pap test before. Besides, although 55.2% and 60.4% of the participants had heard about cervical cancer and HPV vaccine, respectively, only 6.6% had HPV vaccine before. Multiple regression analysis showed that the sociodemographic and clinical variables accounted for 22.7% of the knowledge about HPV (p < 0.001). The participants had low levels of knowledge about cervical cancer and HPV infection and vaccination. The study results can be used to inform educational interventions, which in turn, can increase knowledge in screening recipients.

Natural history of false-negative Papanicolaou smears: A prospective study using screening colposcopy in addition to cytology.

The regular Papanicolaou (Pap) smear is a cornerstone of women's preventive healthcare. The introduction of the regular Pap smear as a screening tool for cervical cancer has markedly decreased the number of deaths from cervical cancer. During the past decade, however, the rate of death from cervical cancer has remained relatively static. This screening method is known to have a high rate of false-negative results; therefore, serial examinations are necessary for optimal sensitivity. The sensitivity of the routine pelvic examination is further increased with the addition of colposcopy to cytologic screening. Patients identified as having disease by colposcopy and biopsy but not by Pap smear (false-negative), are followed up, and the progression of their disease is documented until the Pap smear reflects the known change. A total of 276 patients whose Pap tests showed "no abnormal cells" and mild dysplasia (CIN-1 [cervical intraepithelial neoplasia]) by colposcopy and biopsy, who opted not to be treated, were followed up as a function of time. The results of the Pap test on the 276 patients progressed to a low-grade squamous intraepithelial lesion (LGSIL) or higher within 42 months. In this study, false-negative Pap tests that were identified by simultaneously performed colposcopy became positive within 42 months; therefore, careful repetitive screening is necessary. The author argues that a definitive, prospective study on the addition of colposcopy or similar adjunct procedure to the routine pelvic examination, in selected cases, is needed.

A global perspective on cervical cancer screening: a literature review

Cervical cancer is a significant health issue for women globally, with over 600,000 new cases and 342,000 deaths in 2020. Effective cervical screening programs, including primary screening methods such as Pap tests, human papillomaviruses (HPV) deoxyribonucleic acid (DNA) testing, and liquid-based cytology, can prevent and detect early cancer cases, improving morbidity and mortality rates. However, screening uptake varies widely across countries, with high-income countries have achieved over 70% uptake but low- and middle-income countries lagging due to a lack of recommendations and centralised screening systems. Even in high-income countries in the Middle East, uptake remains low, requiring further investigation. This review explores barriers to screening programme delivery and highlights centralised and multi-level interventions that have proven effective worldwide. Future research should focus on culturally sensitive approaches, healthcare infrastructure strengthening, and community education and outreach programs to address uptake disparities and barriers.

Diagnostic significance of GATA 3, TTF-1, PTH, chromogranin expressions in parathyroid fine needle aspirations via immuonocytochemical method.

Additional studies may be needed when evaluating parathyroid fine needle aspirates (FNA) due to cytomorphologic similarities. The liquid-based cytology (LBC) is an effective cytology method which provides additional slides for ancillary techniques. We aimed to examine the immunocytochemical (ICC) GATA 3, TTF-1, which chromogranin and PTH expressions and to determine their diagnostic importance in parathyroid FNAs which were prepared via LBC technique. In total, 45 parathyroid FNA cases were examined in our laboratory for 12-month period. Thirty five cases without cell blocks were included in the study. Ten cases were excluded from study (five cases were nondiagnostic, five cases had cell blocks). For main LBC cytology slides SurePath pap test kit was used. For ICC study 4 PAP-stained LBC slides were obtained from each case's residual material. The slides were then stained with GATA 3, TTF-1, PTH and chromogranin. Of 35 cases, 8 were male and 27 were female. The mean age of the cases was 52.25 ± 11.44 (range 27-72). The mean diameter of the lesions was 18.54 ± 7.66 mm (range 6-32 mm).The most used antibody was TTF-1 (in all 35 cases). The positivity rates were 84.8%, 83.9% and 81.8% for GATA 3, PTH and chromogranin, respectively. TTF-1 was completely negative in 100% of cases. All patients had high PTH washout values (29 patients, PTH washout value was >5000 pg/mL). In 30 cases, histopathologic follow-up was available. The cyto-histological correlation was 100%. Based on the PTH washout values for cases without resection, the diagnostic accuracy of FNA was determined as 100% in detecting the parathyroid origin. Among the antibodies, the most specific and sensitive antibody was TTF-1 (100%). The sensitivity values of the antibodies indicating parathyroid origin for GATA3, PTH and chromogranin were 86.67%, 83.33% and 81.82%, respectively. The addition of ICC studies applied to LBC slides is also very helpful in diagnosis, especially in FNAs with limited material such as parathyroid. GATA3 is a more reliable ICC marker determining parathyroid origin. In differentiation from thyroid origin, the addition of TTF-1 to the ICC panel increases the diagnostic accuracy to 96.67%. Although PTH and chromogranin have lower sensitivity, they are still reliable markers to detect parathyroid origin.

The possibilities of adapting the re-processing protocol in the practice of the ThinPrep Pap test usage

The method of liquid cytology is a newer diagnostic test compared to the traditional cytological examination of scrapings from the cervix and allows one to standardize the morphological examination and reduce the number of unsatisfactory samples. One of the factors determining the adequacy of the Pap test is the number of squamous epithelial cells in a sample. The abundance of blood elements, mucus, lubricant, or inflammatory elements may affect the informativeness of the sample. In the practice of the ThinPrep Pap test usage, when receiving an inadequate sample, a protocol for re-processing the contents of the vial is used, in order to neutralize the possible influence of such impurities on the final result. Here, we present a review on the methods of the re-processing protocol application and our own experience of adapting the protocol in the practice of a clinical diagnostic laboratory.

Effectiveness of family-centred sexual health education and HPV self-sampling in promoting cervical cancer screening among hard-to-reach indian women in rural and tribal areas: a community-based pilot study

BackgroundWhile cervical cancer deaths have declined steeply in high-income countries due to the widespread use of the Papanicolaou test (Pap test), the same trend has not emerged in low or middle-income countries (LMICs). Access to screening in LMICs like India is limited due to barriers such as limited healthcare infrastructures, lack of sexual health education, and stigma demarcating sexually transmitted infections (STIs). HPV self-sampling (HPV-SS), a woman–centered and at-home method for screening, can be utilized as a unique screening tool to overcome some of these barriers. Our study examined the effectiveness of HPV-SS, supported by family-centred arts-based sexual health literacy on the uptake of cervical cancer screening among hard-to-reach women in rural and remote areas in India.MethodsOur community-based mixed methods pilot study recruited 240 participants (120 women and 120 male partners or family members) through female Accredited Social Health Activists (ASHA) across 3 Indian villages of Shirgoan, Khodala, and Jamsar in Palghar district. Inclusion criteria included women ages 30–69 who were under or never screened (UNS) and their male partners/family members aged 18 or over. Knowledge and attitudes about cervical cancer and screening and their perceived stigma surrounding STI were assessed using validated scales prior to and after attending a 2-hour arts-based sexual health education (SHE). In addition, participants’ uptake of cervical cancer screening was assessed after attendance in SHE.FindingsResults revealed significant improvement in knowledge and attitudes about cervical cancer and screening, and a reduction in the STI stigma after participation in SHE sessions (overall mean difference in Knowledge: z = 6.1 ± 2.4, P < 0.001; attitudes about Pap-test and VIA: z = 2.2 ± 8.4, P < 0.001 and z = 2.9 ± 8.2, P < 0.001; STI stigma: z = 2.8 ± 12.4, P < 0.001). 118 out of 120 female participants chose to be screened and 115 opted for HPV-SS.ConclusionsThe implementation of HPV-SS coupled with family-centered arts-based and culturally appropriate SHE is highly promising in promoting cervical cancer screening among hard-to-reach women. Evidence from our study can be used to advance public health policies and inform the scale-up of similar initiatives in other villages and states across rural India and other LMICs.

Changes in the Frequency and Type of Barriers to Reproductive Health Care Between 2017 and 2021

Previous research has documented individual-level barriers to reproductive health services, but few studies have examined national trends. To determine whether the number and type of barriers to reproductive health care experienced by US women of reproductive age changed from 2017 to 2021. This cross-sectional study used serial survey data, weighted to be nationally representative, collected in August 2017 and December 2021 from members of Ipsos's KnowledgePanel who were aged 18 to 49 years and assigned female at birth. Having experienced barriers to reproductive health care over the past 3 years. The primary outcome was number and type of barriers to reproductive health care services, including Papanicolaou tests or birth control, experienced in the past 3 years. Increases in barriers to reproductive health care were measured using multivariable linear regressions adjusted for age, race and ethnicity, education level, employment status, metropolitan area, geographic region, household income, and language (English vs Spanish). Of 29 496 KnowledgePanel members invited, 7022 (mean [SD] age, 33.9 [9.0] years) and 6841 (mean [SD] age, 34.2 [8.6] years) completed the 2017 and 2021 surveys (50% and 45% response rates, respectively). Among 12 351 participants who indicated that they had ever tried accessing reproductive health services, 35.9% (95% CI, 34.8%-37.0%) were aged 30 to 39 years; 5.5% (95% CI, 4.9%-6.2%) were Asian or Pacific Islander, 13.7% (95% CI, 12.8%-14.6%) were Black, 19.1% (95% CI, 18.1%-20.1%) were Hispanic, 58.2% (95% CI, 57.0%-59.5%) were White, and 3.5% (95% CI, 3.1%-4.0%) were multiracial or of other race or ethnicity; and 11.7% (95% CI, 11.0%-12.5%) were living below 100% of the federal poverty level. Covariate distribution was similar across years. In bivariable analyses, participants were more likely to report experiencing a given barrier in the past 3 years in 2021 than in 2017 for all but 2 barriers. More people experienced 3 or more barriers in 2021 (18.6%; 95% CI, 17.3%-20.0%) than in 2017 (16.1%; 95% CI, 14.9%-17.4%) (P = .008). In multivariable analyses, the mean number of barriers increased significantly from 1.09 (95% CI, 1.02-1.14) in 2017 to 1.29 (95% CI, 1.22-1.37) (P < .001) in 2021. Participants who were aged 25 to 29 years (0.42; 95% CI, 0.37-0.47), identified as Hispanic (0.41; 95% CI, 0.38-0.45), had no high school diploma or General Educational Development test (0.62; 95% CI, 0.53-0.72), lived below 100% of the federal poverty level (0.65; 95% CI, 0.55-0.73), and took the survey in Spanish (0.87; 95% CI, 0.73-1.01) saw the greatest increases in mean number of barriers between 2017 and 2021. The study findings suggest that barriers to reproductive health care increased between 2017 and 2021, with the largest increases observed among individuals from historically disadvantaged populations. Efforts are needed to ensure that reproductive health care access remains a priority.

Gynecological Care Among Brazilian Women Who Have Sex with Women: A Respondent-Driven Sampling Study.

Purpose: This study aimed to describe the gynecological care provided to Brazilian women who have sex with women (WSW). Methods: Respondent-driven sampling was used to recruit Brazilian WSW. The survey questions, concerning gynecological care, were designed in Portuguese by medical professionals, medical students, and LGBTQIA+ community members, including the authors. The statistical analyses were weighted to account for the likelihood of recruitment. Results: From January to August of 2018, 299 participants were recruited in 14 recruitment waves. The mean age of the WSW was 25.3 years. Most (54.9%) identified as lesbian and had been involved in past-year sexual intercourse mainly with cisgender women (86.1%). The WSW also reported having sex with cisgender men (22.2%), transgender men (5.3%), nonbinary people (2.3%), and transgender women (5.3%) in the last year. More than a quarter of the WSW did not have regular appointments with a gynecologist: 8.0% (95% confidence interval [CI] = 4.2-11.6) and 19% (95% CI = 12.8-25.2) stated that they had never gone to the gynecologist or they had only gone for emergencies, respectively. Almost one-third had never had cervical cancer screening (cervical cytology, Pap test or Pap smear). Most women justified avoiding the test because they felt healthy, thought it would hurt, or feared a health professional might mistreat them. Conclusion: Gynecologists should avoid heteronormative assumptions, inquire about sexual practices, orientation, and identity separately, and provide Pap tests as advised to WSW.

Abstract 1978: Community Genomic Health Promotion Program evaluates precision DNA methylation biomarkers for use in self-testing and collection kits in urine, to increase cervical cancer screening rates among Latina women

Abstract Cervical cancer incidence is 32% higher among Latina women in the United States (US) and 78% higher among women in Puerto Rico compared to non-Latina White (NLW) women, yet it is largely preventable through screening. Latina women with public insurance or uninsured are significantly less likely to have had a Pap test, undergo fewer Pap and high-Risk Human Papilloma Virus (HPV) testing than NLW and Black women, and are less likely than women of other races/ethnicities to return for recommended follow-up after an abnormal Pap test. Utilizing the PRECEDE-PROCEED model, Community Based Participatory Research strategies and the NIH I-Corps hypothesis-driven research method of customer discovery, we implemented the Community Genomic Health Promotion Program in Federally Qualified Health Centers to evaluate cervical cancer screening needs in six rural municipalities in Puerto Rico. We recently validated in clinical liquid cytology samples that the CervicalMethDx test, a precision DNA methylation test, can identify HPV positive women most likely to be diagnosed with Cervical Intraepithelial Neoplasia (CIN) grades 2 and 3. We hypothesized that a molecular pathology test which identifies advanced premalignant cervical lesions in urine, can be distributed by commercial channels targeting populations with low screening rates. The CervicalMethDx quantitative Methylation Specific PCR (qMSP) test was used to quantify ZNF516, FKBP6 and INTS1 promoter DNA methylation in well-characterized, discarded, liquid cytology clinical samples from Puerto Rico. We then tested the CervicalMethDx test concordance on paired cytobrush (n=82) and urine samples (n=82) from Chile. The CervicalMethDx test correctly classified 86% of discarded liquid cytology clinical samples from HPV-positive women, when comparing DNA methylation in CIN2/CIN3 samples (n=47) to samples with a cervical pathology diagnosis of No Intraepithelial Lesions or Malignancy (n=18), with 83% Sensitivity, 94% Specificity, Area Under the Curve (AUC) of 0.88, and 98% positive predictive value (PPV). The concordance of the CervicalMethDx test in paired brush and urine samples was moderate (rho=0.45). The Wilcoxon signed-rank test showed no statistically significant differences for the CervicalMethDx test in paired brush and urine samples (p=0.14) These encouraging preliminary results show the successful identification of advanced premalignant cervical lesions in liquid cytology media and urine samples from Latina women. The data suggests that urine should be explored as a biofluid for cervical cancer screening. Future research may lead to expanded screening options, enabling women with low access to health care (public insurance or uninsured) to collect a specimen at home, and either send it to a testing facility or perform the test at home. Citation Format: Carmen Ili, Priscilla Brebi, Jaime López, Fernando T. Zamunér, Ana Purcell-Wiltz, Amanda García-Negrón, Keilyn Vale-Lassalle, Keerthana Gosala, Mirla Otero-García, Camila Rivero-Maldonado, Teresa Torres-Rivera, Adriana Pérez-Vicente, Ashley Ramos-López, David Sidransky, Mariana Brait, Rafael E. Guerrero-Preston. Community Genomic Health Promotion Program evaluates precision DNA methylation biomarkers for use in self-testing and collection kits in urine, to increase cervical cancer screening rates among Latina women [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1978.

Abstract 4179: Anal cancer risk factors and utilization of cancer preventive strategies in people living with HIV in Puerto Rico

Abstract Introduction: Anal cancer (AC) risk is greatly elevated among persons living with HIV (PLWH), particularly men who have sex with men (MSM). The incidence of anal cancer among PLWH in Puerto Rico is 60.5 per 100,000 in MSM. Smoking and sexual behaviors may enhance anal cancer risk among PLWH. Therefore, we describe the prevalence of risk factors for AC, overall and by sexual risk group (MSM, women, and men who have sex with women [MSW]). We also evaluated receipt of cancer preventive strategies: HPV vaccination and AC screening, among a sample of PLWH in Puerto Rico. Methods: A cross-sectional study was conducted (Sept 2020 - Dec 2021) among PLWH ≥26 aged in Puerto Rico (n=212). Participants answered a phone interview that collected information on relevant characteristics. AC risk factors evaluated were current smoking, age of sexual initiation (≤15 years), lifetime anal sex, 10 or more lifetime sexual partners, history of AIDS, genital warts, and HPV infection. HPV vaccination and AC screening status were also evaluated. The overall and sexual risk group specific prevalence of AC risk factors and preventive behaviors were described. In addition, the prevalence ratio (PR) of the variables of interest, comparing the sexual risk groups, was estimated with 95% confidence intervals using Poisson regression. Results: Mean age of participants was 52.8 ± (SD=10.2); 44.1% were MSM, 23.2% were MSW, and 32.7% were women. Most PLWH had public health insurance (84.4%), an education level of high-school or less (45.7%), and 9.5% had a history of intravenous drug use. Regarding the prevalence of AC risk factors, 58.3% had 10 or more lifetime sex partners, 51% had their first sexual relationship at age ≤15 years, 29.3% were current smokers, and 75.8% of women had history of anal sex. In addition, 20.8% reported being diagnosed with AIDS, 16.5% had a history of genital warts, and 12.0% had HPV infection. Only 7.5% reported HPV vaccination and 52.2% reported ever having had an anal pap test. MSM had the highest prevalence of risky sexual behaviors, although no statistical difference was seen in the PRs. For preventive strategies, women (PR: 3.13, 95% CI 1.52-6.46) and MSM (PR: 3.42, 95% CI 1.70-6.90) had higher prevalence of anal pap uptake than MSW. Conclusion: Persons living with HIV in Puerto Rico had high prevalence of current smoking use and risky sexual behavior. Despite their increased risk of AC, and only over half had been screened for AC. Continued education and awareness about AC risk factors, smoking cessation, and importance of prevention strategies are important among PLWH. Citation Format: Sandra I. Garcia-Camacho, Jeslie M. Ramos-Cartagena, Marievelisse Soto-Salgado, Vivian Colon-Lopez, Karen J. Ortiz-Ortiz, Ashish A. Deshmukh, Ana P. Ortiz. Anal cancer risk factors and utilization of cancer preventive strategies in people living with HIV in Puerto Rico. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4179.

Abstract 5596: Cell free DNA levels and fragmentation patterns in different liquid biopsy analytes (blood, urine and vaginal fluid) in cervical cancer patients

Abstract Background: Cervical cancer (CC) is the 4th most commonly diagnosed cancer among women, with approximately 528,000 new cases annually. Current screening approaches for this disease have certain limitations; Pap tests require dedicated cytopathology infrastructure, while human papillomavirus (HPV) DNA testing may lead to invasive procedures in patients with transient infection. Liquid biopsy has emerged as a minimally invasive approach to detect and monitor disease progression and treatment response. We and others have previously demonstrated the clinical utility of circulating tumor (ct)DNA to monitor HPV+ cancers. Moreover, cell free (cf)DNA fragmentation patterns have been found to be a biomarker of disease burden. This cfDNA is thought to originate largely as a result of cell death, where small fragments of ~167 bp are associated with apoptosis and larger fragments (&amp;gt;1000 bp) are associated with necrosis. Despite advances in liquid biopsy techniques, little is known about the composition of cfDNA from different analytes in CC patients. Methods: The aim of this study was to compare the presence and composition of cfDNA from different liquid biopsy analytes in patients with CC and high grade cervical intraepithelial neoplasia or dysplasia (CIN/CD). Blood, urine, and vaginal swabs were collected from 20 patients with CC and 9 patients with CIN/CD at the McGill University Health Centre. All samples were centrifuged twice to isolate supernatant. Samples were tested for HPV ctDNA by ddPCR. Analysis of fragment length was performed using the Agilent Bioanalyzer 2100. Dominant fragment was determined as the DNA fragment with the highest concentration, while overall fragment was calculated as the average fragment across all bioanalyzer peaks. Results: HPV16/18 ctDNA was detectable 14/20 CC patients and 2/6 CIN patients, and 0/3 CD patients in plasma, urine, and vaginal swab. Concordance for all sample types tested was seen in 90% of cases. On average, fragment analysis demonstrated 174, 195, and 183 bp dominant small fragments in plasma, urine and vaginal swab, respectively. Plasma samples showed only smaller fragments (range: 168-187 bp). Other analytes displayed a predominance of larger cfDNA, with an overall fragment size of 3996 and 5194 bp in urine and vaginal swab, respectively. Conclusions: HPV-DNA was detectable in all analytes sampled in patients with CC and CIN, with a trend of higher detection in CC samples. The fragmentation patterns of cfDNA varied between patients and within patients across analytes, with larger fragments - likely of necrotic origin - seen only in urine and vaginal swab cfDNA. Smaller fragments - likely related to apoptosis - were seen across all sample types studied. Overall, analysis of fragmentation may provide valuable insight into cfDNA origins in different sample types and provide a novel biomarker for diagnosis and surveillance. Citation Format: Sarah Tadhg Ferrier, Erica Mandato, Alexandra Bartolomucci, Thupten Tsering, Shuk On Annie Leung, Julia Valdemarin Burnier. Cell free DNA levels and fragmentation patterns in different liquid biopsy analytes (blood, urine and vaginal fluid) in cervical cancer patients. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5596.

Abstract 6522: Improved detection of low frequency mutations in ovarian and endometrial cancers by utilizing a highly accurate sequencing platform

Abstract Ovarian and endometrial cancers come within the top-4 for incident cancers as well as deaths in North American women. Cure rates have not improved in 30 years as high-grade subtypes continue to be diagnosed in Stage III/IV. Attempts at early diagnosis have failed because high-grade cancer cells exfoliate and metastasize while the primary cancer is small and undetectable by existing tests based on imaging and blood-based tumour markers. DOvEEgene (Detecting Ovarian and Endometrial cancers Early using genomics) is a genomic uterine pap test developed by a McGill team to screen and detect these cancers while they are confined to the gynecologic organs and curable by surgery. The test identifies pathogenic somatic mutations in uterine brush samples A high sensitivity error-reducing capture technology (DOvEEgene-SureSelectHS) utilizing duplex sequencing interrogates the exons of 23 genes involved in the development of sporadic and hereditary ovarian and endometrial cancers. We apply a combination of germline gene panel testing on saliva samples with deep duplex sequencing to detect somatic mutations at &amp;lt;0.1% VAF, interrogation of microsatellite loci for instability and low coverage WGS for copy number analysis of uterine brush samples. Currently, DOvEEgene is the only test that can discriminate ovarian and endometrial cancers in peri- and postmenopausal women from benign gynecologic diseases common in that age group. This is important because pathogenic somatic driver mutations are also associated with increasing age and benign disease. DOvEEgene incorporates a deep machine-learning derived classifier that can discriminate the mutational signature of these cancers from benign disease aiming for a sensitivity of 70% and a specificity of 100% in a population with high background mutational burden. Here we tested the Onso system, a highly accurate sequencing technology from PacBio in order to potentially increase sensitivity while driving down sequencing costs by reducing required sequencing depth vs the current NGS standard. We sequenced 15 duplex Illumina sequencing libraries produced using the DovEE assay at PE 100bp mode and compared Onso data in non- duplex sequencing mode as well as duplex sequencing mode to the original duplex sequencing method. Here, we present this comparison and highlight the benefits of high accuracy sequencing for the detection of very low frequency (&amp;lt;0.1%) somatic mutations. Citation Format: Jiannis Ragoussis, Nairi Pezeshkian, Lucy Gilbert. Improved detection of low frequency mutations in ovarian and endometrial cancers by utilizing a highly accurate sequencing platform [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6522.