Evaluation of PathTezt™, a Liquid-Based Cytology System.

Liquid-based preparation (LBP) cytology is commonly used in most laboratories these days due to its convenience and reliable results for the cervical cancer screening program. The PathTezt™ Liquid-based Pap smear is a second-generation LBP, which uses a filter-based concentration technique in processing the sample. OBJECTIVE: This study was done to evaluate the cellular fixation, morphology, quality of smear in gynae cytology, and diagnostic interpretation of cervical cytological smears produced by the PathTezt liquid-based processor. MATERIALS AND METHODS: A total of 400 pap smear samples were taken and processed using the PathTezt 2000 processor. The slides were evaluated in terms of sample adequacy, percentage of the circle covered by epithelial cells, cellular distribution, obscuring factors, and cell fixation. RESULTS: About 95.25% (381) of the samples were satisfactory for the evaluation. In 19 (4.75%) of the samples, epithelial cells covered less than 50% of the circle. A sample with good cellular distribution was seen in 92% of the cases, while 354 (88.5%) samples showed minimal inflammatory background. Almost all the smears (95.75%) had no erythrocytes in the background. All smears showed good quality fixation features toward nuclear, cytoplasm, and microorganisms. The total performance rate was 99%. CONCLUSION: Although the PathTezt liquid-based processor is still new compared to other first-generation LBP, the smears produced by this method were of high quality and it was cost-effective.

Effective reduction in inadequate Pap smears by using a saline-lubricated speculum and two glass slides.

OBJECTIVE: The aim of the study was to propose a method using saline lubrication and two glass slides to reduce the proportion of inadequate Pap smears. MATERIALS AND METHODS: This was a retrospective study of patients at a medical center in eastern Taiwan that performs 5000-6000 Pap smears annually. The extracted data only detailed the number and percentage of inadequate Pap smears. We applied two modifications to the conventional Pap smear technique. The first modification was lubricating the speculum with normal saline instead of jelly. The second modification was performing the smear on two glass slides instead of just one. We used the modified technique beginning in January 2017. Therefore, we collected data from 2016 (before the modified technique was employed) and 2018 (after the modified technique was employed). The categorical data are presented as numbers (percentages). The differences in the number and percentage of inadequate smears resulting from both techniques were assessed using the Chi-square test. RESULTS: During 2016 and 2018, 28 and 2 women received inadequate Pap smears among the total of 594 and 613 women who received Pap smears, respectively. The proportion of inadequate Pap smears was 4.71% and 0.33% in 2016 and 2018, respectively (P < 0.001). CONCLUSIONS: The use of this modified technique effectively reduced the percentage of inadequate Pap smears.

Exame citopatológico do colo uterino: descrição dos principais indicadores em um município nordestino / Cytopathologic tests of cervical: description of the main indicators in a northeastern municipality / Exámenes citopatológicos del cuello uterino: descripción de los principales indicadores en un municipio del noreste

Introdução: O câncer do colo do útero é um importante problema de saúde pública na população feminina brasileira e seu diagnóstico precoce aumenta consideravelmente a probabilidade de cura. A taxa de cura é próxima aos 100%, demonstrando que medidas preventivas, comoo exame Papanicolau, são fundamentais para prevenção desse problema. Objetivo:O objetivo deste estudo foi descrever os resultados dos principais indicadores dos exames citopatológicos em mulheres do Município de Mossoró, estado do Rio Grande do Norte no período de 2014-2019.Metodologia:Foi realizado um estudo observacional descritivo de série temporal, com levantamento retrospectivo analítico dos exames citopatológicos cadastrados no banco de dados doSistema de Informação do Câncer de Colo de Útero entre o período de 2014 e 2019. Os dados foram coletados e descritos em valores absoluto e percentual.Resultados:Um total de 46606 casos foram incluídos e analisados. Houve uma tendência de aumento na realização dos testes ao longo dos anos. 83,8% dos casosrepresentam células escamosas atípicas de significado indeterminado. 70% das lesões intraepiteliais escamosas foram diagnosticadas como sendo de baixo grau. Não houve diagnóstico citopatológico de carcinoma de células escamosas e, apenas 2 casos de adenocarcinoma in situ. Conclusões: O município de Mossoró segue um padrão nacional de maiores resultados celulares atípicos segundo os achados que surgiram. Resultados sugestivos de alto grau de malignidade existem em menor frequência nos exames preventivos e necessitam ser avaliados por exames mais invasivos (AU).

Introduction:Cervical cancer is an important public health problem in the Brazilian female population and its early diagnosis considerably increases the likelihood of a cure. The cure rate is close to 100%, demonstrating that preventive measures, such as the Papanicolau test, are fundamental for overcoming this problem. Objective:The objective of this study isdescribe the main indicators in cytopathological tests accomplished in women in the City of Mossoró, state of Rio Grande do Norte, in the period of 2014-2019. Methodology:This is a descriptive observational study of time series was carried out, in which we analyzed all citopathological tests that are registered in the System Database Information about cervical cancer between the period of 2004 and 2019 for their epidemiological profile. The data were collected and checked and demonstrated in absorbed value and percentage. Results:A total of 46606 cases were included and analyzed. There has been an increasing trend in testing in recent years. 83.8% of cases represent atypical squamous cells of undetermined importance. 70% of squamous intraepithelial lesions were diagnosed as low-grade. There was no cytopathological diagnosis of squamous cell carcinoma and only 2 cases of adenocarcinoma in situ. Conclusions:The municipality of Mossoró follows a national pattern of higher atypical cell results according tothe findings that emerged. Suggestive results of a high degree of malignancy are observed less frequently and should be evaluated by more invasive and sensitive tests for the final diagnosis (AU).

Introducción: El cáncer de cuello uterin es um importante problema de salud pública en la población femenina brasileña y su identificación precoz aumenta considerablemente la probabilidad de cura. La tasa de curación es cercana al 100%, lo que demuestra que las medidas preventivas, como la prueba de Papanicolaou, son fundamentales para prevenir ese problema. Objetivo: El objetivo de este estudio es describir los principales indicadores en las pruebas citopatológicas en mujeres del municipio de Mossoró, estado de Rio Grande do Norte, en el periodo de 2014-2019.Metodología: Este es un estudio descriptivo observacional de la serie temporal, retrospectivo en el que analizamos todas las pruebas citopatológicas que están registradas en la Información de la base de datos del sistema sobre el cáncer de cuello uterino entre el período de 2004 y 2019 para su perfil epidemiológico. Los datos fueron recopilados y verificados y demostrados en valores absolutos y porcentuales. Resultados: Se incluyeron y analizaron un total de 46606 casos. Hubo una tendencia de aumento en las pruebas en los últimos años. El 83,8% de los casos representan células escamosas atípicas de importancia indeterminada. El 70% de las lesiones escamosas intraepiteliales fueron diagnosticadas como de bajo grado. No hubo diagnóstico citopatológico de carcinoma de células escamosas y solo 2 casos de adenocarcinoma in situ. Conclusiones: El municipio de Mossoró sigue un patrón nacional de mayores resultados de células atípicas según los hallazgos que han surgido. Los resultados sugestivos de un alto grado de malignidad se observan con menos frecuencia y deben evaluarse mediante pruebas más invasivas y sensibles para el diagnóstico final (AU).

Molecular detection of oncogenic subtypes of human papillomavirus (HPV) in a group of women in the Amazon region of Brazil

The natural history of cervical cancer is strongly related to the presence of human papillomavirus (HPV) infection, with its relationship with cervical cancer being a matter of concern. It is estimated that 70% of all cervical cancers worldwide are caused by HPV 16 and 18. Accordingly, the present study aimed to contribute to the identification of HPV subtypes circulating in a group of women of Manaus-Brazil. Cervical samples were collected from 49 women, following the eligibility criteria of the study, and DNA was then extracted from the samples, which were analyzed for the presence of the virus in the genetic material through the polymerase chain reaction (PCR) using generic primers (GP05/06). Finally, identification of the viral subtypes was performed using specific primers for the detection of the main subtypes already examined (16 and 18). Positive HPV DNA was detected in 100% of the samples included in the study. Human papillomavirus 16 was the most prevalent subtype in the majority of lesions, accounting for 29 (59.2%) of the positive cases, and HPV 18 was detected in four (8.2%) women. In these 4 cases there was co-infection, with the presence of both HPV 18 and HPV 16. Therefore, 40.8% (20 cases) in which HPV DNA was detected presented infection with other subtypes of HPV not included in the study. This data has clinical implications related to cervical cancer prevention, as the current prophylactic HPV vaccines are only effective against high-risk HPV 16 and 18 subtypes.

Analysis of cervical cancer mortality rate trends in Natal-RN, Brazil, between 2000 and 2012 / Análisis de las tendencias de la tasa de mortalidad por cáncer de cuello uterino en Natal-RN, Brasil, entre 2000 y 2012

ABSTRACT Objective To describe cervical cancer mortality rates and their corresponding trends, and to analyze the spatial correlations of this type of cancer in Natal-RN, Brazil, between 2000 and 2012. Materials and Methods The simple linear regression model, the empirical Bayes method and the Global Moran's index were used for the statistical analysis. Results The mortality coefficient of cervical cancer in Natal, standardized by age range, was 5.5 per 100 000 women. All historical series for the coefficients studied were classified as stable. The Global Moran's index obtained was 0.048, with a p-value for the spatial test correlation between neighborhoods of 0.300. The average family income by neighborhood showed no significant correlation to cervical cancer mortality rates. Conclusion This study found a temporal stabilization and spatial independence trend of cervical cancer mortality rates in women from Natal, as well as the absence of correlation between these rates and the average family income of the of the participating women distributed by neighborhoods. In view of this, changes in the public policies should be made aimed at preventing the disease; adopting these measures could positively impact the screening program, improving the coverage of Pap smears and immunization campaigns against HPV, in order to reverse this trend and achieve a reduction of mortality rates.(AU)

RESUMEN Objetivo Describir las tasas de mortalidad por cáncer de cuello uterino y sus tendencias, así como analizar las correlaciones espaciales de este tipo de cáncer en Natal-RN, Brasil, entre 2000 y 2012. Materiales y Métodos Para el análisis estadístico se utilizaron el modelo de regresión lineal simple, la estimación empírica de Bayes y el índice Moran Global. Resultados La tasa de mortalidad por cáncer de cuello uterino en Natal, estandarizado por rango de edad, fue 5.5 por cada 100 000 mujeres. Todas las series históricas para los coeficientes estudiados se clasificaron como estables. El índice Moran Global obtenido fue 0.048, con un valor p de 0.300 para la correlación de prueba espacial entre vecindarios. El ingreso familiar promedio por vecindario no mostró correlación significativa con las tasas de mortalidad por cáncer de cuello uterino. Conclusión En este estudio se observó una tendencia temporal de estabilización e independencia espacial de la tasa de mortalidad por cáncer cervical en mujeres de Natal, así como la ausencia de correlación entre estas tasas y el ingreso familiar promedio de las mujeres participantes, distribuidas por vecindarios de la ciudad. En vista de esto, se sugiere que se adopten cambios en las políticas públicas dirigidas a la prevención de la enfermedad que apunten a medidas que puedan tener un impacto positivo en el programa de monitoreo, mejorando la cobertura de la prueba de Papanicolaou, así como de las campañas de vacunación contra el VPH, con el objetivo de revertir esta tendencia y lograr una reducción en las tasas de mortalidad de la enfermedad.(AU)

Human papillomavirus detection with genotyping by the cobas and Aptima assays: Significant differences in HPV 16 detection?

BACKGROUND: The primary high-risk human papillomavirus (hrHPV) assays in the United States are the cobas (Roche) and the Aptima (Hologic). The cobas assay detects hrHPV by DNA analysis while the Aptima detects messenger RNA (mRNA) oncogenic transcripts. As the Aptima assay identifies oncogenic expression, it should have a lower rate of hrHPV and genotype detection. METHODS: The Kaiser Permanente Regional Reference Laboratory in Denver, Colorado changed its hrHPV assay from the cobas to the Aptima assay. The rates of hrHPV detection and genotyping were compared over successive six-month periods. RESULTS: The overall hrHPV detection rates by the two platforms were similar (9.5% versus 9.1%) and not statistically different. For genotyping, the HPV 16 rate by the cobas was 1.6% and by the Aptima it was 1.1%. These differences were statistically different with the Aptima detecting nearly one-third less HPV 16 infections. With the HPV 18 and HPV 18/45, there was a slightly higher detection rate of HPV 18/45 by the Aptima platform (0.5% versus 0.9%) and this was statistically significant. CONCLUSION: While HPV 16 represents a low percentage of hrHPV infections, it was detected significantly less by the Aptima assay compared to the cobas assay. This has been previously reported, although not highlighted. Given the test methodologies, one would expect the Aptima to detect less HPV 16. This difference appears to be mainly due to a significantly increased number of non-oncogenic HPV 16 infections detected by the cobas test as there were no differences in HPV 16 detection rates in the high-grade squamous intraepithelial lesions indicating that the two tests have similar sensitivities for oncogenic HPV 16.

The use of the papanicolaou stain to routine histological examination: teaching the basic microscopy of squamous cell carcinoma

Introduction: The understanding of microscopy classes is imperative to the student in human pathology. Objective: To prepare the student to observe and understand, through tinctorial recognition, the main histologic features of Papanicolaou-stained well-differentiated carcinomas. Methods: A selected case of a well-differentiated carcinoma of the tongue was used for a Papanicolaou stain. Results: The Papanicolaou-stained slides showed, basically, basal cells in a faint to a dark blue color. The stratum spinosum stained light pink to a faint blue-green and the keratin layer and the keratin pearls stained bright orange. The connective tissue stained light green. Conclusion: The use of Papanicolaou stain was found very convenient as the beginner in histopathology can perform a ready identification of the well-differentiated squamous cell carcinoma aspects.

Comparison of glass slides and various digital-slide modalities for cytopathology screening and interpretation.

BACKGROUND: Whole-slide imaging in cytology is limited when glass slides are digitized without z-stacks for focusing. Different vendors have started to provide z-stacking solutions to overcome this limitation. The Panoptiq imaging system allows users to create digital files combining low-magnification panoramic images with regions of interest (ROIs) that are imaged with high-magnification z-stacks. The aim of this study was to compare such panoramic images with conventional whole-slide images and glass slides for the tasks of screening and interpretation in cytopathology. METHODS: Thirty glass slides, including 10 ThinPrep Papanicolaou tests and 20 nongynecologic cytology cases, were digitized with an Olympus BX45 integrated microscope with an attached Prosilica GT camera. ViewsIQ software was used for image acquisition and viewing. These glass slides were also scanned on an Aperio ScanScope XT at ×40 (0.25 µm/pixel) with 1 z-plane and were viewed with ImageScope software. Digital and glass sides were screened and dotted/annotated by a cytotechnologist and were subsequently reviewed by 3 cytopathologists. For panoramic images, the cytotechnologist manually created digital maps and selected representative ROIs to generate z-stacks at a higher magnification. After 3-week washout periods, panoramic images were compared with Aperio digital slides and glass slides. RESULTS: The Panoptiq system permitted fine focusing of thick smears and cell clusters. In comparison with glass slides, the average screening times were 5.5 and 1.8 times longer with Panoptiq and Aperio images, respectively, but this improved with user experience. There was no statistical difference in diagnostic concordance between all 3 modalities. Users' diagnostic confidence was also similar for all modalities. CONCLUSIONS: The Aperio whole-slide scanner with 1 z-plane scanning and the Panoptiq imaging system with z-stacking are both suitable for cytopathology screening and interpretation. However, ROI z-stacks do offer a superior mechanism for overcoming focusing problems commonly encountered with digital cytology slides. Unlike whole-slide imaging, the acquisition of representative z-stack images with the Panoptiq system requires a trained cytologist to create digital files. Cancer Cytopathol 2017;125:701-9. © 2017 American Cancer Society.

Implementation of a real-time reference and calibration grid platform for improved screening - mapping in Pap test slides.

Cervical cancer screening based on the Papanicolaou (Pap) test is a widely applied but not always efficient practice for detecting Human Papillomavirus (HPV) mediated lesions, partially due to a non-systematic and inadequate screening process. Our aim was to introduce an inexpensive easy-to-use direct screening platform for improved detection of abnormal cells indicative of underlying cervical neoplasia as well as persisting HPV infection. By employing a novel, efficient technique of laser-based micromachining, we achieved the fabrication of spatial grids on commercially available coverslips allowing visual segmentation of the slide for efficient screening. Abnormal and formerly diagnosed as negative for intraepithelial lesion or malignancy (NILM) Pap test slides (n = 200) were analyzed by conventional and grid-based screening. Grid-based microscopy led to a more reliable diagnosis compared to the conventional by identifying an increased number of abnormal cells (P = 0.001). It decreased borderline ASCUS, AGC diagnosis, increasing LSIL, HSIL and in situ AdenoCa detection rates closely related with biopsy (P = 0.015; kappa = 0.978). Concerning the set of NILM diagnoses in rapid re-screening, the method upgraded six cases (n = 6) to LSIL (P = 0.001). The proposed technical solution offers a calibration and orientation visual aid during the on-site screening process providing significant advantages compared to expensive digital imaging techniques.

Indicador preventivo de saúde da mulher: proposta combinada de mamografia e Papanicolaou / Women's health prevention indicator: a proposal for combining mammography and Papanicolaou smear / Indicador preventivo de salud de la mujer: propuesta combinada de mamografía y prueba de Papanicolaou

RESUMO O presente artigo propõe um Indicador Preventivo de Saúde da Mulher (IPSM) que reflete o status combinado de realização de mamografia e Papanicolaou conforme as recomendações para a idade e considerando o tempo decorrido desde o último exame. A saúde preventiva foi categorizada em desejável, alerta e risco. Estão em risco as mulheres de qualquer idade que nunca fizeram o Papanicolaou, aquelas com >60 anos que o fizeram há mais de 3 anos mas nunca fizeram mamografia e aquelas com ≥71 anos que estão em dia com o Papanicolaou mas nunca fizeram mamografia. A condição desejável inclui mulheres com Papanicolaou de menos de 3 anos, exceto mulheres com ≥41 anos que nunca fizeram mamografia e com ≥51 anos que realizaram mamografia há mais de 2 anos. A condição de alerta inclui mulheres com Papanicolaou há mais de 3 anos, exceto aquelas com ≥61 que nunca fizeram mamografia e as com ≥71 anos com mamografia de mais de 2 anos. Para Papanicolaou há menos de 3 anos, a categoria alerta inclui mulheres entre 41 e 50 anos que nunca fizeram mamografia, as de 51 a 70 anos que fizeram mamografia há mais de 2 anos ou nunca fizeram e as com ≥71 anos com mamografia de mais de 2 anos. Aplicando-se o IPSM aos dados da Pesquisa Nacional por Amostra de Domicílios de 2008, constatou-se que 24,8% das mulheres no Brasil estavam em condição de risco e 24,2%, de alerta. Nordeste e Norte apresentaram as maiores proporções de risco (31,5% e 29,6%, respectivamente). Entre mulheres acima de 70 anos, 49,5% estavam em risco. O IPSM pode ser utilizado para avaliar ações públicas e para comparar o padrão preventivo dentro e entre regiões.(AU)

ABSTRACT This article proposes a women's health prevention indicator (WHPI) reflecting the combined status of mammography and Papanicolaou (Pap) smear according to the recommendations for age and considering the time elapsed since the last exam/test. The WHPI classifies prevention status into desirable, alert, or risk categories. The risk category includes women of all ages who never had a Pap smear, those aged >60 years who had a Pap smear more than 3 years ago but never had a mammography, and those aged ≥71 years who are up to date with the Pap smear but never had a mammography. The desirable category includes women with a Pap smear in the past 3 years, except women aged ≥41 who never had a mammography and those aged ≥51 years who had a mammography more than 2 years earlier. The alert category includes women whose Pap smear is more than three years old with the exception of those ≥61 years who never had a mammography and those aged ≥71 years whose mammography is more than 2 years old. For women who had experienced a Pap smear in the past 3 years, the alert category includes those aged 41-50 years who never had mammography, those aged 51-70 years with mammography older than 2 years or no mammography, and those aged ≥71 years with mammography older than 2 years. Applying the WHPI to data from the National Household Sample Survey of 2008 revealed that 24.8% of Brazilian women were at risk and 24.2% were in the alert category. The Northeast and the North had the highest risk rates (31.5% and 29.6% respectively). Of those >70 years old, 49.5% were in the risk category. The WHPI can be used to assess public initiatives and compare preventive status within and across regions.(AU)

RESUMEN En este artículo se propone un indicador preventivo de salud de la mujer (IPSM) que indique de manera combinada la situación respecto a la realización de una mamografía y de una prueba de Papanicolaou, según las recomendaciones para la edad y considerando el tiempo transcurrido desde el último examen. La situación de salud preventiva se clasificó en deseable, en alerta y en riesgo. Están en riesgo las mujeres de cualquier edad que nunca se han realizado la prueba de Papanicolaou, las mayores de 60 años que se la hicieron hace más de tres años, pero que nunca se han hecho una mamografía, y las de 71 años o más que están al día con el Papanicolaou, pero nunca se han hecho una mamografía. Están en una situación deseable las mujeres que se han hecho el Papanicolaou hace menos de 3 años, excepto las de 41 años o más que nunca se han hecho una mamografía y las de 51 años o más que hace más de dos años que se han hecho una mamografía. La situación de alerta se aplica a las mujeres que se han hecho el Papanicolaou hace más de 3 años, excepto las de 61 años o más que nunca se han sometido a una mamografía y las de 71 años o más que hace más de dos años que se hicieron una mamografía. Entre las que se han hecho el Papanicolaou hace menos de tres años, están en situación de alerta las mujeres de 41 a 50 años que nunca se han hecho una mamografía, las de 51 a 70 años que se han hecho una mamografía hace más de dos años y las de 71 años o más que hace más de dos años que se hicieron una mamografía. Al aplicar el IPSM a los datos de la encuesta nacional por muestra de hogares del 2008, se constató que un 24,8% de las mujeres brasileñas estaban en situación de riesgo y un 24,2% en situación de alerta. Las zonas nordeste y norte presentaban las mayores proporciones de riesgo (31,5% y 29,6%, respectivamente). En las mujeres mayores de 70 años, el 49,5% estaba en situación de riesgo. El IPSM puede utilizarse para evaluar acciones públicas y para comparar el patrón de prevención dentro de cada zona y entre las distintas zonas.(AU)

Application of Returned Cell Block Method (Cell Block from a Papanicolaou Staining Smear on a Glass Slide) for the Evaluation of Fine Needle Aspiration Cytology of Tumors of the Breast.

BACKGROUND: In the fine needle aspiration cytology (FNAC) for tumors of the breast, evaluation is frequently difficult because of the thick-layered cell clusters and blood inclusion. Such problems may be resolved by the returned cell block method, but its use has not spread worldwide. Here, we examined the application of the returned cell block method to cases involving difficulty in the evaluation of FNAC to diagnose tumors of the breast. METHODS: In Juntendo University Nerima Hospital, there were 22 cases which were difficult to diagnose by Papanicolaou stain only, and they underwent additional examination using the returned cell block method (cell block from a Papanicolaou staining smear on a glass slide). The usefulness of the returned cell block method in these cases was examined. RESULTS: Among the 22 cases, a correct diagnosis was facilitated in 20 cases using the returned cell block method. In 16 of the 20 cases, the difficulty in FNAC was because of thick-layered cell clusters (12 cases) and blood inclusion (four cases). Among the 12 cases with difficulty because of the thick-layered cell clusters, 10 cases (83%) comprised intraductal papilloma (six cases) and intraductal papillary carcinoma (four cases). Papilloma and papillary carcinoma were correctly diagnosed by the addition of histological images and immunostaining of myoepithelial cells using the returned cell block method. CONCLUSION: The application of the returned cell block method is useful for precise evaluation of the cytological diagnosis of tumors of the breast, especially papillary lesions. Diagn. Cytopathol. 2016;44:505-511. © 2016 The Authors Diagnostic Cytopathology Published by Wiley Periodicals, Inc.

Maximizing the adequacy of Hologic(®) Cervista(®) HPV HR results on ThinPrep(®) Pap samples treated with glacial acetic acid.

BACKGROUND: Although glacial acetic acid (GAA) treatment of bloody cervical samples has reduced the rate of unsatisfactory Pap Tests, recent studies suggest that it may negatively impact high-risk (hr)-HPV test results. The objectives of this study were to compare the levels of genomic DNA between GAA treated and nontreated ThinPrep(®) samples using the Hologic(®) Cervista(®) HPV-HR assay and to compare the adequacy of the ThinPrep(®) Pap Test between aliquoted and nonaliquoted samples. METHODS: Prior to GAA treatment, 2.5 ml of the cervical sample was prealiquoted from 102 bloody ThinPrep(®) vials. Both GAA treated and nontreated samples were analyzed for hr-HPV using the Cervista(®) HPV HR assay. The levels of genomic DNA were measured and compared between these samples. In addition, ThinPrep(®) Pap Test adequacy rates were calculated and compared on aliquoted and nonaliquoted cervical samples. RESULTS: Of the 102 cervical samples, 95 (93%) nontreated aliquots contained satisfactory levels of genomic DNA as compared to 36 (35%) GAA-treated samples (p < 0.00001). Ninety-nine (97%) aliquoted cervical samples were satisfactory for cytologic evaluation as compared to 1,326 (96%) GAA treated samples from 2013 (nonaliquoted); not statistically significant (p = 0.7968). CONCLUSION: The levels of genomic DNA were significantly decreased in GAA treated vs non-treated TP samples. Aliquoting from the TP sample prior to treatment with GAA enables accurate measurement of DNA without affecting the adequacy of the TP cytology slide.

Cost-effectiveness of using the Cervex-Brush (broom) compared to the elongated spatula for collection of conventional cervical cytology samples within a high-burden HIV setting: a model-based analysis.

BACKGROUND: From 2010 to 2014, approximately 2 million Pap smears from HIV-infected women were submitted to the South African National Health Laboratory Services (NHLS) through the national cervical cancer screening programme. The objective of this analysis was to determine whether using the plastic Cervex brush ("broom") would be a cost-effective approach to improve cytology specimen quality as compared to the wooden spatula used currently. METHODS: A decision analysis model was built using the expected adequacy rates for samples collected with the spatula (<$0.02) and broom ($0.23) and the probability of detecting cervical dysplasia. NHLS data was used for testing volumes and rates of HIV-positivity, suitability of specimens, and presence of endocervical cells. Expected positivity of Pap smears in HIV-infected women (73 %), odds ratios of the effectiveness of the broom (OR: 1.57), and improved sensitivity when endocervical cells present (OR: 1.89) are from literature. NHLS costs were used for the collection devices and conventional cytology ($4.89). Cost of clinic visit is from WHO CHOICE ($8.36). RESULTS: In 2010, 80 % of specimens submitted to NHLS were adequate for evaluation; in 2014, only 54 % met the same criteria. For HIV-infected women, according to the guidelines model, using the wooden spatula costs $6.25 million per year, $16.79 per woman tested. Under intended practice, for each additional HSIL case detected among HIV-infected women, the South African cervical cancer screening programme could save $13.64 (95 % CI: $13.52 to $13.76) by using the broom as its standard of care collection device through increased collection of endocervical cells and consequent reduction in repeat Pap smears. CONCLUSION: Under a wide range of parameters tested using a simulation model, the more expensive plastic broom could save the South African cervical cancer screening programme money and increase detection of high-grade cervical dysplasia in HIV-infected women compared to the current wooden spatula.

Proflavine Hemisulfate as a Fluorescent Contrast Agent for Point-of-Care Cytology.

Proflavine hemisulfate, an acridine-derived fluorescent dye, can be used as a rapid stain for cytologic examination of biological specimens. Proflavine fluorescently stains cell nuclei and cytoplasmic structures, owing to its small amphipathic structure and ability to intercalate DNA. In this manuscript, we demonstrated the use of proflavine as a rapid cytologic dye on a number of specimens, including normal exfoliated oral squamous cells, cultured human oral squamous carcinoma cells, and leukocytes derived from whole blood specimens using a custom-built, portable, LED-illuminated fluorescence microscope. No incubation time was needed after suspending cells in 0.01% (w/v) proflavine diluted in saline. Images of proflavine stained oral cells had clearly visible nuclei as well as granular cytoplasm, while stained leukocytes exhibited bright nuclei, and highlighted the multilobar nature of nuclei in neutrophils. We also demonstrated the utility of quantitative analysis of digital images of proflavine stained cells, which can be used to detect significant morphological differences between different cell types. Proflavine stained oral cells have well-defined nuclei and cell membranes which allowed for quantitative analysis of nuclear to cytoplasmic ratios, as well as image texture analysis to extract quantitative image features.

Simulation of bright-field microscopy images depicting pap-smear specimen.

As digital imaging is becoming a fundamental part of medical and biomedical research, the demand for computer-based evaluation using advanced image analysis is becoming an integral part of many research projects. A common problem when developing new image analysis algorithms is the need of large datasets with ground truth on which the algorithms can be tested and optimized. Generating such datasets is often tedious and introduces subjectivity and interindividual and intraindividual variations. An alternative to manually created ground-truth data is to generate synthetic images where the ground truth is known. The challenge then is to make the images sufficiently similar to the real ones to be useful in algorithm development. One of the first and most widely studied medical image analysis tasks is to automate screening for cervical cancer through Pap-smear analysis. As part of an effort to develop a new generation cervical cancer screening system, we have developed a framework for the creation of realistic synthetic bright-field microscopy images that can be used for algorithm development and benchmarking. The resulting framework has been assessed through a visual evaluation by experts with extensive experience of Pap-smear images. The results show that images produced using our described methods are realistic enough to be mistaken for real microscopy images. The developed simulation framework is very flexible and can be modified to mimic many other types of bright-field microscopy images.

Cervicovaginal cytology in patients undergoing pelvic radiotherapy using the Focalpoint system: results from the RODEO study.

BACKGROUND: Evaluate the performance of the Focalpoint system in identifying and classifying cervical cytology alterations from samples collected from patients treated with Radiotherapy (RT). METHODS: The reproducibility of manual and automated screening by cytotechnologists using the BD Focalpoint GS Imaging System was examined. Samples were collected from May 2010 to August 2011. RESULTS: A total of 378 treated with RT and 8,967 cytology samples from patients without previous RT, were evaluated. The kappa values for cytological diagnoses read manually and automated in cases without previous RT were as follows: < ASC-H vs. ≥ ASC-H = 0.71; < LSIL vs. ≥ LSIL = 0.66; and < HSIL vs. ≥ HSIL = 0.67. The kappa for cytological diagnoses in post-RT women have showed: < ASC-H vs. ≥ ASC-H = 0.71; < LSIL vs. ≥ LSIL = 0.65; < HSIL vs. ≥ HSIL = 0.57. CONCLUSIONS: There was no significant difference among the kappa values we found. Post-RT cytology showed small diagnostic agreement between manual and automated examination. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/13000_2014_231.

Caracterização imunocitoquímica da expressão da proteína RAP1 em blocos de células escamosas provenientes de citologia cervical em meio líquido

O diagnóstico precoce acurado do câncer cervical, relevante problema de saúde pública no mundo e no Brasil, pela citologia oncótica (Teste de Papanicolaou), é muito prejudicado pela subjetividade e variabilidade dos resultados falsos negativos e falsos positivos do método, particularmente diante de células escamosas atípicas(ASC). Recentes inovações técnicas, como citologia em meio líquido e imunocitoquímica com biomarcadores de proliferação celular, aumentaram a expectativa de melhorias no rastreamento do câncer cervical. No entanto, a aplicabilidade destas inovações nos estágios mais iniciais de atipia e displasias epiteliais permanecem incertas. Assim, considerando resultado prévio do nosso grupo de pesquisa, que identificou a proteína RAP1 como biomarcador diagnóstico da displasia cervical, este trabalho tem como objetivo caracterizar a expressão da proteína RAP1, comparativamente à expressão dos biomarcadores p16 e Ki-67, por imunocitoquímica, em blocos de células escamosas cervicais para possível aplicabilidade na triagem do câncer do colo do útero. Para tal, 34 amostras, 27 pacientes com diagnóstico de alterações celulares benignas (ACB) e 7 pacientes com diagnóstico de ASC foram coletadas na unidade Jenny Faria do Hospital das Clínicas da UFMG. Os resultados indicaram que 85% das amostras de blocos celulares foram satisfatórias para análise morfológica e a técnica reproduziu os principais parâmetros citopatológicos da citologia convencional.

Em relação a sua utilização para diagnóstico, o bloco de células apresentou sensibilidade de 38,46%,especificidade de 90,47% e uma variabilidade interobservador com taxa de concordância de aproximadamente 30% para os grupos ACB e ASC. A expressão da proteína RAP1 foi positiva na maioria das amostras do grupo “ACB” (15/27 ou55,56%) e resultado negativo na maioria das amostras do grupo “ASC” (4/7 ou 57,14%) com sensibilidade de 16,66%, especificidade de 75, %. As reações imunocitoquímicas das proteínas p16 e Ki-67 demonstraram, em ambos os grupos,apresentaram predomínio absoluto ou totalidade de resultados negativos. O DNA de HPV foi detectado em 9 (33,33%) das 27 amostras do grupo ACB e em 4 (57,14%)das 7 amostras do grupo ASC. O HPV-16 foi detectado nas 4 amostras do grupoASC. Nas amostras do grupo ACB foram detectados os HPV-16, em 5 amostras,HPV-58, em 2 amostras, HPV-45, em 1 amostra e HPV-66 em 1 amostra.Observamos inexistência de relação entre a presença do HPV e a expressão imunocitoquímica de RAP1 em ambos os grupos. Em conclusão, os blocos de células podem complementar o teste de Papanicolaou na triagem do câncer do colo uterino e a expressão da proteína RAP1 está aumentada em células cervicais em ambiente inflamatório, associado ou não à presença de HPV.

Caracterização imunocitoquímica da expressão da proteína RAP1 em blocos de células escamosas provenientes de citologia cervical em meio líquido / Immunocytochemistry characterization of expression of RAP1 protein in squamous cell blocks from cervical cytology in liquid medium

O diagnóstico precoce acurado do câncer cervical, relevante problema de saúde pública no mundo e no Brasil, pela citologia oncótica (Teste de Papanicolaou), é muito prejudicado pela subjetividade e variabilidade dos resultados falsos negativos e falsos positivos do método, particularmente diante de células escamosas atípicas(ASC). Recentes inovações técnicas, como citologia em meio líquido e imunocitoquímica com biomarcadores de proliferação celular, aumentaram a expectativa de melhorias no rastreamento do câncer cervical. No entanto, a aplicabilidade destas inovações nos estágios mais iniciais de atipia e displasias epiteliais permanecem incertas. Assim, considerando resultado prévio do nosso grupo de pesquisa, que identificou a proteína RAP1 como biomarcador diagnóstico da displasia cervical, este trabalho tem como objetivo caracterizar a expressão da proteína RAP1, comparativamente à expressão dos biomarcadores p16 e Ki-67, por imunocitoquímica, em blocos de células escamosas cervicais para possível aplicabilidade na triagem do câncer do colo do útero. Para tal, 34 amostras, 27 pacientes com diagnóstico de alterações celulares benignas (ACB) e 7 pacientes com diagnóstico de ASC foram coletadas na unidade Jenny Faria do Hospital das Clínicas da UFMG. Os resultados indicaram que 85% das amostras de blocos celulares foram satisfatórias para análise morfológica e a técnica reproduziu os principais parâmetros citopatológicos da citologia convencional...

Em relação a sua utilização para diagnóstico, o bloco de células apresentou sensibilidade de 38,46%,especificidade de 90,47% e uma variabilidade interobservador com taxa de concordância de aproximadamente 30% para os grupos ACB e ASC. A expressão da proteína RAP1 foi positiva na maioria das amostras do grupo “ACB” (15/27 ou55,56%) e resultado negativo na maioria das amostras do grupo “ASC” (4/7 ou 57,14%) com sensibilidade de 16,66%, especificidade de 75, %. As reações imunocitoquímicas das proteínas p16 e Ki-67 demonstraram, em ambos os grupos,apresentaram predomínio absoluto ou totalidade de resultados negativos. O DNA de HPV foi detectado em 9 (33,33%) das 27 amostras do grupo ACB e em 4 (57,14%)das 7 amostras do grupo ASC. O HPV-16 foi detectado nas 4 amostras do grupoASC. Nas amostras do grupo ACB foram detectados os HPV-16, em 5 amostras,HPV-58, em 2 amostras, HPV-45, em 1 amostra e HPV-66 em 1 amostra.Observamos inexistência de relação entre a presença do HPV e a expressão imunocitoquímica de RAP1 em ambos os grupos. Em conclusão, os blocos de células podem complementar o teste de Papanicolaou na triagem do câncer do colo uterino e a expressão da proteína RAP1 está aumentada em células cervicais em ambiente inflamatório, associado ou não à presença de HPV...

Evaluation of CellSolutions BestPrep® automated thin-layer liquid-based cytology Papanicolaou slide preparation and BestCyte® cell sorter imaging system.

OBJECTIVE: A double-blind study was conducted to compare the performance of the new BestPrep® (CellSolutions) liquid-based thin-layer Papanicolaou (Pap) test with ThinPrep® (Hologic). STUDY DESIGN: Samples from the study patients (n = 105) were collected twice in the same encounter with the ThinPrep sample always taken first and the BestPrep sample collected second. Slides were prepared according to both manufacturers' protocols and evaluated using manual microscopic review and the BestCyte® cell sorter imaging system (CellSolutions). Diagnostic truth for each case was determined by independent manual review of both slides by multiple pathologists and histology when available. The presence of atypical squamous cells of undetermined significance was the threshold for positive for sensitivity and specificity calculations. RESULTS: BestPrep and ThinPrep, by manual review, had sensitivities for high-grade squamous intraepithelial lesion (HSIL) cases of 100 and 95.6%, respectively. Using the BestCyte cell sorter, both had 100% sensitivity. For the same HSIL cases, the digene HC2 high-risk human papillomavirus DNA test had sensitivities of 100% (BestPrep) and 95.6% (ThinPrep). Specificities were 71.4% (BestPrep) and 54.8% (ThinPrep). CONCLUSIONS: BestPrep was equivalent to ThinPrep for manual review even though BestPrep was always the second sample collected. The BestCyte cell sorter provides a practical alternative to manual review for both BestPrep and ThinPrep slides.