Prevalencia y genotipicación de virus papiloma humano vaginal y cervical en trabajadoras sexuales de un centro de salud sexual en la zona Norte de Santiago, Chile / Prevalence and genotypes of cervical and vaginal HPV in female sex workers attending a Sexual Control Centre, North Area of Santiago, Chile

INTRODUCCIÓN: En Chile, el cáncer de cuello uterino (CCU) es la segunda causa de muerte por neoplasias malignas en la mujer. El principal agente causal es el virus papiloma humano (VPH). Comparando con la población general, los o las trabajadoras(es) sexuales (TS) tienen alto riesgo de adquirir VPH. OBJETIVO: Analizar la prevalencia y genotipos del VPH cervical y vaginal en TS que se atienden en un Centro de Salud Sexual de Santiago, Chile. Pacientes y MÉTODO: Se realizó un estudio transversal en 97 mujeres TS, de 19 a 70 años de edad. Se obtuvieron dos muestras por paciente, una de exocérvix y otra de paredes vaginales. El ADN de VPH fue identificado por reacción de polimerasa en cadena (RPC) y su genotipo fue investigado para 32 tipos de VPH. RESULTADOS: La prevalencia de VPH global fue de 45%, observándose portación cervical en 41,2% y vaginal en 36,1%, con una coinfección de 32%. El 63% de las muestras tenía genotipos de alto riesgo. Los VPH de alto riesgo más frecuentes fueron el VPH 66 (12%), VPH 58 (9,3%), seguidos por VPH 16, VPH 59 y VPH 82 con igual frecuencia (8% c/u). Treinta y dos mujeres (43%) fueron infectadas con genotipos múltiples. CONCLUSIÓN: El VPH es una infección frecuente entre las TS. Este es el primer estudio en Chile sobre prevalencia y genotipos de VPH en TS.

BACKGROUND: In Chile, cervical cancer is the second leading cause of death from malignancy in women. The main causal agent of cervical cancer is the human papillomavirus (HPV). Compared with the general population, sex workers (SW) are at increased risk of acquiring HPV. AIM: To analyze the prevalence and genotypes of cervical and vaginal HPV in female SW attending a Sexual Control Centre. METHODS: A cross-sectional study was carried out on 97 women (19-70 years old). Two samples were taken per patient, one from exocervix and the other from vaginal walls. HPV DNA. was identified by polymerase chain reaction (PCR) and genotyping using specific probes for 32 types of HPV. RESULTS: The overall frequency of HPV was 45%, 41.2% in cervical carrier and 36.1% in vaginal carrier, 32% were co-infected, 63% of HPV were high-risk genotypes. The most frequent high-risk HPV was HPV 66 (12%), HPV 58 (9.3%), followed by HPV 16, HPV 59 and HPV 82 with the same frequency (8% each one). Thirty two (43%) of females were infected with multiple genotypes. CONCLUSION: HPV is frequent infection among SW. This is the first study in Chile on the prevalence and genotypes of HPV in sex workers.

Influencia del cáncer cérvico-uterino y la alteración de la salud mental en pacientes menores de 25 años en Ecuador / Influence of cervico-uterine cancer and the alteration of mental health in patients under 25 years of age in Ecuador

INTRODUCCIÓN: Los problemas sanitarios en los pacientes que padecen cáncer cérvico-uterino influyen en la satisfacción personal y evitan una gran adherencia a la terapia médica, y además están relacionados con un número más notable de efectos secundarios y la estancia en la clínica de emergencia junto afecciones depresivas y problemas de ansiedad. OBJETIVO: Identificar la influencia del cáncer cérvico -uterino en las alteraciones en la salud mental de pacientes menores de 25 años en el Ecuador. METODOLOGÍA: La investigación fue de tipo cuantitativo, descriptivo, y de corte transversal. Se utilizaron encuestas de manera virtual dirigida a pacientes menores de 25 años que padecen de cáncer al cuello uterino en diversas Fundaciones que luchan contra el cáncer, contando con una muestra de 178 mujeres, estas se dividieron en 4 categorías: Salud mental, Estrés, Depresión y Total de escalas. RESULTADOS: El porcentaje que se obtuvo de manera general tuvo un rango variado, el 32,9% dio como resultado que la alteración que causa en la mujer es contundente, seguido del 30,6 que su afección en la salud mental es más baja. CONCLUSIÓN: Se concluye afirmando que el cáncer uterino es el segundo cáncer más prevalente en la mujer, el cual influye de manera progresiva en la salud mental, presentando reacciones negativas y percepciones, falta de control en las emociones, consecuentemente aquello debilita el sistema inmunológico previamente comprometido, generando alteraciones en el curso del tratamiento.

INTRODUCTION: Health problems in patients suffering from cervical-uterine cancer influence personal satisfaction and prevent a high adherence to medical therapy, and are also related to a more notable number of side effects and stay in the emergency clinic together depressive conditions and anxiety problems. OBJECTIVE: To identify the influence of cervical cancer -uterine in the alterations in the mental health of patients under 25 years in Ecuador. MATERIALS AND METHODS: The research was quantitative, descriptive, andcross-sectional. Surveys were used in a virtual way directed at patients under 25 years of age who suffer from cervical cancer in various Foundations that fight against cancer, with a sample of 178 women, these were divided into 4 categories: Mental health, Stress, Depression and Total stopovers. RESULTS: The percentage that was obtained in a general way had a varied range, 32.9% gave as a result that the alteration it causes in women is overwhelming, followed by 30.6 that their mental health condition is lower. CONCLUSION: It is concluded by stating that uterine cancer is the second most prevalent cancer in women, which progressively influences mental health, presenting negative reactions and perceptions, lack of control over emotions, consequently that weakens the previously compromised immune system, generating alterations in the course of treatment.

Prevalencia de los genotipos de HPV en lesiones pre invasoras de alto grado de malignidad y cáncer de cuello uterino en la población del Hospital de Clínicas: Montevideo-Uruguay / Prevalence of HPV genotypes in cervical high risk intraepithelial lesions and cervical cancer in the population of Hospital de Clínicas: Montevideo-Uruguay / Prevalência de genótipos de HPV em lesões pré-invasivas com alto grau de malignidade e câncer de colo do útero na população do Hospital de Clínicas: Montevidéu, Uruguai

El Cáncer de Cuello Uterino (CCU) es un problema de Salud pública a nivel mundial. Su indiscutible asociación con el Virus del papiloma humano (HPV) hace necesario su estudio. El objetivo de este trabajo, es conocer la prevalencia de los diferentes genotipos de HPV, en lesiones pre invasoras de alto grado de malignidad (HSIL) y/o cáncer de cuello uterino. Material y Métodos: Todas las Mujeres que fueron derivadas a pol de TGI del H Clínicas entre enero del 2011 y diciembre de 2012, por un PAP sospechoso de lesión y en las que se confirmó luego un HSIL o cáncer cervical fueron tipificadas. Se recabaron datos de edad, tipo de lesión y genotipificación. La extracción de ADN viral se realizó a partir de muestras cervico vaginales conservadas en medio de transporte comercial (Sacace) mediante el kit QIAamp DNA Mini Kit (QIAGEN). Se buscaron 14 genotipos de alto riesgo. Resultados: Se tipificaron 75 pacientes, 19 con CCU y 56 con HSIL. El HPV 16 fue el más prevalente en un 61,5 % para las infecciones únicas y en un casi 100 % para las múltiples y un 60% para el total de las lesiones, seguido en prevalencia por los HPV 31,33 y 45. El HPV 18 fue muy poco prevalente. Conclusiones: En esta muestra, la prevalencia del HPV 16 está acorde con las publicaciones nacionales siendo el más frecuente. El HPV 18 tiene muy baja prevalencia siendo 2 casos en 75, siempre en infecciones múltiples.

Cervical Cancer (CC) is a public health problem worldwide. Its indisputable association with the Human Papilloma Virus (HPV) makes its study necessary. The objective of this work is to know the prevalence of the different HPV genotypes, in pre-invasive high-grade malignant lesions (HSIL) and / or cervical cancer. Material and Methods: All women who were referred to Low genital tract service in the Hospital de Clinicas between January 2011 and December 2012, for a PAP suspected of injury and in which HSIL or cervical cancer was later confirmed were typified. Data on age, type of lesion and genotyping were collected. The viral DNA extraction was carried out from cervico-vaginal samples preserved in commercial transport medium (Sacace) using the QIAamp DNA Mini Kit (QIAGEN). 14 high-risk genotypes were searched. Results: 75 patients were typified, 19 with CCU and 56 with HSIL. HPV 16 was the most prevalent in 61.5% for single infections and almost 100% for multiple infections and 60% for all lesions, followed in prevalence by HPV 31,33 and 45. The HPV 18 was very rare. Conclusions: In this sample, the prevalence of HPV 16 is in accordance with national publications, being the most frequent. HPV 18 has a very low prevalence, being 2 cases in 75, always in multiple infections.

O câncer cervical é um problema de saúde pública em todo o mundo. E um câncer com uma ligação comprovada com o vírus do papiloma humano. O objetivo é conhecer a prevalência dos diferentes genótipos do HPV em mulheres que apresentam neoplasias pré-invasivas de alto grau e câncer de colo do útero, que foram tratadas e diagnosticadas no Hospital de Clínicas entre janeiro de 2011 e dezembro de 2012. Material e Métodos: o estudo foi realizado em 75 pacientes do Hospital de Clinicas, com diagnóstico histológico de Câncer Cervical e lesões intraepiteliais de alto grau para as quais foi realizado o tipageme do HPV alto risco. Resultados: 75 pacientes foram tipificados, 19 com CCU e 56 com HSIL. O HPV 16 foi o mais prevalente em 61,5% para infecções únicas e quase 100% para infecções múltiplas e 60% para todas as lesões, seguido em prevalência pelo HPV 31,33 e 45. O HPV 18 era muito raro. Conclusões: Nesta amostra, a prevalência do HPV 16 está de acordo com as publicações nacionais, sendo a mais frequente. O HPV 18 tem prevalência muito baixa, sendo 2 casos em 75, sempre em infecções múltiplas.

Medidas de autocuidado y genotipificación del virus papiloma humano en mujeres de la Unidad de Patología Cervical, Hospital Carlos Van Buren / Self-care measures and genotyping of human papillomavirus in women from Cervical Pathology Unit, Hospital Carlos Van Buren

OBJETIVO: Identificar las características biosociales, medidas de autocuidado y genotipoficación del virus papiloma humano en mujeres con papanicolaou alterado en la primera consulta de la Unidad Patología Cervical, Hospital Carlos Van Buren, Valparaíso, 2013. MATERIAL Y MÉTODO: Cuantitativo, descriptivo y transversal. Aprobada por Comité Ético-Científico. Muestra de 50 mujeres que previa firma de consentimiento informado, se les aplicó un cuestionario y se les tomó muestras cervicales para genotipificación VPH con técnica PCR e Hibridación. RESULTADOS: 26% con edad entre 25-34 años, 58% tiene a lo menos cuarto medio, 66% tuvieron primera relación sexual entre los 15-19 años, 48% presentaron test positivos para VPH alto riesgo, 26% tiene antecedentes de otras infecciones de transmisión sexual, 26% no se realiza el Pap de forma regular y 90% no utiliza preservativo. DISCUSIÓN: Conocer algunas características biosociales de la población permite examinar estrategias gubernamentales en la prevención de la adquisición del VPH y por ende del desarrollo de cáncer cervicouterino, como son: educación sexual, estilos de vida saludables, estrategias para adhesión a la toma del Pap, entre otras. Además se hace necesario ampliar estudios e investigaciones en estos temas que permitan contribuir en la vida sexual de la mujer y de la sociedad.

OBJECTIVE: To identify the biosocial characteristics, self-care measures and human papillomavirus (HPV) genotypes in women with altered Pap at the first consultation at the Cervical Pathology Unit, in Hospital Carlos Van Buren, Valparaiso, 2013. METHODS: Quantitative, descriptive and transversal. Approved by the Ethics Committee of Hospital Carlos Van Buren. Fifty women provided informed consent and answered a questionnaire. Cervical samples were taken for HPV genotyping performed by PCR and hybridization. RESULTS: 26% of women were aged between 25-34 years, 58% finished high school, 66% had their first sexual encounter between 15-19 years, 48% tested positive for high risk HPV, 26% have a history of other sexually transmitted infections, 26% do not undertake regular Pap exams and 90% do not use condoms. DISCUSSION: Knowledge of biosocial characteristics of this population permits examination of government strategies in HPV prevention and thus the development of cervical cancer, such as: sex education, healthy lifestyles, strategies for access to Pap testing, among others. In addition, it is necessary to expand research on issues that contribute to the sexual life of women and society.

DNA methylation in human papillomavirus-infected cervical cells is elevated in high-grade squamous intraepithelial lesions and cancer / 부인종양

OBJECTIVE: DNA methylation has been shown to be a potential biomarker for early cancer detection. The aim of this study was to evaluate DNA methylation profiles according to liquid-based Pap (LBP) test results and to assess their diagnostic value in a Korean population. METHODS: A total of 205 patients with various Papanicolaou test results were enrolled to this study (negative, 26; atypical squamous cells of undetermined significance, 39; low grade squamous intraepithelial lesion, 44; high grade squamous intraepithelial lesion (HSIL), 48; and cancer, 48). DNA methylation analysis of four genes, ADCYAP1, PAX1, MAL, and CADM1, was performed on residual cervical cells from LBP samples using a quantitative bisulfite pyrosequencing method. To evaluate the diagnostic performance of the four methylated genes for cancer detection, receiver operating characteristic (ROC) curves were drawn. Sensitivities and specificities were also tested at cutoffs determined from the ROC curves. RESULTS: Cervical cancer cells showed dramatically increased methylation levels for the four genes analyzed. ADCYAP1 and PAX1 also trended toward elevated methylation levels in HSIL samples, although the levels were much lower than those in cancer cells. The sensitivities of methylated ADCYAP1, PAX1, MAL, and CADM1 for the detection of cancer were 79.2%, 75.0%, 70.8%, and 52.1%, and the specificities were 92.0%, 94.0%, 94.7%, and 94.0%, respectively. Methylated ADCYAP1 and PAX1 demonstrated relatively better discriminatory ability than did methylated MAL and CADM1 (area under the curves 0.911 and 0.916 vs. 0.854 and 0.756, respectively). CONCLUSION: DNA methylation status, especially in the ADCYAP1 and PAX1 genes, showed relatively good specificity, ranging from 90% to 94%. The possible additive and complementary roles of DNA methylation testing with respect to conventional cervical cancer screening programs will need to be validated in prospective population-based studies.

Detección de virus papiloma humano en la prevención del cáncer cérvico-uterino / Human papilomavirus detection in cervical cancer prevention

El cáncer cérvico-uterino (CCU), que está fuertemente asociado a la infección por virus papiloma humano de alto riesgo (VPH-AR), sigue siendo un problema de salud pública en Latinoamérica. El uso de la citología para la detección de lesiones pre-cancerosas no ha tenido mayor impacto en las tasas de incidencia y mortalidad del CCU, que aún se mantienen altas en la región. La disponibilidad de nuevas técnicas de tamizaje para la detección de lesiones pre-cancerosas y de vacunas altamente eficaces que previenen casi todas las lesiones relacionadas con los VPH-AR de alto potencial oncogénico VPH 16 y 18, en mujeres no expuestas previamente al virus brindan una gran oportunidad para la prevención del CCU. La detección de VPH-AR representa actualmente un valioso componente de las guías clínicas para el tamizaje, manejo y tratamiento del CCU y sus lesiones precursoras. Se han desarrollado estrategias metodológicas que detectan un amplio espectro de tipos de VPH-AR; sin embargo, solo un pequeño subgrupo de ellas ha documentado la validación clínica para cualquiera de las indicaciones habituales de la detección de estos virus. Las pruebas de VPH que no estén validadas y que no hayan demostrado confiabilidad, reproducibilidad y exactitud no deben ser usadas en el manejo clínico. Una vez incorporada una prueba de VPH en el laboratorio, es esencial que el procedimiento completo sea sometido a un continuo y riguroso control de calidad para evitar prácticas subóptimas, potencialmente dañinas. Este artículo discute los recientes progresos y el estado actual de estos métodos.

Cervical cancer (CC), which is strongly associated to high-risk human papillomavirus (hr-HPV) infection, continues being a significant health problem in Latin America. The use of conventional cytology to detect precancerous cervical lesions has had no major impact on reducing CC incidence and mortality rates, which are still high in the region. New screening tools to detect precancerous lesions became available, which provide great opportunities for CC prevention, as do highly efficacious HPV vaccines able to prevent nearly all lesions associated with HPV-16 and -18 when applied before viral exposure. Currently, hr-HPV testing represents an invaluable component of clinical guidelines for screening, management and treatment of CC and their precursor lesions. Many testing strategies have been developed that can detect a broad spectrum of hr-HPV types in a single assay; however, only a small subset of them has documented clinical performance for any of the standard HPV testing indications. HPV tests that have not been validated and lack proof of reliability, reproducibility and accuracy should not be used in clinical management. Once incorporated into the lab, it is essential to submit the whole procedure of HPV testing to continuous and rigorous quality assurance to avoid sub-optimal, potentially harmful practices. Recent progress and current status of these methods are discussed in this article.

Human papillomavirus (HPV) detection and Papanicolaou cytology in low-resource women in Posadas city, Misiones, Argentina / Detección del virus papiloma humano (HPV) y citología de Papanicolaou en mujeres de bajos recursos de la ciudad de Posadas, Misiones, Argentina

The objective of this study was to determine the prevalence of HPV infection and cervical lesions present in women who attended a health center in a low-resource area of the city of Posadas, Misiones, Argentina. Cervical cell samples (n = 163) were processed for Papanicolaou cytology and HPV-PCR tests. Socio-cultural risk factors were estimated using the odds ratio (OR, CI 95 %). Cervical lesions were detected in 14.7 % of women. The general prevalence of HPV infection was of 38 %. The most common types among the total population were HPV-16 (9.8 %) and HPV-33 (9.3 %). HPV-16 was detected in association with 29.2 % and 6.5 % of women with and without cervical lesions, respectively, the OR being 5.3 (1.8-15.8). Risk factors for HPV-16 infection were a smoking habit and a history of previous sexually-transmitted diseases. These data are important for the implementation of prevention programs, including an appropriate introduction of vaccination and the baseline for virological surveillance in the vaccine era.

El objetivo de este estudio fue determinar la prevalencia de la infección por HPV y de lesiones cervicales en mujeres asistidas en un centro de salud situado en un área de bajos recursos de la ciudad de Posadas, Misiones, Argentina. Las muestras (n = 163) fueron examinadas mediante las pruebas de Papanicolaou y de PCR para HPV. Los factores socio-culturales de riesgo fueron identifcados mediante el cálculo de la odds ratio (OR, IC 95 %). Se detectaron lesiones cervicales en el 14,7 % de las mujeres. La prevalencia de infección por HPV fue de 38 %. Los tipos más frecuentes en la población total fueron HPV-16 (9,8 %) y HPV-33 (9,3 %). El HPV-16 se detectó asociado al 29,2 % y al 6,5 % de las mujeres con lesiones del cuello uterino y sin ellas, respectivamente, con un OR de 5,3 (1,8-15,8). Los factores de riesgo para la infección por HPV-16 fueron el hábito de fumar y el antecedente de enfermedades de transmisión sexual. Estos datos son importantes para la ejecución de los programas de prevención, incluyendo una introducción adecuada de la vacunación y la línea de base para la vigilancia virológica en la era de la vacuna.

Production of L1 protein from different types of HPV in Pichia pastoris using an integrative vector

Human papillomavirus (HPV) infection is the most common sexually transmitted disease in the world and is related to the etiology of cervical cancer. The most common high-risk HPV types are 16 and 18; however, the second most prevalent type in the Midwestern region of Brazil is HPV-33. New vaccine strategies against HPV have shown that virus-like particles (VLP) of the major capsid protein (L1) induce efficient production of antibodies, which confer protection against the same viral type. The methylotrophic yeast Pichia pastoris is an efficient and inexpensive expression system for the production of high levels of heterologous proteins stably using a wild-type gene in combination with an integrative vector. It was recently demonstrated that P. pastoris can produce the HPV-16 L1 protein by using an episomal vector associated with the optimized L1 gene. However, the use of an episomal vector is not appropriate for protein production on an industrial scale. In the present study, the vectors were integrated into the Pichia genome and the results were positive for L1 gene transcription and protein production, both intracellularly and in the extracellular environment. Despite the great potential for expression by the P. pastoris system, our results suggest a low yield of L1 recombinant protein, which, however, does not make this system unworkable. The achievement of stable clones containing the expression cassettes integrated in the genome may permit optimizations that could enable the establishment of a platform for the production of VLP-based vaccines.

Comparative analysis of three methods for HPV DNA detection in cervical samples / Análisis comparativo de tres métodos para la detección del ADN de VPH en muestras de cuello uterino

High risk HPV infection is considered to play a central role in cervical carcinogenesis. HPV DNA testing has shown to be a very useful tool for screening and following cervical infections. The aim of this study was to compare three methods for HPV DNA detection, along with cytology and colposcopy analysis. Cervical samples were collected from 100 sexually active women in Mérida, western Venezuela. HPV infection was screened using Hybrid-Capture 2 (HC2), L1-Nested-PCR and E6/E7-PCR assays. 40% of the samples (40/100) were HPV positive by at least one of the DNA detection methods. HC2 detected HPV in 12% specimens. L1- and E6/E7-PCRs showed 50% sensitivity and 77% specificity.The agreement rate between HC2 and both PCR assays was 65%. Kappa value showed moderate concordance between HC2 and both PCR methods (κ=0.55; CI 95%). Also moderate concordance was seen when L1- and E6/E7-PCRs were compared (κ=0.48; CI 95%). There was a significant association between the Schiller test and E6/E7-PCR (p=0.006) for HPV infection. An acceptable agreement between all three assays for HPV detection was observed. Nevertheless, different PCR formats need to be further analyzed in order to make the right choice of method for HPV testing.

La infección con VPH de alto riesgo es el principal factor etiológico asociado al desarrollo de carcinogénesis cervical y las pruebas de detección de ADN-VPH han mostrado ser una herramienta esencial para la pesquisa y seguimiento de estas infecciones. El objetivo del estudio ha sido comparar tres métodos para la detección del ADN viral, en combinación con los análisis colposcópico y citológico. Se obtuvieron muestras cervicales de 100 mujeres sexualmente activas, en Mérida, Venezuela. La detección de infecciones por VPH se realizó por Captura Híbrida 2 (CH2) y los ensayos de PCR “L1-Nested-PCR” y “E6/E7-PCR”. 40% de las muestras (40/100) fueron positivas para VPH por al menos uno de los métodos aplicados. 12% de las muestras analizadas fueron positivas para VPH por CH2. Las dos PCR utilizadas mostraron un 50% de sensibilidad y 77% de especificidad. La coincidencia observada entre CH2 y las dos PCR fue del 65%. La determinación del valor Kappa mostró una concordancia moderada entre CH2 y ambos métodos de PCR (κ=0,55; CI 95%). También existió concordancia moderada al comparar las PCR de las regiones L1 y E6/E7 de VPH (κ=0,48; CI 95%). Hubo una asociación significativa entre el resultado del test de Schiller y la PCR E6/E7 (p=0,006) para la infección por VPH. Se determinó una concordancia aceptable entre los tres métodos aplicados para la detección de VPH; sin embargo, las PCR deben ser analizadas en trabajos futuros con el fin de establecer las pruebas más adecuadas para la detección viral.

Immunohistochemical staining of Langerhans cells in HPV-positive and HPV-negative cases of oral squamous cells carcinoma

The Human Papillomavirus (HPV) has been strongly implicated in development of some cases of oral squamous cell carcinoma (OSCC). However, the immunological system somehow reacts against the presence of this virus. Among the cells involved in such mechanism of defense Langerhans cells (LC) stand out, which are responsible for processing and presenting antigens. OBJECTIVES: The purposes of this study were to investigate the presence of HPV DNA and to evaluate the immunohistochemical reactivity for Langerhans cells between HPV-positive and HPV-negative OSCC. Twenty-seven cases of OSSC were evaluated. MATERIAL AND METHODS: DNA was extracted from paraffin-embedded tissue samples and amplified by Polymerase Chain Reaction (PCR) for the detection of HPV DNA. Viral typing was performed by dot blot hybridization. Immunohistochemistry was performed by the Streptavidin-biotin technique. RESULTS: From the 27 cases, 9 (33.3 percent) were HPV-positive and 18 (66.0 percent) HPV-negative. HPV 18 was the most prevalent viral type (100 percent cases) and infection with HPV-16 (co-infection) was detected in only 1 case. In the OSCC specimens examined, immunoreactivity to S-100 antibody was detected in all cases, with a mean number of 49.48±30.89 Langerhans cells positive for immunostaining. The mean number of immunostained Langerhans cells was smaller in the HPV-positive cases (38 cells/case) than in the HPV-negative cases (42.5 cells/case), but this difference was not significant (p=0.38). CONCLUSIONS: The low frequency of detection of HPV DNA in OSCC indicates a possible participation of the virus in the development and progression of only a subgroup of these tumors. There was no association between the immunohistochemical labeling for Langerhans cells (S-100+) and HPV infection of in OSSC. These findings suggest that the presence of HPV in such OSCC cases could not alter the immunological system, particularly the Langerhans cells.

Prevalence and multiplicity of HPV in HIV women in Minas Gerais, Brazil / Prevalência e multiplicidade do HPV em mulheres infectadas pelo HIV em Minas Gerais

OBJECTIVE: To detect the frequency and subtypes of HPV in the uterine cervix of HIV-infected women. METHODS: Sample consisted of 288 HIV-infected women, recruited from the public health system of five cities of Minas Gerais, Brazil. Women were seen from August 2003 to August 2008. Cervical samples were collected for cytological analysis and for HPV DNA detection, using polymerase chain reaction (PCR). HPV DNA was classified according to its oncogenic potential in low risk (types 6, 11) and high risk (types 16, 18, 31, 33, 35). Colposcopy was performed, followed by cervical biopsy when necessary. Categorical variables were compared using the Chi-squared test, with a significance level established at the 5 percent level. RESULTS: HPV prevalence was 78.8 percent. Most frequent genotypes were HPV-6 (63.9 percent) and HPV-16 (48.5 percent). High-risk HPV were observed in 70.5 percent of the women; low-risk in 71.4 percent; both high and low-risk HPV were detected in 55.1 percent of the patients. Multiple HPV genotypes were detected in 64.8 percent of the patients; two genotypes in 23.8 percent, and three in 18.9 percent. CONCLUSION: HPV prevalence was high among HIV-infected women. Multiple HPV genotypes were common in samples from the uterine cervix of HIV-infected women.

OBJETIVO: Detectar a frequência e os subtipos do HPV na cérvice uterina de mulheres infectadas pelo HIV. MÉTODOS: A amostra era composta por 288 mulheres infectadas pelo HIV, recrutadas do sistema público de saúde de cinco cidades de Minas Gerais, Brasil. As mulheres foram avaliadas de agosto de 2003 a agosto de 2008. Amostras cervicais foram coletadas para análise citológica e para detecção do HPV DNA, usando a reação em cadeia de polimerase (PCR). O HPV DNA foi classificado de acordo com seu potencial oncogênico em baixo risco (tipos 6,11) e alto risco (tipos 16, 18, 31, 33, 35). Foi realizada colposcopia, seguida de biópsia cervical, quando indicada. Variáveis categóricas foram comparadas usando o teste do quiquadrado, com nível de significância estabelecido de 5 por cento. RESULTADOS: A prevalência do HPV foi 78,8 por cento. Os genótipos mais frequentes foram HPV-6 (63,9 por cento) e HPV-16 (48,5 por cento). Genótipos de HPV de alto risco foram observados em 70,5 por cento das mulheres; de baixo risco em 71,4 por cento; HPV de alto e baixo risco foram detectados em 55,1 por cento das pacientes. Múltiplos genótipos de HPV foram detectados em 64,8 por cento das pacientes; dois genótipos em 23,8 por cento, e três em 18,9 por cento. CONCLUSÃO: A prevalência do HPV foi alta entre mulheres infectadas pelo HIV. Múltiplos genótipos de HPV foram comuns em amostras da cérvice uterina destas mulheres.

Detección de virus papiloma humano (HPV) a partir de muestras celulares de cuello uterino en base líquida: Correlación con la inmunorreactividad de la proteína p16INK4a / Detection of human papilloma virus (HPV) in liquid-based cervical samples: Correlation with protein p16INK4a expression

La citología del cuello uterino en base líquida mejora la calidad de la muestra y el material residual podría ser utilizado para realizar pruebas complementarias, como la detección del virus papiloma humano (HPV) y estudio inmunocitoquímico de biomarcadores. El propósito de este estudio fue correlacionar la presencia de HPV y la inmunoexpresión de p16INK4a en las muestras citológicas en base líquida para examinar la utilidad de estas nuevas herramientas en la detección de cáncer de cuello uterino. Las pacientes incluidas (n=67) presentaban una citología anormal o patología cervical previa. La detección y genotipificación de HPV se realizó con PCR-SPF10/LiPA (INNOLiPA Extra Amp) y para la inmunodetección de p16INK4a se utilizó el anticuerpo clon E6H4. La citología convencional proporcionó los mismos hallazgos citológicos que la citología en base líquida. La prevalencia general del HPV fue de 43,3% (29/67). El HPV16 fue el tipo viral mas frecuente (31,03%) y el 48,3% de los casos presentó infección múltiple. En el 35,8% de las muestras cervicales se detectó inmunoexpresión de la p16INK4a y ésta fue significativamente (p<0,020) asociada a la presencia de HPV. Estos resultados apoyan la evidencia que la implementación de nuevas tecnologías en la rutina diaria del laboratorio contribuye significativamente en la detección precoz del cáncer de cuello uterino y en el aporte de datos importantes para facilitar en el manejo clínico adecuado de la paciente. La detección de HPV combinada con la p16INK4a podría ser utilizado en la evaluación de pacientes con mayor riesgo a desarrollar lesiones cervicales significativas.

The liquid-based cervical cytology improves the quality of the sample and the residual sample could be used efficiently to carry out complementary tests, such as the detection of HPV DNA and the immunocytochemical biomarkers study. The purpose of this study was to correlate the presence of HPV and immunoexpression of p16INK4a in liquid-based cervical samples to examine the utility of these new tools in the detection of cervical cancer. The included patients (n=67) presented an abnormal cytology or previous cervical pathology. The HPV detection and genotyping were carried out with PCR-SPF10/LiPA (INNOLiPA Extra Amp) and for p16INK4a immunodetection was used antibody clone E6H4. The conventional cytology provided the same cytologic interpretations that those of liquid-based cytology. The overall HPV prevalence was 43.3% (29/67). HPV16 was the most frequent viral type (31.03%) and 48.3% of the cases were infected with multiple HPV types. p16INK4a immunoexpression was observed in 35.8% of liquid-based cytological samples and this was significantly (p < 0.020) associated to the HPV presence. These results support the evidence that the implementation of new technologies in the daily routine of the laboratory, contribute significantly in the early detection of cervical cancer and provide important data to help in the patient’s efficient management. The combined use of HPV detection and p16INK4a expression could be used for evaluation of patients with more risk to develop significant cervical lesions.

Genotipificación del virus papiloma humano en mujeres con adenocarcinoma cervical de la Región de La Araucanía-Chile / Human papillomavirus genotyping in cervical adenocarcinoma in the Region of La Araucanía-Chile

El virus papiloma humano (VPH) es el principal factor causal del cáncer cervicouterino (CCU). Así, detectar y genotipificar el VPH es importante para conocer la frecuencia de los genotipos presentes en la región. En este trabajo se estudiaron 44 biopsias de adenocarcinoma cervical (ACC). Para la detección del VPH se empleó una reacción de polimerasa en cadena anidada dirigida al gen L1 (RPCL1), para la genotipificación viral se utilizaron enzimas de restricción (Rsa I, Dde I, Pst I) y secuenciación. Se detectó ADN viral mediante RPCL1 anidada en 100 por ciento de las biopias. Se logró tipificar 38/44 casos: 81,6 por ciento VPH 16; 13,2 por ciento VPH 18; 2,6 por ciento VPH 33 y 2,6 por ciento VPH 18/33. Conclusiones: La metodología fue exitosa para identificar el tipo viral en 86 por ciento de las biopsias. Se observó una estrecha asociación ACC-VPH, especialmente con el tipo viral 16, detectado en 81,6 por ciento de los casos tipificados.

Human papillomavirus (HPV) is the main cause of cervical cancer. Thus, HPV detection and typing becomes important in order to know the frequency of genotypes present in the region. In this paper we studied 44 biopsies of cervical adenocarcinoma. For HPV detection nested polymerase chain reaction (PCR) was used to amplify the L1 gene. For viral typing restriction enzymes (Rsa I, Dde I, Pst I) and DNA sequencing were used. Viral DNA was detected by nested L1 PCR in 100 percent of biopsies; 38/44 cases could be typed: 81.6 percent HPV16; 13.2 percent HPV 18; 2.6 percent VPH 33 and 2.6 percent HPV 18/33. Conclusions: The technique was successful in identifying the virus type in 86 percent of biopsies. There was a strong association ACC-HPV, especially with the viral type 16, detected in 81.6 percent of established cases.

Papilomavírus humano: prevalência e genótipos encontrados em mulheres HIV positivas e negativas, em um centro de referência no extremo Sul do Brasil / Human papillomavirus: prevalence and genotypes found among HIV-positive and negative women at a reference center in the far south of Brazil

INTRODUÇÃO: O objetivo deste estudo foi detectar a presença do papilomavírus humano e verificar a prevalência e distribuição dos genótipos HPV-6, -11, -16 e -18 em mulheres HIV-1 positivas e negativas. MÉTODOS: Analisou-se amostras de secreção cervical de 98 mulheres por reação em cadeia da polimerase nested para presença do HPV e tipo-específica para detecção dos genótipos, sendo estes confirmados por análise dos fragmentos de restrição. Realizou-se os testes do qui-quadrado e Fisher para a análise estatística. RESULTADOS: O DNA-HPV foi observado em 66,3 por cento das amostras analisadas, 76,4 por cento no grupo HIV positivo e 60 por cento no grupo HIV negativo (p=0,1). Uma prevalência maior de infecção viral por genótipos oncogênicos foi observada no grupo de pacientes HIV positivo (65,2 por cento) quando comparado ao grupo HIV negativo (28,6 por cento), (p=0,006), sendo HPV-16 foi o mais frequente nos dois grupos, seguido pelo HPV-18. CONCLUSÕES: Sugere-se que mulheres HIV positivas apresentam maior probabilidade de se infectar por genótipos oncogênicos de HPV, ressaltando a importância de um programa de rastreamento e diagnóstico diferenciado para este grupo.

INTRODUCTION: The aim of this study was detect the presence of human papillomavirus and investigate the prevalence and distribution of the HPV-6, 11, 16 and 18 genotypes among HIV-1 positive and negative women. METHODS: Samples of cervical smears from 98 women were analyzed using the nested polymerase chain reaction for the presence of HPV and using the type-specific technique for genotype detection. Genotypes were confirmed by means of restriction fragment analysis. The chi-square and Fisher tests were performed for the statistical analysis. RESULTS: HPV-DNA was observed in 66.3 percent of the samples analyzed: 76.4 percent in the HIV-positive group and 60 percent in the HIV-negative group (p = 0.1). Higher prevalence of viral infection due to oncogenic genotypes was observed in the HIV-positive group (65.2 percent) than in the HIV-negative group (28.6 percent), (p = 0.006), and HPV-16 was the most frequent in the two groups, followed by HPV-18. CONCLUSIONS: It is suggested that HIV-positive women present a higher likelihood of infection by oncogenic HPV genotypes, and the importance of a screening and differential diagnosis program for this group is highlighted.

High prevalence and low E6 genetic variability of human papillomavirus 58 in women with cervical cancer and precursor lesions in Southeast Mexico

Infection with some genotypes of human papillomavirus (HPV) is the most important risk factor associated with cervical cancer (CC). Throughout the world, HPV type 58 prevalence varies from one region to another; it is higher in women from certain countries in Asia and Latin America, such as China and Mexico. Although intratypic variants have been reported on a few occasions, our knowledge about HPV 58 genetic variation remains limited. Therefore, this work aims to (i) determine the prevalence of HPV type 58 amongst Mexican women with invasive CC or precursor lesions and (ii) identify HPV 58 sequence variants. One hundred and forty five colposcopy clinic patients were studied. Genotyping of HPV 16, 18 and 58 was determined by specific nested PCR and HPV 58 variants were detected by direct sequencing. The general prevalence of HPV was 51.7 percent (75/145). HPV 16 was found in 30.6 percent (23/75) and HPV 58 in 24 percent (18/75) of the patients. HPV 18 was not identified in patients with cervical intraepithelial neoplasia (CIN) grade I; it was only found in those with CIN II, with a prevalence of 6.8 percent (3/44). In patients with CC, the prevalence of HPV 16 and 58 was 78.9 percent. Regarding HPV 58 variants, 94.4 percent of the HPV 58 sequences were identical to the prototype strain, whereas one sample showed changes at a single nucleotide. This study demonstrates a high prevalence of HPV 58 and a low genetic variability of E6 sequences amongst Mexican colposcopy patients.

Human papillomavirus genotypes in asymptomatic young women from public schools in Rio de Janeiro, Brazil / Genótipos de papilomavírus humanos em mulheres jovens assintomáticas de escolas públicas do Rio de Janeiro, Brasil

INTRODUCTION: The aim of this work was to survey HPV information from a random population of young women from Rio de Janeiro, Brazil. METHODS: This cross-sectional study included cervical samples from 241 female students. To determine human papillomavirus status, polymerase chain reaction amplification was performed. HPV typing was determined by restriction fragment length polymorphism analysis. Demographic data, life style, sexual and gynecological history were obtained through use of a structured questionnaire. RESULTS: The average age of the women was 19.6 years-old (SD=3.4 years). HPV prevalence was 27.4 percent. Nineteen different HPV genotypes were detected, including 13 high risk types. HPV 16 was the most prevalent type (6.2 percent), followed by 31 (4.1 percent) and 66 (3.7 percent). Most of the oncogenic types belonged to the A9 species (28/48). The frequency of women infected by at least one oncogenic type was significantly higher than those only infected by low risk types (18.7 percent versus 7.5 percent). Cervical changes were detected in 12.5 percent of the sample and were significantly linked to infection with HPV types of the A9 species. Demographic variables, sexual initiation, or number of sexual partners were not associated with HPV prevalence, variety of HPV genotypes or oncogenic types. CONCLUSIONS: The relative frequency of HPV genotypes other than vaccine types in young females should be taken into account when evaluating vaccination strategies. Due to the high prevalence of HPV infection among the population studied, implementation of sex education in schools, promotion of condom use and an organized screening program to prevent cervical cancer must be encouraged for this age group.

INTRODUÇÃO: O objetivo deste trabalho foi investigar a infecção por HPV em uma população randômica de mulheres jovens no estado do Rio de Janeiro, Brasil. MÉTODOS: Este estudo incluiu amostras cervicais de 241 jovens. A detecção de papilomavírus humanos foi realizada pela reação da cadeia da polimerase e a tipificação deste vírus determinada pelo polimorfismo do comprimento do fragmento de restrição. Dados demográficos, estilo de vida, história sexual e ginecológica foram obtidos através de um questionário. A média de idade das mulheres foi 19,6 anos (SD=3,4). RESULTADOS: A prevalência de HPV foi de 27,4 por cento e 19 diferentes tipos foram detectados, incluindo 13 oncogênicos. O HPV 16 foi o tipo mais prevalente (6,2 por cento), seguido do 31 (4,1 por cento) e do 66 (3,7 por cento). A maioria dos tipos oncogênicos pertencia à espécie A9 (28/48). A frequência de mulheres infectadas por ao menos um tipo oncogênico foi significantemente mais alta do que aquelas infectadas por tipos de baixo risco (18,7 por cento versus 7,5 por cento). Alterações cervicais foram detectadas em 12,5 por cento das jovens e foram positivamente relacionadas a tipos de HPV da espécie A9. Variáveis demográficas, iniciação sexual e número de parceiros sexuais não foram associados com a prevalência de HPV, variedade de genótipos ou tipos oncogênicos. CONCLUSÕES: A frequência razoável de outros genótipos além dos vacinais nestas jovens deve ser considerada para avaliação de estratégias de vacinação. Devido à alta prevalência de HPV na população estudada, a implantação de educação sexual nas escolas, incentivo ao uso de preservativo e exames preventivos devem ser encorajados nesta faixa etária.

Prevalence of human papillomavirus (HPV) and HPV-16 genotyping by real-time PCR in patients with several cervical pathologies

PURPOSE: this study was planned to evaluate the prevalence of HPV (excepting type 16) and HPV 16 by real-time PCR in colposcopy patients and to interprete the results with age, age of first sexual intercourse (FSI), parity and Pap smear results. METHODS: one hundred and two colposcopy patients (50 and 52 of the patients were classified as colposcopy positive and negative, respectively) applying to Gynecology clinic were included. HPV (excepting type 16) and HPV 16 were detected by realtime PCR using the L1 region. Real-time nested amplifications of MY09/11 products were done by GP5+/GP6+ primers and Cyanine-5 labeled HPV and HPV 16 DNA specific probe after HPV DNA extraction by phenol chloroform isoamylalcohol. RESULTS: HPV (excepting type 16) and HPV 16 were positive in 12 percent and 18 percent of the colposcopy positive patients respectively. HPV (excepting type 16) and HPV 16 were positive in 5.7 percent and 3.8 percent of the colposcopy negative patients, respectively. CONCLUSION: there was a statistically significant difference between colposcopy positive and colposcopy negative patients comparing HPV 16 with total HPV positivity (p = 0.021 for type 16 and p = 0.010 for total HPV) but there was not a statistically significant difference between colposcopy positive and colposcopy negative patients when we compared HPV (excepting type 16) positivity (p = 0.314). In conclusion, HPV detection and typing may be helpful for cervical cancer screening and prevention.

Genotipos de virus papiloma humano (VPH) en pacientes con cáncer cervico-uterino en un hospital público y una clínica privada de Santiago, Chile / Human papillomavirus (HPV) genotypes in cervix uterine cancer patients in a public hospital and private clinic from Santiago, Chile

We compared HPV genotypes among squamous cervical cancer samples from a public hospital (n = 55) and a private clinic (n = 35 cases) of Santiago. Paraffin-embedded specimens were analyzed by PCR followed by an immunoenzimatic assay. Reverse line blotting was used for the identification of 36 HPV genotypes. We found HPVDNAm 94.4 percent of all cancers. Single mfections: HPV16: 40.0 percent, (clinic 37.1 percent, hospital 41.8 percent) VPH18:7.8 percent (clinic 2.9 percent, hospital 10.9 percent); single+multiple mfections: VPH16: 61.1 percent (clinic 53.1 percent, hospital 71.7 percent), VPH18: 34.4 percent (clinic 21.9 percent, hospital 45.2 percent). HPV16 orHPV18 occurredin 75.6 percent of cases, higher inthe hospital than the clinic (87.3 percent-95 percent CI: 84.9-96.3 - and 57. l percent-95 percent CI: 46.6-66 - respectively, p = 0.002). Other genotypes in single mfections: HPV 26, 31, 33, 45, 58, 67; in co-mfections: HPV 35,52,56,59 and 66. HPV16 but specially HPV 18 were significantly more frequent in the public hospital; 75.6 percent of squamous cervical cancer were associated to the vaccine preventable HPV16/18.

Se comparan los genotipos de VPH en casos de cáncer cérvico-uterino escamocelular de una clínica privada (n: 35) y de un hospital público (n: 55) atendidos entre 1996 y 2006 en Santiago, Chile. Se analizaron por RPC y ensayo inmunoenzimático muestras tumorales en bloques de parafina, genotipificándose con reverse Une blotting para 36 genotipos de VPH. Se detectó VPH en 94,4 por ciento de los casos: infecciones únicas por: VPH 16: 40,0 por ciento>, (clínica 37,1 por ciento, hospital 41,8 por ciento) VPH 18: 7,8 por ciento (clínica 2,9 por ciento, hospital 10,9 por ciento); total de infecciones por VPH 16 61,1 por ciento (clínica 53,1 por ciento, hospital 71,7 por ciento), por VPH 18 34,4 por ciento (clínica 21,9 por ciento, hospital 45,2 por ciento). Co-infección: VPH 16/18 75,6 por ciento (clínica 57,1 por ciento; IC95 por ciento = 46,6-66,0 hospital 87,3 por ciento; IC95 por ciento = 84,9-96,3, p = 0,002). Se identificó otros 11 genotipos oncogénicos en infecciones únicas (VPH: 26, 31, 33, 45, 58, 67) o en co-infección con VPH 16/18 (VPH: 35, 52, 56, 59, 66). VPH 16 y VPH 18 fueron significativamente más frecuentes en el hospital público, particularmente VPH18; 75,6 por ciento> de los cánceres se asociaron a los genotipos VPH 16/18, tipos prevenibles por vacuna.

Risk factors associated with human papillomavirus infection in two populations from Rio de Janeiro, Brazil

We investigated human papillomavirus (HPV) infection in two female populations from diverse socio-economic strata from the state of Rio de Janeiro and we also investigated the possible co-factors related to infection and the progression to cancer. In Group I, the reference group of this study, 10.7 percent of the patients presented HPV infection, as detected by generic PCR, while in Group II (low socio-demographic conditions) HPV was detected in 31.1 percent of the samples. HPV16 was the most prevalent virus type found in both Groups I and II (5.3 percent and 10 percent, respectively), followed by HPV 18 (1.3 percent and 4.7 percent, respectively). Although only a small sample was analysed, we detected differences among the groups regarding the rates of HPV infection, HPV types, age, ethnicity, familial income, schooling, marital status, parity, tobacco smoking and oral contraceptive use. For Group I, the Papanicolaou test was the most powerful independent factor associated with HPV status, followed by an age of under 30 years old, the number of sexual partners and black ethnicity. Our data are in agreement with the co-factors that are typically described for the developed world. For Group II, the Pap test was also the most relevant variable that was analysed, but the history of other sexually transmitted diseases and the use of alcohol were additional factors that were implicated in infection. These findings point out the need for the development of general and specific strategies for HPV screening of all Brazilian women.