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.

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.

Análise crítica dos métodos moleculares para detecção do papilomavírus humano: revisão da literatura / Molecular methods for human papillomavirus detection: literature review

O papilomavírus humano (HPV) é um vírus que infecta o epitélio cutâneo e das mucosas. Células infectadas por este vírus perdem a capacidade de controlar o ciclo celular e passam a proliferar descontroladamente gerando alterações displásicas que podem progredir para lesões malignas. Existem mais de 200 tipos de HPV e entre eles aqueles que apresentam maior ou menor risco de causar câncer. Dessa forma, o HPV pode ser classificado como sendo de baixo risco ou alto risco. Os métodos mais utilizados em pesquisa para análise molecular do HPV é a hibridização in situ (ISH), reação em cadeia da polimerase (PCR) que varia desde a PCR alelo específica, passando pelo tipo Nested e a PCR multiplex, e a mais nova técnica baseada na tecnologia de microarray. A maioria destes testes é realizada apenas em centros de pesquisa, e não rotineiramente na clínica. Por outro lado, o teste de captura híbrida II para HPV, baseado na hibridização do DNA, é comercialmente disponível. As técnicas para detecção do DNA do HPV e sua genotipagem variam quanto a sua sensibilidade e especificidade. Técnicas que utilizam sondas como a hibridização in situ e o Southern blotting são as menos sensíveis para detecção da sequência do DNA, enquanto que as mais sensíveis são as técnicas que utilizam a amplificação do DNA alvo, como a PCR e a qPCR. À medida que a tecnologia avança, as técnicas moleculares vão se aprimorando para a detecção do HPV. O objetivo final é desenvolver uma metodologia de baixo custo que apresente resultados rápidos e eficientes.

The human papillomavirus (HPV) is a virus that infects the skin and mucosal epithelium. Infected cells lost the ability to control cell cycle and begin to proliferate uncontrollably causing dysplastic alterations that can progress to malignant lesions. There are over 200 types of HPV with higher or lower risk of causing cancer. Thereby, HPV can be classified as high risk or low risk. The methods used in research for molecular analysis of HPV is the in situ hybridization (ISH), polymerase chain reaction (PCR) that varies from the allele specific PCR, Nested, PCR multiplex, and the newest technique based on microarray technology. Most of these tests are performed only in research centers, and not routinely in the clinic. An exception is the Hybrid Capture II test for HPV. The detection techniques of HPV and its genotyping vary in their sensitivity and specificity. Techniques that use probes, as in situ hybridization and Southern blotting are less sensitive for detection of DNA sequence, while the most accurate are the techniques based on DNA amplification, such as PCR and qPCR. As technology advances, molecular techniques become more accurate for the detection of HPV. The ultimate goal is to develop an inexpensive method to provide rapid and efficient results.

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.

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.

Detección de Virus Papiloma Humano en el fluido gingival de pacientes con inmunodeficiencia humana y enfermedad periodontal / Detection of Human Papillomavirus in gingival fluid of Venezuelan HIV patients with periodontal disease

Evidencias sugieren que los virus pueden participar en la activación de la enfermedad periodontal, permitiendo el sobrecrecimiento de bacterias periodontopatógenas. El objetivo del estudio fué la detección molecular de VPH en fluido gingival (FG) de pacientes VIH+ con enfermedad periodontal. Se evaluaron muestras de FG de 20 pacientes VIH+ con enfermedad periodontal que asistieron al Centro de Atención de Pacientes con Enfermedades Infecciosas (CAPEI) de la Facultad de Odontología de la Universidad Central de Venezuela, 13 bajo terapia antirretroviral (HAART) y 7 VIH+ sin HAART. Se incluyeron 7 pacientes seronegativos con periodontitis crónica y como grupo control 7 pacientes seronegativos periodontalmente sanos. Se extrajo el ADN, se amplificó la región L1 de VPH con primers MY09 y MY11. Las muestras VPH+ fueron genotipificadas para los tipos 6, 11, 16, 18, 31 y 45. VPH fue detectado en 46% de los pacientes VIH+ bajo terapia. El contaje CD4+ en la población VPH+ no presentó diferencias con el grupo VPH-, y la carga viral mostró valores promedio significativamente mayores (200.470± 324.244 copias/mL) con respecto a los pacientes VPH- (10.246±23.805 copias/mL). Las muestras VPH+ presentaron los genotipos 6 y 11, de los cuales 66,6% estaban coinfectados con ambos tipos. Las condiciones periodontales no presentaron diferencias entre los individuos con doble infección viral por VPH y VIH, y los que solo portaban VIH. VPH fue detectado solamente en fluido gingival de pacientes VIH+ con HAART, indicando que esta terapia puede influir en el estado inmunológico independientemente de las condiciones periodontales.

Evidence suggests that viruses may be involved in the activation of periodontal disease, allowing the overgrowth of periodontal pathogens. The purpose of the present study was to detect the presence of Human Papillomavirus (HPV) in gingival crevicular fluid (GCF) in HIV+ Venezuelan patients with periodontal disease. We evaluated GCF samples from 20 HIV+ patients with periodontal disease from the Infectious Disease Center, Faculty of Dentistry, Central University of Venezuela, and were clinically examined to establish their periodontal conditions, 13 under HAART (antiretroviral therapy) and 7 without HAART. Seven seronegative patients with chronic periodontitis and 7 seronegative patients, without periodontal disease were included. DNA extraction was performed, the consensus primers MY09 and MY11 for the HPV L1 region were used for PCR amplification. Genotipification was made for the 6, 11, 16, 18, 31 and 45 genotypes. HPV were detected in 46% of HIV+ patients under therapy. The CD4 cell counts in the HPV+ patients were not significantly different from the HPV-group. The viral load in the HPV+ group was significantly higher (200,470 ± 324,244 copy/mL) than in the HPV- patients (10,246 ± 23,805 copy/mL). Genotypes 6 and 11 were observed in the HPV positive samples, of which 4/6 (66.6%) presented coinfection with both types. No significant differences in the periodontal conditions were observed between patients with HPV-HIV infection related to patients with only HIV. HPV was detected only in the gingival crevicular fluid of HIV+ patients under HAART independently of the periodontal conditions.

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.

Short-term risk of cervical intraepithelial neoplasia grades 2 and 3 for women with normal cytology and human papillomavirus infection / Riesgo a corto plazo de lesiones intraepiteliales cervicales grados 2 y 3 en mujeres con citología vaginal normal e infección por el virus del papiloma humano

OBJECTIVE. To assess the risk of cervical intraepithelial neoplasia grades 2, 3 or higher (CIN 2/3+) for women with normal cytology and concurrent high-risk human papillomavirus infection (HR-HPV). MATERIAL AND METHODS. We examined 2 200 women every 6 months for an average of 9 years. Cervical smears and samples for HPV DNA were obtained at each visit. Absolute risk of subsequent CIN2/CIN3+ was estimated using the Kaplan-Meier method. RESULTS. The absolute risk of CIN2/CIN3+ among HR-HPV-positive women with normal Pap smear results was 1.06 percent (95 percentCI, 0.57-2.20), 5 times higher the risk among all women with normal Pap smears (0.20 percent; 95 percentCI, 0.12-0.32) but 7 times lower than that for women with HR-HPV infection and LSIL (7.24 percent; 95 percentCI, 3.78-15.2). CONCLUSION. Short-term absolute risk of CIN2/3+ after a normal Pap smear with concurrent HR-HPV infection is low (~1 percent), suggesting that the HR-HPV test has limited utility in short-term clinical decision-making for women with normal cytology.

OBJETIVO. Evaluar el riesgo a corto plazo de neoplasia intraepitelial cervical de alto grado (CIN2/CIN3+) en mujeres con citologí-a cervicouterina normal e infección por virus del papiloma humano de alto riesgo (HR-HPV). MATERIAL Y MÉTODOS. Cohorte prospectiva de 2200 mujeres evaluadas cada seis meses durante 9 años en promedio. En cada visita se tomó muestra cervical para extendido y detección de HPV DNA. El riesgo absoluto de CIN2/CIN3+ a la siguiente visita fue calculado utilizando el método de Kaplan-Meier. RESULTADOS. En mujeres con citologí-a normal e infección concomitante por HR-HPV el riesgo absoluto de presentar CIN2/CIN3+ fue de 1.06 por ciento (95 por cientoCI, 0.57-2.20). Este riesgo fue cinco veces mayor al observado en todas las mujeres con citologí-a normal (0.20 por ciento; 95 por cientoCI, 0.12-0.32) pero siete veces menor que el observado en mujeres con lesiones intraepiteliales escamosas de bajo grado con infección concomitante (7.24 por ciento; 95 por cientoCI, 3.78-15.2). CONCLUSIÓN. El riesgo absoluto de CIN2/3+ a corto plazo luego de una citologí-a normal e infección por HR-HPV es baja (~1 por ciento), sugiriendo que, a corto plazo, la prueba de HR-HPV tiene utilidad clí-nica muy limitada en mujeres con citologí-a normal.

Nuevos paradigmas y desafíos en la prevención y control del cáncer de cuello uterino en América Latina / New paradigms and challenges in cervical cancer prevention and control in Latin America

El cáncer de cuello uterino 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, 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 VPH-16 y VPH-18 en mujeres no expuestas previamente al virus representan una gran oportunidad para la prevención del cáncer de cuello uterino en la región. En este manuscrito resumimos la evidencia científica y la experiencia de la región en i) el uso de pruebas de VPH y de la inspección visual después del ácido acético (IVAA) en tamizaje primario, y ii) la implementación de programas de vacunación en adolescentes. Finalmente enumeramos una serie de recomendaciones adecuadas para distintos escenarios. La factibilidad de implementar un programa nacional de prevención de cáncer de cuello uterino exitoso y sostenible en países latinoamericanos dependerá de las prioridades de salud, la infraestructura y personal de salud disponible, determinadas luego de un riguroso análisis situacional local.

Cervical cancer continues to be a significant health problem in Latin America. The use of conventional cytology to detect precancerous cervical lesions has had almost no major impact on reducing cervical cancer incidence and mortality rates, which are still high in the region. The availability of new screening tools to detect precancerous lesions provide great opportunities for cervical cancer prevention in the region, as do highly efficacious HPV vaccines able to prevent nearly all lesions associated with HPV-16 and -18 when applied before viral exposure. This paper summarizes the scientific evidence and regional experiences related to: i) the use of HPV testing and visual inspection after the application of acetic acid (VIA) in primary screening and ii) the implementation of adolescent HPV vaccination programs. Finally, we outline a number of recommendations for different resource settings. The feasibility of implementing successful and sustainable national cervical cancer prevention programs in Latin American countries in the region will depend on health priorities and the availability of infrastructure and health personnel-as determined by rigorous local situational analysis.

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.

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.

Oral HPV infection in a bone marrow transplantation patient: a case report with atypical clinical presentation and unexpected outcome

HPV (Human Papilloma Virus) is one of the most prevalent infections worlwide. Oral HPV infection may be associated with different diseases of oral cavitie. Although oral HPV infection occurs frequently, it rarely causes lesions. An increased rate of oral HPV-induced lesions is observed in people with an impaired immune system. The most common conditions induced by oral HPV infection are focal epithelial hyperplasia, oral condylomas and oral papillomas. We reported a case of oral HPV lesion in a bone marrow transplantation patient with atypical clinical presentation and unexpected outcome.

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.

A frequência do HPV na mucosa oral normal de indivíduos sadios por meio da PCR / The frequency of human papillomavirus findings in normal oral mucosa of healthy people by PCR

Os papilomavírus humanos (HPV) são DNA vírus pertencentes à família papilomaviridae com grupos de baixo e alto risco que infectam a pele e a mucosa podendo induzir a formação de tumores epiteliais benignos e malignos. Na mucosa oral, estes vírus têm sido associados a papilomas orais, hiperplasias epiteliais focais, leucoplasias e neoplasias orais. Objetivo: Estudar a frequência do HPV em mucosa oral de indivíduos normais. Material e método: Trabalho prospectivo em coorte transversal. Participaram desse estudo 100 indivíduos voluntários, faixa etária de 20 a 31 anos, estudantes universitários, sem história, queixas ou lesões visíveis ao exame físico de cavidade oral e orofaringe. Foram submetidos a questionário com perguntas referentes à epidemiologia da infecção pelo HPV. Foi colhido material de mucosa oral por raspado com escova e analisado pelo PCR. Resultados: Os resultados mostraram ausência de HPV em todas as amostras. Conclusão: Parece ter havido participação do alto nível socioeconômico com alimentação rica em carotenoides e vitamina C, baixo consumo tabágico e etílico e comportamento heterossexual predominantemente monogâmico com uso regular de preservativos.

The human papillomavirus (HPV) is a DNA virus, which belongs to papillomaviridae family, being of low and high risk, which infect the skin and mucous membranes and can induce benign and malign tumor formation. In the oral mucosa they have been associated with oral papilloma, focal epithelial hyperplasia, leucoplakia and oral neoplasia. AIM: to study the frequency of HPV finding in oral mucosa of normal people. Materials and methods: Prospective study, cross-sectional cohort. One hundred volunteers, young adults, healthy, aged between 20 and 31 years, university students with no history, no complains, without oral or oropharyngeal lesions. They were submitted to a questionnaire with questions regarding HPV infection epidemiology. The samples were harvested by brushing and analyzed by PCR. Results: The results were negative for HPV in all samples. Conclusion: It seems we had high social and economical class individuals, with nutrition rich in carotenoyds and vitamin C, low smoking and alcohol consumption and heterosexual habits with predominant monogamy and regular use of condoms.

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.

AgNOR polymorphism association with squamous intraepithelial lesions and invasive carcinoma with HPV infection / Asociación de los polimorfismos AgNORs con lesiones intraepiteliales escamosas, carcinoma cervical e infección por VPH

OBJECTIVE: Evaluate the relationships between AgNORs polymorphisms and squamous intraepithelial lesions (SIL) and squamous cell carcinoma (SCC) with HPV infection. MATERIALS AND METHODS: A study was carried out on sixty women from the state of Guerrero, Mexico. HPV detection was performed by PCR. AgNORs were identified by argentic impregnation. One hundred cells per slide were counted and classified according to the polymorphism of AgNORs dots; typical (spherical) and atypical (large, kidney-shaped and clustered). RESULTS: A total of 100 percent of the cases were positive for HPV infection. Nine different high-risk HPV genotypes were found, type16 was the most common (48.6 percent). The AgNORs showed a significant decrease in spherical shape according to neoplastic development. The three atypical shapes showed a significant increase in SIL and SCC (p-trend<0.001). CONCLUSIONS: AgNORs polymorphism rises progressively according to the grade of histological lesions that can be useful as a prognosis for progression of SCC.

OBJETIVO: Evaluar la relación entre los polimorfismos de AgNORs con las lesiones intraepiteliales escamosas (LIE) y carcinoma de células escamosas (CCE). MATERIAL Y MÉTODOS: Se estudiaron sesenta mujeres del estado de Guerrero, México. La detección del VPH fue por PCR y los AgNORs por impregnación argéntica; se contaron 100 células y se clasificaron por tipo de polimorfismo de AgNORs: típico (esférico) y atípicos (largo, forma de riñón o de racimo). RESULTADOS: El 100 por ciento de los casos presentaron infección por VPH, se encontraron nueve genotipos diferentes de VPH de alto riesgo, el 16 fue el más común (48.6 por ciento). La forma esférica de los polimorfismos de AgNORs mostró una disminución con el desarrollo neoplásico y las atípicas incrementaron progresivamente con SIL y SCC (p-tendencia<0.001). CONCLUSIONES: Los polimorfismos AgNORs se incrementan progresivamente con el grado de lesión histológica, y pueden ser útiles en el pronóstico de progresión del carcinoma cervical.

Detección y tipificación de virus papiloma humano en adenocarcinoma de cuello uterino mediante reverse line blot, Región de La Araucanía, Chile / Detection and genotyping of human papillomavirus in biopsies of uterine cervical adenocarcinoma

Background: The genotyping of Human Papillomavirus (HPV) will improve knowledge about the local epidemiological association of this virus with adenocarcinoma. Aim: To determine the frequency of HPV genotypes in biopsies of women with uterine cervical adenocarcinoma in a geographic region of Chile. Materials and Methods: Forty-one cervical biopsies with a pathological diagnosis of adenocarcinoma, corresponding to all women diagnosed with this cancer between 2002 and 2004, were analyzed. Viral gene Ll was amplified by PCRfor viral detection. HPV genotyping was carried out by a Reverse Line Blot technique. Results: Seventy one percent of biopsies were positive for HPV. The most common genotypes found were HPV 16 (61 percent), followed by HPV 18 (19.5 percent). Eighty seven percent of biopsies had a single HPV infection. Three patients had a multiple HPV infection. All of the latter were infected by HPV 16, associated with other three viral genotypes (45, 52 and 66). No low-risk HPV genotypes were found. Conclusions: In this sample of biopsies, there was a high prevelence of HPV 16 and a low prevalence of HPV 18, which historically has been related to adenocarcinoma. The genotypes found correspond to those described in South America.

Prevalencia de la infección genital por virus papiloma humano en hombres universitarios voluntarios de la IX Región, Chile / Prevalence of human papillomavirus genital infection among male university students

Background: Human papillomavirus (HPV) infection is the most common sexually transmitted disease. Aim: To determine prevalence of HPV genital infection in voluntary asymptomatic male university students. Material and methods: A cross-sectional study in 62 asymptomatic, sexually active male students. Exfoliated cells were obtained from the penüe shaft and coronal sulcus. Samples were analyzed for HPV DNA detection and genotyping by polymerase chain reaction and Reverse Line Blot. Results: The prevalence of HPV infection was 84 percent. HPV detection was 77 percent in penile shaft and 66 percent in coronal sulcus. The most commonly detected types were HPV-16 (45 percent), HPV-11 (19 percent), HPV-6 (10 percent) and HPV-18 (9 percent). Múltiple infection wasfoundin 54 percent. The most frequent combinations were VPH11/16 (18 percent) and VPH16/18 (5 percent). Conclusions: HPV infection is highly frequent in asymptomatic male university students, high rísk HPV types were greatly predominant.