Papillomavirus infections in the oral and genital mucosa of asymptomatic women

Abstract Human papillomavirus (HPV) has been found in several regions of the body, including the oral cavity. Recently, this virus has been associated with oropharyngeal cancer, but little is known about HPV transmission to the oral cavity. We carried out a study to investigate concurrent oral and cervical infections in 76 asymptomatic women attending a healthcare program. Demographic and behavior data were obtained through a structured questionnaire. Oral and cervical mucosa scrapings were collected and stored for DNA extraction. HPV DNA amplification was performed by polymerase chain reaction assay (PCR) using both primers My09/My11 and FAP59/64, followed by HPV typing with restriction fragment length polymorphism analysis (RFLP) and sequencing. The data collected revealed no risk factors for HPV infection in these 76 women. HPV prevalence of 9.2 and 5.3% was found in cervical and oral mucosa, respectively. Concurrent infections by discordant types were detected in one case only. Sequencing procedures allowed us to detect a new putative HPV 17 subtype from the Betapapillomavirus genus. Our results support the view that cervical and oral HPV infections are independent events. The observed low prevalence of both oral and cervical HPV infections could be associated with attendance in a healthcare program.

Epstein-Barr virus in oral mucosa from human immunodeficiency virus positive patients / Vírus Epstein-Barr na mucosa oral de pacientes positivos para o vírus da imunodeficiência humana

Objective: the detection rate of Epstein-Barr virus (EBV) is higher in people living with human immunodeficiency virus (HIV). In an attempt to contribute to our epidemiological understanding of this coinfection and to investigate the activity of EBV in normal oral mucosa, we performed a cross-sectional study with HIV-positive patients. Methods: oral smears from 145 HIV-positive patients were collected between March 2010 and March 2011. Nested polymerase chain reaction (PCR) and reverse transcriptase-PCR (RT-PCR) were used to genotype EBV and to detect EBNA-2 expression, respectively. Results: EBV DNA was detected in 48.3% of the study participants, of whom 32.85% were EBV-1 and 45.71% were EBV-2 carriers. Additionally, 14.28% were coinfected with both types. EBNA-2 mRNA was expressed in 45.7% of the EBV -positive samples, including 20.0% with EBV-1 only, 20.0% with EBV-2 only and 1.4% with both genotypes. Immune status affected the overall EBV infection, and EBV-2 positivity was significantly correlated with sexual lifestyle of the participants. EBV co-infection with both viral types was dependent upon HIV viral load and the activity of the EBNA-2 gene. Conclusion: we report a high prevalence of active EBV in the oral mucosa of asymptomatic HIV-seropositive individuals. This study addresses the need for monitoring and treatment of HIV-infected patients with EBV reactivation. .

Objetivo: a taxa de detecção do vírus Epstein-Barr (EBV) é alta em pacientes vivendo com o vírus da imunodeficiência humana. Com o objetivo de contribuir para o entendimento epidemiológico e investigar a atividade do EBV na mucosa oral, foi realizado um estudo de coorte com pacientes HIV positivos. Métodos: esfregaços orais de 145 pacientes HIV positivos foram coletados entre março de 2010 e março de 2011. A reação de cadeia de polimerase (PCR) internalizada e a PCR reversa (RT-PCR) foram usadas para genotipar o EBV e detectar a expressão do EBNA-2, respectivamente. Resultados: o DNA do EBV foi detectado em 48,3% dos participantes, dos quais 32,85% eram portadores do EBV-1 e 45,71% de EBV-2. Adicionalmente, 14,28% eram co-infectados por ambos os tipos. O mRNA do gene EBNA-2 foi expresso em 45,7% das amostras positivas para EBV, incluindo 20% por EBV-1 somente, 20% por EBV-2 somente e 1,4% por ambos os genótipos. O estado imune afetou a infecção por EBV, e a positividade para EBV-2 foi significativamente correlacionada com o comportamento sexual dos participantes. A co-infecção por ambos os genótipos de EBV foi dependente da carga viral de HIV e da atividade do gene EBNA-2. Conclusão: registrou-se uma alta prevalência de EBV em atividade na mucosa oral de indivíduos assintomáticos soropositivos para HIV. O estudo focaliza a necessidade de monitoramento e tratamento de pacientes infectados por HIV com reativação pelo EBV. .

Human papillomavirus detection in cervical scrapes from women attended in the Family Health Program / Detecção do papilomavírus humano em citologias cervicais de mulheres atendidas no Programa Saúde da Família / Detección del papilomavirus humano en citologías cervicales de mujeres atendidas en el Programa de Salud de la Familia

OBJECTIVES: to survey the prevalence of human papillomavirus, associated risk factors and genotype distribution in women who were referred to cervical cancer screening when attended in a Family Health Program. METHOD: we conducted a cross-sectional survey, investigating 351 women. Polymerase chain reaction for DNA amplification and restriction fragment length polymorphism analysis were used to detect and typify the papillomavirus. RESULTS: virus infection was detected in 8.8% of the samples. Among the 21 different genotypes identified in this study, 14 were high risk for cervical cancer, and the type 16 was the most prevalent type. The infection was associated with women who had non-stable sexual partners. Low risk types were associated with younger women, while the high risk group was linked to altered cytology. CONCLUSION: in this sample attended a Family Health Program, we found a low rate of papillomavirus infection. Virus frequency was associated to sexual behavior. However, the broad range of genotypes detected deserves attention regarding the vaccine coverage, which includes only HPV prevalent types. .

OBJETIVOS: identificar a prevalência do papilomavírus humano, os fatores de risco associados e a distribuição dos genótipos nas mulheres referenciadas para rastreamento do câncer cervical, durante o atendimento no Programa de Saúde da Família. MÉTODO: foi realizado levantamento de corte transversal com 351 mulheres. A amplificação do DNA por reação da cadeia da polimerase e a análise do polimorfismo de fragmentos de restrição foram aplicadas para detectar e tipificar o papilomavírus. RESULTADOS: a infecção pelo vírus foi detectada em 8,8% das amostras. Entre os 21 genótipos diferentes identificados neste estudo, 14 foram de alto risco para o câncer cervical, e o tipo 16 foi o mais prevalente. A infecção foi associada a mulheres com parceiros sexuais diversos. Os tipos de baixo risco foram associados com mulheres mais novas, enquanto o grupo de alto risco foi vinculado a alterações citológicas. CONCLUSÃO: nesta amostra, atendida por um Programa de Saúde da Família, encontraram-se níveis baixos de infecção pelo papilomavírus. Foi detectada, também, associação entre a frequência do vírus e o comportamento sexual. Porém, a grande variedade de genótipos detectados merece atenção relacionada à cobertura vacinal, que abrange somente tipos prevalentes de HPV. .

OBJETIVOS: identificar la prevalencia del papilomavirus humano, los factores de riesgo asociados y la distribución de los genotipos en las mujeres referenciadas para rastreo del cáncer cervical durante la atención en el Programa de Salud de la Familia. MÉTODO: fue efectuado un levantamiento trasversal con 351 mujeres. La amplificación del DNA por reacción de la cadena de la polimerasa y el análisis del polimorfismo de fragmentos de restricción fueron aplicadas para detectar y tipificar el papilomavirus. RESULTADOS: la infección por el virus fue detectada en el 8,8% de las muestras. Entre los 21 genotipos diferentes identificados en este estudio, 14 fueron de alto riesgo para el cáncer cervical, y el tipo 16 fue el más prevalente. La infección fue asociada con mujeres con parejas sexuales diversas. Los tipos de bajo riesgo fueron asociados con mujeres más nuevas, mientras el grupo de alto riesgo fue simplemente vinculado a alteraciones citológicas. CONCLUSIÓN: en esta muestra atendida por un Programa de Salud de la Familia, encontramos niveles bajos de infección por el papilomavirus. Fue encontrada una asociación entre la frecuencia del virus y el comportamiento sexual. Sin embargo, la gran variedad de genotipos detectados merece atención relacionada a la cobertura de vacunación, que abarca solamente tipos prevalentes de HPV. .

Prevalence of human papillomavirus and Epstein-Barr virus DNA in penile cancer cases from Brazil

Penile cancer is a potentially mutilating disease. Although its occurrence is relatively rare worldwide, penile cancer rates can be high in developing countries. A few studies have been conducted on the involvement of human papillomavirus (HPV) in penile carcinoma, which have found HPV present in 30-70 percent of penile malignant lesions, with a higher prevalence of HPV 16 and 18. It has been assumed that cofactors, such as Epstein-Barr virus (EBV) infections, may play a role in the progression of penile neoplasia. The aim of this study was to determine HPV and EBV prevalence in 135 penile malignant lesions from Brazilian men through the use of MY09/11 polymerase chain reaction (PCR), type-specific PCR and restriction fragment length polymorphism analysis. HPV prevalence among the men tested was 60.7 percent. Of the men who tested positive, 27 presented with HPV 16 (29.7 percent), five with HPV 18 (5.5 percent), 21 with HPV 45 (23.1 percent) and nine with HPV 6 (9.9 percent). Seven mixed infections were detected (9.2 percent), while 11 cases remained untyped (13.4 percent). Regarding EBV positivity, 46.7 percent of the samples contained EBV DNA with EBV-1 as the most prevalent type (74.6 percent). More than 23 percent of the men were co-infected with both HPV and EBV, while 35 percent presented exclusively with HPV DNA and 20 percent presented only with EBV DNA. Penile carcinoma aetiology has not been fully elucidated and the role of HPV and EBV infections individually or synergistically is still controversial. Hence, more studies are needed to determine their possible role in carcinogenesis.

Epstein Barr virus detection in cervical samples of women living with human immunodeficiency virus / Detecção do vírus Epstein-Barr em amostras cervicais de mulheres infectadas pelo vírus da imunodeficiência humana

Epstein Barr Virus (EBV) is transmitted commonly by saliva, but it has been found in genital secretions, which suggests sexual transmission and led researchers to connect EBV and cervical neoplasia. People living with human immunodeficiency virus (HIV) are reported to be at high risk of acquiring genital infections and cervical lesions. To verify the presence of EBV in the genital tract and/or it could affect cervical changes, we analyzed cervical smears from 85 HIV seropositive women for EBV DNA determination. EBV was only detected in two (2.3 percent) samples. The present study provides neither evidence for EBV as sexually transmitted infection nor discards this possibility.

O vírus Epstein-Barr (EBV) é transmitido comumente pela saliva, mas pode ser encontrado também em secreções genitais, sugerindo transmissão sexual e levando pesquisadores a associar este vírus à neoplasia cervical. Pessoas infectadas pelo virus da imunodeficiência humana (HIV) são de alto risco para aquisição de infecções genitais e lesões de cérvice uterina. Com o objetivo de verificar a presença do DNA do EBV no trato genital e/ou se poderia ter efeito em alterações cervicais, analisamos esfregaços cervicais de 85 mulheres HIV soropositivas. O vírus foi detectado em apenas duas (2,3 por cento) amostras. O presente estudo não fornece evidência da transmissão sexual do EBV, nem descarta esta possibilidade.

Update in the human papillomavirus infection prevalence rates and risk fators associated with penile cancer cases / Atualização no conhecimento acerca da prevalência das infecções Causadas pelos papilomavírus humanos e fatores de risco associados aos casos de câncer de pênis

O carcinoma de pênis é uma doença rara e potencialmente mutilante, com etiologia ainda pouco conhecida. A infecção pelo papilomavírus humano (HPV) parece ter um papel importante no desenvolvimento de um subgrupo desses carcinomas e a sua presença parece estar relacionada com determinados tipos histológicos. A prevalência do HPV em tumores de pênis é descrita como sendo associada a uma variedade de alterações morfológicas. Recentemente,houve um aumento de conhecimento acerca da patogênese do câncer de pênis, dos fatores de risco associados ao desenvolvimento das lesões precursoras relacionadas com essa doença. Embora o carcinoma de pênis seja reconhecido como um processo que ocorre em várias etapas, demonstrando um perfil policlonal, uma parte dos carcinomas de pênis é atribuída à infecção pelo HPV de alto risco, enquanto nos outros carcinomas de pênis, mecanismos moleculares independentes do HPV podem apresentar papel subjacente relevante. Entretanto, mais pesquisas sobre os mecanismos por trás da carcinogênese são necessárias.

Penile carcinoma is an uncommon and potentially mutilating disease with a still unknown etiology. Human papillomavirus (HPV) infection seems to playan important role in the development of a subset of these carcinomas and its presence is thought to be related to the histological type of the lesion. HPV prevalence in penile tumors is reported to be associated to a variety of morphological changes. In recent years, increased insight has been gained into the pathogenesis of penile cancer, the risk factors associated with penile cancer development and the clinical and histological precursor lesions related to this disease. Although penile carcinoma is recognized to be a multi-step process showing a polyclonal profile, a proportion of penile carcinoma is attributable to high risk HPV infection, while in the remaining penile cancers molecular mechanisms independent of HPV are likely to represent the more common underlying events. However, research on the mechanisms behind penile carcinogenesis is warranted.

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.

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.

Comparação de dois pares de oligonucleotídeos utilizados na reação em cadeia da polimerase para detecção de Papilomavírus Humanos em esfregaços cervicais / Comparison of two pairs of primers used in polymerase chain reaction for the detection of Hunan Papillomaviruses in cervical smears

Introdução: o câncer de colo de útero é a neoplasia mais frequente em mulheres de países em desenvolvimento. Grande parte desses casos é causada por infecção persistente com diferentes tipos de papilomavírus humano (HPV) classificados em alto e baixo risco de acordo com o potencial oncogênico. Atualmente, acredita-se que a melhor rotina de identificação e acompanhamento das lesões por HPV de forma a prevenir o câncer cervical é a combinação da técnica de Papanicolaou com a reação em cadeia por polimerase (PCR) na qual ocorre a amplificação de regiões conservadas do genoma viral, com uso de primers degenerados e em seguida identificação do tipo viral. O primer mais utilizado em todo o mundo é o MY09/11, com boa sensibilidade e especificidade. Recentemente, foi descrito na literatura um novo conjunto de primers consensuais denominado PGMY reformulando o primer MY e adicionando um novo primer HMBO visando diminuir perdas do MY (falso negativo). Objetivo: comparar os dois pares de primers, MY e PGMY, a fim de apontar aquele mais adequado para o rastreamento de infecções causadas por HPV no colo uterino pela técnica de reação em cadeia da polimerase. Métodos: avaliamos 116 amostras de esfregaços cervicais. Após a extração do DNA, a técnica de PCR foi realizada de acordo com os protocolos descritos na literatura. Resultados: através desse trabalho, observamos que o par de primers PGMY apresenta maior sensibilidadee especificidade na detecção do DNA do HPV quando comparado com o par de primers MY, além de melhores valores preditivos negativo e positivo. Conclusão: o novo par de primers PGMY, deve ser usado para substituir o par MY a fim de melhorar a detecção do DNA viral.

Introduction: cervical cancer is the most frequent neoplasia among the women of countries in development. Great part of those cases is caused by persistent infection with different types of Human Papillomavirus (HPV) classified as high and low risk, according to the risk of cervical cancer development. Nowadays, it is believed that the best identification routine and follow up of the lesions in order to prevent the malignant transformation is the combination of the technique of Papanicolaou with the polymerase chain reaction (PCR), in which the amplification of conserved areas of the viral genome occurs, with use of degenerate primers, followed by type identification. The degenerate primers MY 09/11 are used worldwide, presenting good sensibility and specificity. Recently, a new group of consensual primers denominated PGMY was described in the literature reformulating the primer MY and adding a new primer HMBO seeking to reduce losses of MY (false negative). Objective: compare two pairs of primers, MY and PGMY, to discorer the most appropriate for the diagnosis of infections caused by HPV in the uterine cervix for the technique of Polymerase chain reaction. Methods: a hundred and sixteen samples from cervical smears were evaluated. After DNA extraction, the PCR was done according to the protocols described in the literature. Results: we observed that the primers pair PGMY presents better sensibility and specificity in the detection of DNA of HPV when compared to the primers pair MY, it also presents better negative and positive predictive values. Conclusion: the new primers pair PGMY should be used to substitute the pair MY to improve the detection of the viral DNA.

Avaliação do uso combinado do teste de triagem de papanicolaou e da reação em cadeia da polimerase para identificação de pacientes em risco de câncer cervical / Evaluation of the combined use of papanicolaou screen test and polymerase chain reaction for the identification of patients at risk of cervical cancer

Introduction: in the last few years, the interest on Human Papillomavirus (HPV) has emerged due to the evidence of their carcinogenic potential,especially on the female genital tract. In the last fi fty years, the screening diagnosis has been carried out by Papanicolaou test (Pap). Nevertheless,literature describes a high rate of false-negative and false-positive samples. Objective: since an early detection is crucial to diminish the risk of cervical cancer, we aimed to analyze the use of HPV DNA detection by PCR as complementary test to routine screening. Methods: nearly 450 female smears were obtained from the DNA Bank of the Virological Diagnosis Laboratory from UFF. HPV prevalence was evaluated by using MY09/11 consensus primers and was compared to Pap test. Results: our results showed that 67.5% of the studied samples were normal tissues, among them 85.6% were negative by PCR but 14.4% were HPV infected. The remaining 32.8% were altered by Pap test. Among them, HPV DNA detection by PCR revealed a prevalence of 56.7% in ASCUS, 87.5% in LSIL and 66,6% in carcinoma. Kappa index showed a good agreement between tests (0.80). The Positive Predictive Value was considered low (58%) pointing out that important cases may be misdiagnosed as false-negatives. On the other hand, the Negative Predictive Value was considered high (90,5%) indicating that PCR should not be used as a screening test, but as a complementary one, revealing true negatives when associated to a normal result in Pap test. HPV positive samples detected by MY PCR were typed and the prevalence obtained for the different types was: 48% HPV 16, 17% HPV 33, 13% HPV 18, 18% HPV 6 and 19% were undetermined because of non tested primers or technical problems. Conclusion: analyzing the results we concluded the combination of both tests is the best diagnostic procedure, allowing a more effi cient evaluation of cancer risk and thus helping in prevention programs.

Introdução: nos últimos anos, o interesse pelo estudo dos papilomavírus humanos (HPV) aumentou, uma vez demonstrada a evidência de seu potencial oncogênico, especialmente no trato genital feminino. A partir de 1950, o rastreamento das lesões genitais foi feito pelo teste de Papanicolaou (colpocitologia oncótica). Entretanto, a literatura o atribui altas taxas de falso-positivos e negativos. Objetivo: uma vez que a detecção precoce é crucial para prevenção do câncer cervical, analisamos o uso do PCR para detecção do DNA do HPV como teste complementar à atual rotina diagnóstica. Métodos: cerca de 450 esfregaços cervicais foram obtidos do banco de amostras do Laboratório de Diagnóstico Virológico da UFF. A prevalência do HPV foi avaliadapelo uso de primers consensuais MY09/11. Resultados: nossos resultados mostraram que 67,5% das amostras eram normais e dentre eles, 85,6% eram negativas pelo PCR, mas 14,4% estavam infectadas. As 32,5% amostras restantes tinham preventivo alterado e com alta prevalência de infecções por HPV, oscilando de 56,7% em ASCUS a 87,5% em LSIL. O índice Kappa apresentou boa concordância entre os testes (0,80). O valor preditivo positivo foi baixo (58%) indicando que casos relevantes podem ser subdiagnosticados como falso-negativos. Por outro lado, o valor preditivo positivo foi elevado (90,5%) revelando que o PCR, embora não deva ser usado como método de triagem, tem importante papel complementar ao preventivo, apontando os verdadeiros negativos, quando associado à colpocitologia oncótica normal. Amostras positivas para HPV pelo PCR-MY foram tipadas e a prevalência obtida revelou 48% HPV 16, 17% HPV 33, 13% HPV 18, 18% HPV 6 e 19% de HPV indeterminados. Conclusão: nossos resultados indicam que o uso combinado de ambos os testes parece ser a conduta diagnóstica mais adequada, permitindo avaliação efi ciente do risco de câncer e assim colaborar em programas de prevenção.

Analysis of molecular biology techniques for the diagnosis of human papillomavirus infection and cervical cancer prevention

The objective of the present study was to evaluate the usefulness of molecular methodologies to access human papillomavirus genome in the genital tract. Samples from 136 women aged 17 to 52 years old obtained from the Dr. Sérgio Franco Laboratories between 2000 and 2001, were analyzed by the hybrid capture assay and amplified by PCR with generic primers MY09/MY11 and specific primers for types 16, 18, 31, 33, 35, 58. Viral genome was detected in 71.3 percent of the samples by hybrid capture and 75 percent by amplification. When cytopathology was used as a reference method for screening lesions, hybrid capture (p=0) and amplification (p=0.002) presented positive association. The 3 methods showed absolute agreement when cytopathology confirmed papillomavirus infection and high grade intraepithelial lesion. Disagreements occurred for 10 cases: seven inflammatory cases positive by PCR and negative for hybrid capture and 3 low squamous intraepithelial lesions positive for hybrid capture but negative for amplification. In conclusion, hybrid capture was shown to be sensitive and specific enough for use in clinical routines. Moreover, the evaluation of viral load values obtained by this method were shown to be related to the severity of the lesion and merit further studies to analyze the possible association with risk of progression to malignancy.

O objetivo do nosso trabalho foi avaliar o emprego de métodos moleculares para comprovar a presença dos papilomavírus humanos no trato genital. Amostras de 136 pacientes com idades entre 17 e 52 anos, coletadas nos Laboratórios Dr. Sérgio Franco entre 2000 e 2001, foram analisadas pelas técnicas de captura híbrida e amplificação pela reação em cadeia da polimerase com primers genéricos MY09/MY11 e específicos para os tipos 16, 18, 31, 33, 35, 58. O genoma viral foi detectado em 71,3 por cento dessas amostras pela captura híbrida e 75 por cento pela reação em cadeia da polimerase. Quando a citopatologia foi usada como método de referência para rastreamento das lesões, a captura (p=0) e a amplificação (p=0,002) demonstraram associação positiva. Os três testes demonstraram concordância absoluta quando a citopatologia diagnosticou processo compatível com papilomavírus ou lesão intraepitelial de alto grau. Dez casos discordantes ocorreram: sete casos de citologia inflamatório positivos na reação em cadeia da polimerase mas negativos na captura híbrida e 3 casos de lesão de baixo grau positivas na captura híbrida e negativas pela reação em cadeia da polimerase. Concluímos que a captura híbrida apresentou sensibilidade e especificidade adequadas para uso clínico. Avaliação da carga viral obtida por esta metodologia relacionou-se com a severidade da lesão e merece estudos adicionais a fim de determinar seu valor prognóstico para o câncer.

Human papillomavirus status and cervical abnormalities in women from public and private health care in Rio de Janeiro State, Brazil / Papilomavírus humano e anormalidades cervicais em mulheres do sistema de saúde privado e público no Estado do Rio de Janeiro, Brasil

This article reports the HPV status and cervical cytological abnormalities in patients attended at public and private gynecological services from Rio de Janeiro State. It also comments the performance of each HPV DNA tests used. A set of 454 women from private health clinics was tested by routine Capture Hybrid II HPV DNA assay. Among these, 58.4 percent presented HPV and nearly 90 percent of them were infected by high risk HPV types. However, this group presented few premalignant cervical lesions and no invasive cervical cancer was registered. We also studied 220 women from low income class attended at public health system. They were HPV tested by polymerase chain reaction using My09/11 primers followed by HPV typing with E6 specific primers. The overall HPV prevalence was 77.3 percent. They also showed a high percentage of high squamous intraepithelial lesion-HSIL (26.3 percent), and invasive cervical carcinoma (16.3 percent). HPV infection was found in 93.1 percent and 94.4 percent of them, respectively. The mean ages in both groups were 31.5 and 38 years, respectively. In series 1, HPV prevalence declined with age, data consistent with viral transient infection. In series 2, HPV prevalence did not decline, independent of age interval, supporting not only the idea of viral persistence into this group, but also regional epidemiological variations in the same geographic area. Significant cytological differences were seen between both groups. Normal and benign cases were the most prevalent cytological findings in series 1 while pre-malignant lesions were the most common diagnosis in the series 2. HPV prevalence in normal cases were statistically higher than those from series 1 (p < 0.001), indicating a higher exposure to HPV infection. Women from both samples were referred for previous abnormal cytology. However, socio-demographic evidence shows that women from series 1 have access to treatment more easily and faster than women from series 2 before the development of pre-malignant lesions...

Este artigo analisa a infecção por HPV e anormalidades citológicas cervicais encontradas em pacientes atendidas em serviços ginecológicos dos sistemas de saúde público e privado do estado do Rio de Janeiro. O trabalho também avalia os testes utilizados para detecção de DNA do HPV em cada população estudada. Um grupo de 454 mulheres oriundas de serviços da rede privada de saúde foi testado por Captura do Híbrido II. Destas, 58,4 por cento apresentaram infecção por HPV e cerca de 90 por cento delas estavam infectadas por HPV de alto risco. Este grupo, entretanto, apresentava poucos casos de lesões cervicais pré-malígnas e nenhum caso de câncer. Estudamos, também, 220 mulheres de baixo nível econômico atendidas no serviço de saúde pública que foram testadas para HPV pela reação da polimerase em cadeia utilizando-se os oligonucleotídeos My09/My11. A identificação dos tipos foi efetuada por amplificação com oligonucleotídeos específicos para a região E6 do genoma viral. A prevalência de HPV nesta população foi de 77.3 por cento, observando-se uma alta porcentagem de casos de neoplasias intraepiteliais cervicais de alto grau (26,3 por cento) e de carcinoma cervical invasivo (16,3 por cento). A infecção por HPV foi achada em, respectivamente, 93,1 por cento e 94,4 por cento destes casos. A média de idade em ambos os grupos era de 31,5 e 38 anos, respectivamente. Na série 1, a prevalência da infecção por HPV decresce com a idade, enquanto na série 2 ela não desaparece, dando suporte não só à idéia de persistência viral neste grupo, mas também a variações epidemiológicas na mesma área geográfica. Diferenças significativas foram vistas nos dois grupos. Casos normais e benignos foram incidentes na série 1, enquanto as lesões malígnas predominaram na série 2. Ao contrário, casos normais infectados por HPV eram prevalentes na série 2 (p < 0.001), indicando maior exposição ao vírus. Embora as mulheres de ambos os grupos tenham sido incluídas no estudo por apresentarem citologia anormal, evidências sócio-demográficas demonstram que mulheres da série 1 tem acesso mais fácil e rápido ao tratamento do que as mulheres da série 2 antes que as lesões pré-malígnas se desenvolvam...

Prevalence of human papillomavirus infection in the genital tract determined by hybrid capture assay

Human Papillomavirus (HPV) infection is the most prevalent sexually-transmitted virus worldwide. It is known to be the etiological agent of cervical cancer and cervical intraepithelial neoplasia (CIN). Consequently, there is strong motivation to evaluate HPV testing in cervical cancer screening. Recently developed, the second generation of the hybrid capture test (HCA II) is a non-radioactive, relatively rapid, hybridization assay, designed to detect 18 HPV types divided into high and low-risk groups. We evaluated 7,314 patients (5,833 women and 1,481 men) for HPV infection by HCA II. Among them, 3,008 (41.1 percent) presented HPV infection: 430 (14.2 percent) had HPV DNA of low risk for cancer, 1,631 (54.2 percent) had high risk HPV types and 947 (31.5 percent) had both types. The prevalence in females was 44.9 percent. The prevalence of HPV DNA in the group for which cytological results were available was slightly higher: 55.3 percent (1007/1824). Significant differences were detected in the frequency of HPV infection of the cervix between normal cases and those with high-grade squamous-intraepithelial lesions (HSIL)(P<0.0001). Among males, the prevalence was 26.2 percent, composed of 9.1 percent in Group A, 9.7 percent in Group B and 7.4 percent with multiple infections. We observed that male prevalence was lower and that low-risk types were more frequent than in females. HPV viral load was significantly greater in SILs than in normal or inflammatory cases (P<0.0001), suggesting an association between high viral load values and risk of SIL. Because of high costs, the HCA II test cannot be recommended for routine mass screening for cervical infection in poor countries. Nevertheless, it was found to be a useful tool, when combined with cytology, discovering high-risk infections in apparently normal tissues and revealing silent infections that may be responsible for the maintenance of HPV in the general population. These findings point...

Detecção de papilomavírus humanos pela técnica de captura do híbrido no trato genital masculino / Human papillomavirus detection by the hybrid capture assay in the male genital tract

Estudo das infecções genitais causadas pelo HPV, determinando a prevalência dos tipos de HPV noa pacientes masculinos

Comparative analysis of the polymerase chain reaction and the hybrid capture assay for the detection of human papillomavirus infection

Avalia o emprego de métodos moleculares a fim de comprovar a presença dos HPV no trato genital para utilização em combinação com a citopatologia, método utilizado para rastreamento das lesões causadas pelo HPV

Detection of human papillomavirus DNA by the hybrid capture assay

Human Papillomavirus (HPV) infection is the main cause of cervical cancers and cervical intraepithelial neoplasias (CIN) worldwide. Consequently, it would be useful to evaluate HPV testing to screen for cervical cancer. Recently developed, the second-generation Hybrid Capture (HCA II) test is a non-radioactive, relatively rapid, liquid hybridization assay designed to detect 18 HPV types, divided into high and low-risk groups. We evaluated 1055 women for HPV infection with the HCA II test. Five hundred and ten (48.3 percent) of these women had HPV infection; 60 (11.8 percent) had low cancer-risk HPV DNA; 269 (52.7 percent) had high-risk HPV types and 181 (35.5 percent) had both groups. Hence, 450 women (88.2 percent) in this HPV-infected group had at least one high risk HPV type, and were therefore considered to be at high risk for cancer. Among the group with Papanicolaou (Pap) test results, the overall prevalence of HPV DNA was 58.4 percent. Significant differences in HPV infection of the cervix were detected between Pap I (normal smears) and Pap IV (carcinomas) (p<0.0001). Values of HPV viral load obtained for Pap I and SILs were significantly different, with an upward trend (p<0.0001), suggesting a positive correlation between high viral load values and risk of SIL. Because of the high costs of the HCA II test, its use for routine cervical mass screening cannot be recommended in poor countries. Nevertheless, it is a useful tool when combined with cytology, diagnosing high-risk infections in apparently normal tissues. Use of this technique could help reduce the risk of cancer

HPV 16 detection in cervical lesions, physical state of viral DNA and changes in p53 gene

CONTEXT: Persistent infection with high risk human papillomavirus (HPV) has been linked to cervical carcinoma. Integration of viral DNA into host cell DNA is essential for this cancer development, promoting disruption of the HPV E2 gene, thus leading to unregulated increases in E6 and E7 proteins and inactivating the products of p53 and Rb tumor suppressor genes. OBJECTIVE: To investigate HPV 16 infection in cervical lesions, physical state of viral DNA and p53 gene alterations in a group of women attending a public health service. DESIGN: Prospective, non-controlled, transversal study. SETTING: Gynecological clinic of the School od Medicine, Universidade Federal Fluminense. SAMPLE: 43 consective patients with cervical lesions referred to our service. MAIN MEASUREMENTS: Cases were classified via cytology/histology as normal, HPV infection, condyloma, low-grade squamous intraepithelial lesion (LSIL), high-grade squamous intraepithelial lesion (HSIL) and carcinoma. HPV infection was studied via polymerase chain reaction (PCR) using two PCR primer sets, to determine DNA integration. p53 gene changes were investigated by single-strand conformation polymorphism (SSCP) analysis. RESULTS: One normal case, 7 HPV infections, 6 condylomas, 7 LSIL, 14 HSIL and 8 cancers were found, with 95 percent positive for HPV genome when tested using both L1 and E6 primers. HPV 16 was most prevalent (73.1 percent). HPV 16 DNA was integrated within the host genome in 3 LSIL. One LSIL progressed to HSIL by 13 months after first diagnosis. Among HPV 16-positive HSIL cases, 50 percent contained integrated viral DNA. HPV 16 E2 gene disruption was seen in 7 cancers (87.5 percent). Only smal-cell carcinoma showed intact HPV 16 E2 gene. Abnormal p53 bands detected by PCR/SSCP were observed in 4 cases: 2 squamous carcinoma with parametrium (exon 8) and two cervical intraepithelial neoplasia (CIN) III (exons 5 and 7). All cases presented HPV 16 E2 gene loss. CONCLUSIONS: The sample had a high rate of high-risk HPV detected in benign and malignant lesions; high cervical cancer burden; HPV 16 DNA integration in all except one case of cancer; p53 gene changes in CIN III and in invasive cancer cases associated with DNA integration

Analysis of the p53 gene and papillomavirus detection in smears from cervical lesions

CONTEXT: Alterations of the p53 tumor suppressor gene are correlated with a critical step in the development of many human cancers.The tumor suppressor gene functions include regulation of the cell cycle and the cellular response to DNA damage, initiation of DNA repair and replication, induction of apoptosis and promotion of cell differentiation. CASE REPORT: Smears from ten cases of cervical lesions were analyzed for status of exons 5-8 of the p53 gene using PCR/SSCP. HPV infection was also screened by the PCR method using two PCR primer sets. Changes in the p53 gene were observed in a case of squamous carcinoma and a case of asymptomatic cervical intraepithelial neoplasia grade III (CIN III). High-risk HPV was detected in both cases showing that HPV infection and p53 mutation are not exclusive events

Condilomas acuminados extragenital / Extragenital warts

Descrevemos dois casos clínicos de condilomas acuminados extragenital, ambos em cicatriz de cirurgia abdominal. Um caso, foi em paciente do sexo feminino que mantinha relaçäo sexual com pessoa do mesmo sexo. Ela tinha, concomitante às lesöes em parede abdominal, também lesöes de condiloma acuminado em vulva e NIC 1. O segundo, ocorreu em pacientes do sexo masculino que näo tinha lesäo por HPV em genital. Ambos relataram que seus parceiros sexuais eram únicos e que tinham tido lesöes genitais verrucosas. Seräo discutidos os possíveis motivos que propiciaram as infecçöes