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.

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.

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.

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.

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.

Human Papillomavirus infection in men residing in Brazil, Mexico, and the USA / Infección por Virus de Papiloma Humano en hombres de Brasil, México y EUA

OBJECTIVE: To assess Human Papillomavirus (HPV) type distribution among men ages 18 years and older recruited from three different countries utilizing a common protocol for sampling HPV detection, and to evaluate whether HPV detection differs by age and country. MATERIAL AD METHODS: The study protocol includes a pre-enrollment run-in visit, a baseline (enrollment) visit, and nine additional visits after enrollment scheduled six months apart. For this analysis, the first 1160 men who completed both the run-in and baseline visit were included. To maximize sampling and prevent fraying of applicators, three different applicators were utilized to sample the external genitalia of participants among different anatomic sites. These samples were later combined to form a single sample for the detection of HPV using polymerase chain reaction (PCR) for amplification of a fragment of the HPV L1 gene. RESULTS: Among 1160 men from Brazil, Mexico, and the United States (U.S.), overall HPV prevalence was 65.2 percent; with 12.0 percent oncogenic types only, 20.7 percent non-oncogenic types only, 17.8 percent both oncogenic and non-oncogenic, and 14.7 percent unclassified infections. Multiple HPV types were detected in 25.7 percent of study participants. HPV prevalence was higher in Brazil (72.3 percent) than in the U.S. (61.3 percent) and Mexico (61.9 percent). HPV 16 (6.5 percent), 51 (6.5 percent), and 59 (5.3 percent) were the most commonly detected oncogenic infections, and HPV 84 (7.7 percent), 62 (7.3 percent), and 6 (6.6 percent) were the most commonly detected non-oncogenic infections. Overall HPV prevalence was not associated with age. However, significant associations with age were observed when specific categories of oncogenic, non-oncogenic, and unclassified HPV infections were considered. CONCLUSIONS: Studies of HPV type distribution among a broad age range of men from multiple countries is needed to fill the information gap internationally ...

OBJETIVO: Evaluar la distribución tipo específica de infección por Virus del Papiloma Humano (VPH) entre hombres de 18 años o más de tres países con un protocolo común para el muestreo de la detección de VPH, y evaluar si la detección de VPH varía de acuerdo con la edad y el país. MATERIAL Y MÉTODOS: El estudio incluye diversas etapas que inician con la identificación de hombres susceptibles, una medición basal (visita de enrolamiento) y nueve visitas adicionales programadas cada seis meses. En este artículo, se presenta el análisis de los primeros 1160 hombres que fueron incluídos en el estudio. Para maximizar la posibilidad de detección de VPH se utilizó un cepillo de dacrón que muestreó en forma combinada diferentes sitios anatómicos. Para la determinación de ADN de VPH se utilizó ión en cadena de polimerasa (PCR) por amplificación de un fragmento del gen de VPH L1. RESULTADOS: Entre 1160 hombres de Brasil, México y EUA, la prevalencia global de VPH fue de 65.2 por ciento, con solamente 12 por ciento de tipos oncogénicos, 20.7 por ciento de tipos de VPH no oncogénicos, 17.8 por ciento de muestras positivas a tipos oncogénicos y no oncogénicos; y finalmente 14.7 por ciento de infecciones no clasificadas. Múltiples tipos de VPH fueron detectados en 25.7 por ciento de los participantes en el estudio. La prevalencia de VPH fue más alta en Brasil (72.3 por ciento), comparada con la observada en EUA (61.3 por ciento) y México (61.9 por ciento). Los tipos de VPH 16 (6.5 por ciento), 51 (6.5 por ciento) y 59 (5.3 por ciento) fueron los más comúnmente observados con poder oncogénico. El VPH 84 (7.7 por ciento), 62 (7.3 por ciento) y 6 (6.6 por ciento) fueron las infecciones no oncogénicas más comunes. CONCLUSIONES: Son necesarios estudios de la distribución de VPH en un amplio margen de edad entre hombres de múltiples países, para establecer con mayor precisión, el conocimiento de la historia natural de la infección por VPH en hombres.

Prevalência do papilomavírus humano (HPV) na cavidade oral e na orofaringe / Prevalence of human papillomavirus (HPV) in oral cavity and oropharynx

A prevalência do papilomavírus humano (HPV) na cavidade oral e na orofaringe ainda não está bem esclarecida como nos estudos do trato genital, na qual é bem definida. Entretanto, novas pesquisas estão surgindo após o aparecimento dos exames de biologia molecular. Neste estudo foi realizada uma revisão da literatura com o objetivo de verificar a prevalência do papilomavírus humano na cavidade oral e na orofaringe. Os resultados desta pesquisa mostraram uma prevalência do HPV 16 na mucosa oral normal (infecção latente). Já nas lesões benignas orais associadas ao HPV mostraram uma prevalência do HPV 6 e 11 em papilomas de células escamosas e condilomas, e, nas verrugas, uma prevalência do HPV 2 e 57, enquanto na hiperplasia epitelial focal prevaleceram os HPVs 13 e 32, e no câncer oral, principalmente, no carcinoma de células escamosas (CCE), foi evidenciada uma alta prevalência do HPV 16, o que sugere sua participação na carcinogênese oral, apesar de ser um assunto controverso. Constatou-se também uma enorme discrepância nos resultados da prevalência do papilomavírus humano (HPV) na mucosa oral normal (infecção latente) e no câncer oral, enquanto nas lesões benignas associadas ao vírus, os resultados foram confirmatórios.