Análisis del uso de las técnicas de tamizaje para la detección de cáncer cervicouterino en Chile / Analysis of the use of screening techniques for the detection of cervical cancer in Chile

El Cáncer Cervicouterino es una de las principales problemáticas en salud de las mujeres a nivel mundial. Las técnicas de tamizaje para la detección de Cáncer Cervicouterino han ido desarrollándose a través de la historia reciente tanto intencionada como casualmente, incorporando con cada nuevo descubrimiento a las nuevas tecnologías. Actualmente existen diversos procedimientos vigentes que van desde pruebas sencillas de inspección visual, pasando por las tradicionales muestras citológicas, hasta la utilización de tecnología molecular, y los usos que se les dan a estos métodos de cribado potencian la detección temprana del Cáncer Cervicouterino, razón por lo cual se realiza una revisión respecto de las técnicas de tamizajes disponibles y su aplicación en Chile.

Cervical Cancer is one of the main problems in women's health worldwide. Screening techniques for the detection of Cervical Cancer have been developed throughout recent history both intentionally and incidentally, incorporating new technologies with each new discovery. Currently there are various procedures in force that range from simple visual inspection tests, through traditional cytological samples, to the use of molecular technology, and the uses that are given to these screening methods enhance the early detection of Cervical Cancer, which is why which is carried out a review regarding the available screening techniques and their application in Chile.

Virus del papiloma humano en carcinoma colorrectal / Human papilloma virus in colorectal carcinoma

Resumen Introducción: Previos trabajos han reportado una asociación entre la infección del virus del papiloma humano (VPH) y el desarrollo de cáncer colorrectal, aunque existe controversia al respecto. Materiales y Método: Estudio observacional, transversal, descriptivo, retrospectivo, no ciego. Se utilizaron 50 muestras de patología con diagnóstico de adenocarcinoma colorrectal, incluidas en parafina, para aislar ADN de las muestras. Se realizó la extracción de ADN mediante protocolos establecidos para extracción, lisis y rehidratación de muestra. Se identificó y genotipicó el ADN del virus para amplificar y detectar subtipos oncogénicos de entre 35 subtipos diferentes incluidos en la prueba, secuenciando las muestras positivas, utilizando protocolos ya establecidos de purificación y análisis de muestra, mediante microarreglos. Resultados: Se identificaron 14 muestras de 50 (28%) estudiadas positivas para el virus de papiloma humano de las cuales 11 (22%) incluyen uno o más subtipos de alto riesgo para neoplasia. No se identificaron diferencias estadísticamente significativas entre grupos en cuanto a edad, sexo, localización del tumor, grado de diferenciación, infiltración, ganglios afectados, metástasis o número de paquetes/año. Conclusión: La detección de los subtipos de VPH de alto riesgo en un alto porcentaje de las muestras positivas, sugiere una asociación entre la infección con el desarrollo de cáncer colorrectal.

Introduction: Previous works have reported an association between human papilloma virus (HPV) infection and the development of colorectal cancer, and although controversy regarding this association exists. Materials and Method: This was an observational, cross-sectional, descriptive, retrospective unblinded study. Fifty pathology samples embedded in paraffin with a diagnosis of colorectal adenocarcinoma were used to isolate DNA from the tissue. DNA was extracted according to established protocols for extraction, lysis and sample rehydration. Viral DNA was identified and genotypified to amplify and detect oncogenic subtypes among 35 different subtypes included in the study, sequencing positive samples with established protocols of purification and sample analysis using microarrays. Results: Fourteen of 50 (28%) samples were identified as positive for human papilloma virus; of these 11 (22%) included one or more high-risk subtypes for neoplasia. Statistically significant differences were not found between the groups regarding age, sex, tumor location, degree of differentiation, infiltration, affected lymph nodes, metastasis and number of pack years. Conclusion: The detection of high-risk VPH subtypes in a high percentage of positive samples, suggests an association between infection and the development of colorectal cancer.

Optimización de unidades formadoras de colonias y detección del virus de papiloma humano en sangre de cordón umbilical / Colony-forming units optimization and human papillomavirus detection in umbilical cord blood

Resumen Introducción: Se requiere analizar diversos parámetros para el control de calidad adecuado de las unidades de sangre de cordón umbilical (USCU) cuando se utilizan con fines terapéuticos. Objetivo: Optimizar las unidades formadoras de colonias (UFC) de cultivos clonogénicos y detectar el genoma del virus del papiloma humano (VPH) en USCU. Métodos: Se incluyeron 141 muestras de sangre de cordón umbilical (SCU), de segmento y de UFC de cultivos clonogénicos de USCU. Se realizó extracción de ADN, cuantificación y amplificación por PCR del gen endógeno GAPDH. Se detectó el gen L1 del VPH con los oligonucleótidos MY09/MY11 y GP5/GP6+; los productos de PCR se migraron en electroforesis de agarosa. El ADN purificado de las UFC se analizó mediante electroforesis de agarosa y algunos ADN, con la técnica sequence specific priming. Resultados: La concentración de ADN extraído de UFC fue superior comparada con la de SCU (p = 0.0041) y la de segmento (p < 0.0001); así como la de SCU comparada con la de segmento (p < 0.0001). Todas las muestras fueron positivas para la amplificación de GAPDH y negativas para MY09/MY11 y GP5/GP6+. Conclusiones: Las USCU criopreservadas fueron VPH netativas; además, es factible obtener ADN en altas concentraciones y con alta pureza a partir de UFC de los cultivos clonogénicos.

Abstract Introduction: Analysis of several markers is required for adequate quality control in umbilical cord blood units (UCBU) when are used for therapeutic purposes. Objective: To optimize colony-forming units (CFU) from clonogenic cultures and to detect the human papillomavirus (HPV) genome in UCBU. Methods: One hundred and forty-one umbilical cord blood (UCB), segment or CFU samples from UCBU clonogenic cultures were included. DNA extraction, quantification and endogenous GAPDH gene PCR amplification were carried out. Subsequently, HPV L1 gene was detected using the MY09/MY11 and GP5/GP6+ oligonucleotides. PCR products were analyzed with electrophoresis in agarose gel. CFU-extracted purified DNA was analyzed by electrophoresis in agarose gel, as well as some DNAs, using the SSP technique. Results: CFU-extracted DNA concentration was higher in comparison with that of UCB (p = 0.0041) and that of the segment (p < 0.0001), as well as that of UCB in comparison with that of the segment (p < 0.0001). All samples were positive for GAPDH amplification and negative for MY09/MY11 and GP5/GP6+. Conclusions: Cryopreserved UCBUs were HPV-negative. Obtaining CFU DNA from clonogenic cultures with high concentrations and purity is feasible.

Características operativas de técnicas moleculares empleadas para la detección del virus del papiloma humano en el proyecto ESTAMPA / Operational characteristics of molecular techniques used to detect human papillomavirus in the ESTAMPA project

Resumen El desarrollo y la innovación de nuevas tecnologías ha permitido mejorar la detección de la infección por el virus del papiloma humano de alto riesgo. La captura de híbridos II es un ensayo que se basa en hibridación y quimioluminiscencia. Cobas VPH Test es una PCR cualitativa y Aptima VPH Assay permite detectar la expresión de ARN mensajero de las oncoproteínas E6/E7 del VPH de alto riesgo. Estas técnicas presentan ventajas en comparación con la citología convencional, que se utiliza como prueba de rutina para la detección temprana del cáncer de cuello uterino. En el estudio ESTAMPA se realizaron 13.691 procesamientos que permitieron identificar que para el planteamiento de proyectos de investigación o para la implementación de pruebas de tamizaje de VPH es necesario analizar las ventajas y desventajas de las pruebas del mercado.

Abstract The development and innovation of new technologies has improved the detection of high-risk human papillomavirus infection. Hybrid capture II is an assay that is based on hybridization and chemiluminescence. Cobas HPV Test is a qualitative PCR and Aptima HPV Assay allows to detect the expression of messenger RNA of the high- risk HPV E6 / E7 oncoproteins. These techniques have advantages, in comparison, with conventional cytology that is routinely used for the detection of cervical cancer. In the ESTAMPA study, 13,691 prosecutions were carried out that allowed to identify that for the planning of research projects or for the implementation of HPV screening tests, it is necessary to analyze the advantages and disadvantages of market tests.

High-risk human papillomavirus testing for triage of women with previous cytological abnormalities from the Vale do Ribeira region / Teste de papilomavírus humano para triagem de mulheres com alterações em citologia anterior provenientes da região do Vale do Ribeira

Abstract Objective To evaluate the performance of the hybrid capture 2 (HC2) high-risk papillomavirus (hrHPV) assay and cytological test in women with previous abnormalities, to detect cervical intraepithelial neoplasia grade 2 or worse (≥ CIN 2). Methods A cytological test and HC2 (Qiagen, Gaithersburg, Maryland, EUA) for hrHPV were conducted in 359 liquid-based (Sure Path, Becton Dickinson, TriPath Imaging, Burlington, NC, USA) samples collected from women from the Vale do Ribeira Region, during July 2013 and September 2015 with previous cytology classified as atypical squamous cells of undetermined significance (ASC-US), low-grade squamous intraepithelial lesion (LSIL), atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesions (ASC-H), and atypical glandular cells (AGC). The histopathological examination was conducted in 179 women. The performance evaluations were calculated using the "exact" Clopper-Pearson 95% confidence interval (CI) test by MEDCALC (Medcalc Software Ltd, Ostend, Belgium). Results The ≥ CIN 2 frequency was 11.7% (21/179). The HC2 for hrHPV and repeat cytology to detect ≥ CIN 2 obtained, respectively, a sensitivity of 90.5% (95% CI = 69.6-98.8) and 90.5%, (95%CI = 69.6-98.8), a specificity of 65.8% (95% CI = 57.9-73.2) and 43.7% (95%CI = 35.8-51.8), a positive predictive value of 26.0% (95% CI = 21.4-31.3) and 17.6%, (95%CI = 14.9-20.6), and a negative predictive value of 98.1% (95%CI = 93.3-99.5) and 97.2% (95% CI = 90.1-99.2). Conclusion Hybrid capture 2 for hrHPV improves the performance of the detection of ≥ CIN 2, without compromising sensitivity, and provides a greater safety margin to return to the triennial screening of women undergoing follow-up due to previous abnormalities, without underlying ≥ CIN 2.

Resumo Objetivo Avaliar o desempenho da captura híbrida 2 (CH2) para papilomavírus humano de alto risco (HPVar) e repetição do exame citopatológico em mulheres com anormalidades em citologia anterior, para detectar neoplasia intraepitelial cervical grau 2 ou pior (≥ NIC 2). Métodos Foi realizado exame citopatológico e CH2 para HPVar (Qiagen, Gaithersburg, Maryland, EUA) em 359 amostras em meio líquido (Sure Path, Becton Dickinson, TriPath Imaging, Burlington, NC, USA) coletadas de mulheres da região do Vale do Ribeira, durante julho de 2013 e setembro de 2015 com citologia anterior classificada como células escamosas atípicas de significado indeterminado (ASC-US), lesão intraepitelial de baixo grau (LSIL), células escamosas atípicas, não podendo excluir lesão de alto grau (ASC-H) e células glandulares atípicas (AGC). O exame histopatológico foi realizado em 179 mulheres. As avaliações de desempenho foram calculadas usando o teste de intervalo de confiança (IC) "exato" de Clopper-Pearson de 95% pelo software MEDCALC (Medcalc Software Ltd, Ostend, Bélgica). Resultados A frequência de≥ NIC 2 foi 11,7% (21/179). A CH2 para o HPVar e a citologia de repetição para a detecção ≥ NIC 2 obteve, respectivamente, sensibilidade de 90.5% (IC 95% = 69,6-98,8) e 90,5% (IC 95% = 69,6-98,8), especificidade de 65,8% (IC 95% = 57,9-73,2) e 43,7%, (IC 95% = 35,8-51,8), valor preditivo positivo de 26,0% (IC 95% = 21,4-31,3) e 17,6%, (IC95% = 14,9-20,6), e valor preditivo negativo de 98,1% (IC 95% = 93,3-99,5) e 97,2%, (IC 95% = 90,1-99,2). Conclusão No geral, a CH2 para HPVar aprimora o desempenho para detecção de ≥ NIC 2, sem comprometer a sensibilidade e proporciona maior margem de segurança do retorno ao rastreio trienal de mulheres com anormalidades citológicas prévias, sem≥ NIC 2 subjacente.

Avaliação do perfil de mulheres atendidas em centros de referência em saúde de Porto Alegre/RS e relação de alterações citológicas detectadas no exame citopatológico e a presença do HPV / Profile evaluation of women that attended reference health centers in Porto Alegre/RS and the relationship between cytological alterations detected in cytopathological examination and presence of HPV / Evaluación del perfil de las mujeres atendidas en los centros de referencia de salud de Porto Alegre / RS y la relación de las alteraciones citológicas detectadas en el examen citopatológico con la presencia de VPH

Justificativas e Objetivos: Câncer de colo de útero é considerado um problema de saúde pública mundial. Seu diagnóstico é realizado através do exame citopatológico (EC) e seu desenvolvimento relacionado à infecção pelo papilomavírus humano (HPV). Este estudo objetiva avaliar o perfil de mulheres atendidas em centros de referência em saúde de Porto Alegre, Rio Grande do Sul, assim como a relação de alterações observadas ao EC com presença do HPV. Métodos: Estudo transversal realizado em mulheres atendidas em unidades básicas de saúde e um ambulatório de referência de hospital público terciário, no período de julho de 2014 a janeiro de 2017. Coletaram-se amostras representativas da endo/ectocérvice para realização do EC e investigadas quanto à presença molecular do HPV. Resultados: Foram analisadas 169 mulheres com idade média entre 31 e 40 anos, das quais 125 (74%) informaram que a sexarca ocorreu na faixa de 15-20 anos e 37,9% relatou ter tido de três a cinco parceiros sexuais. Em relação ao EC, 71 (42%) apresentaram resultado negativo para lesão intraepitelial ou malignidade e 98 (58%) alguma anormalidade de células escamosas: 20 (11,8%) atipias; 22 (13%) lesão intraepitelial escamosa de baixo grau e 56 (32,6%) lesão intraepitelial de alto grau (HSIL). Cinquenta (29,6%) apresentaram positividade para HPV, destas 56,4% foram diagnosticadas com HSIL (p<0,01). Conclusão: Os resultados revelam alta frequência de HPV em amostras com alterações citopatológicas, em mulheres jovens e com grau de exposição ao HPV, reforçando a importância do papel da sua identificação precoce na investigação da carcinogênese cervical.(AU)

Background and Objectives: Cervical cancer is considered a worldwide public health problem. Its diagnosis is made through cytopathological examination and its development related to human papillomavirus (HPV) infection. This study aims to evaluate the profile of women treated at reference health centers in Porto Alegre, Rio Grande do Sul, as well as the relation of changes observed to cytopathological examination with the presence of HPV. Methods: This is a cross-sectional study carried out in women treated at basic health units and a referral clinic of a public tertiary hospital, from July 2014 to January 2017. Representative samples of the endo/ectocervix were collected to perform the cytopathological examination and investigated for the molecular presence of HPV. Results: 169 women with mean age between 31 and 40 years were analyzed, of whom 125 (74%) reported that the onset of sexual activity occurred in the 15-20 years age group, and 37.9% reported having had three to five sexual partners. In relation to cytopathological examination, 71 (42%) had a negative result for intraepithelial lesion or malignancy and 98 (58%) some squamous cell abnormality: 20 (11.8%) atypical; 22 (13%) low-grade squamous intraepithelial lesion and 56 (32.6%) high-grade intraepithelial lesion (HSIL). Fifty (29.6%) were positive for HPV, of which 56.4% were diagnosed with HSIL (p < 0.01). Conclusion: The results reveal a high frequency of HPV in samples with cytopathological changes, in young women and with a degree of exposure to HPV, reinforcing the importance of the role of its early identification in the investigation of cervical carcinogenesis.(AU)

Justificación y objetivos: El cáncer de cuello uterino se considera un problema de salud pública en todo el mundo. Su diagnóstico se realiza mediante el examen citopatológico (EC), y su desarrollo está relacionado con la infección por el virus del papiloma humano (VPH). Este estudio objetivó evaluar el perfil de mujeres atendidas en los centros de referencia en salud de Porto Alegre, Rio Grande do Sul (Brasil), así como la relación de las alteraciones observadas en el EC con la presencia del VPH. Métodos: Estudio transversal realizado en las mujeres atendidas por unidades de atención primaria y por una clínica ambulatoria de referencia del hospital público terciario en la ciudad de Porto Alegre, en el período de julio de 2014 a enero de 2017. Se recolectaron muestras representativas de endo/ectocérvice para realizar la CE, las cuales se clasificaron según el sistema Bethesda y se investigaron la presencia molecular del VPH. Resultados: Analizamos 169 mujeres con promedio de edad entre 31 y 40 años, de las cuales 125 (74%) informaron que el sexarche ocurrió en el rango de 15-20 años. La mayoría (37,9%) informó haber tenido de 3 a 5 parejas sexuales; y el 37,3% estaban usando anticonceptivos orales. Con respecto a la EC, 71 (42%) se clasificaron como negativos para lesión intraepitelial o malignidad; y el 98 (58%) tenían alguna anormalidad de células escamosas: 20 (11,8%) de atipias; 22 (13,0%) lesión intraepitelial escamosa de bajo grado y 56 (32,6%) lesión intraepitelial de alto grado (HSIL). La frecuencia de positividad del VPH encontrada fue de 50 (29,6%), de estas un 56,4% fueron diagnosticadas con HSIL (p<0,01). Conclusiones: Estos resultados revelan una alta frecuencia de VPH en muestras con alteraciones citopatológicas presentes en mujeres jóvenes con cierto grado de exposición al VPH, lo que refuerza la importancia de identificarse tempranamente en el análisis de la carcinogénesis cervical.(AU)

Clonación y expresión en Escherichia coli de un gen L1 completo del virus del papiloma humano 18 aislado de una paciente cubana y variantes delecionadas / Cloning and expression in Escherichia coli of the full-length and deletion variants of a human papillomavirus 18 L1 gene isolated from a Cuban patients

Introduction: Human papillomavirus (HPV) vaccines are based on the L1 major capsid protein. Objectives: To clone the HPV-18 L1 gene from a Cuban female HPV-18-infected patient and to express the full-length and deletion variants of the cloned HPV-18 L1 gene in Escherichia coli. Methods: The full-length HPV-18 L1 gene was PCR-amplified from total DNA isolated from a Cuban patient, cloned and finally subcloned into the E. coli expression vector pET26b. Three deletion mutants were constructed, which encode truncated proteins lacking 30 amino acids at the C-terminus in combination with 5, 6 or none deleted residue at the N-terminus. Production of L1 proteins in E. coli BL21(DE3) and E. coli SHuffle T7 was assessed by SDS-PAGE and Western blotting. Results: The cloned HPV-18 L1 gene was 99.9 por ciento similar to the African variant EF202152 and probably shares a common origin with the B lineage of genotype 18. The three truncated variants of HPV-18 L1 were produced at higher levels than the full-length HPV-18 L1 protein, attaining higher levels in E. coli BL21(DE3) and higher solubility in E. coli SHuffle. The C-terminus-only truncated variant, L1∆C30, was produced at similar levels to the HPV-18 L1s truncated at both termini. E. coli SHuffle produced about three times more amounts of L1∆C30 when grown under autoinduction conditions with respect to conventional induction and thus, amounts were comparable to those obtained in E. coli BL21(DE3) under conventional induction. Conclusions: Truncation of thirty amino acid residues at the carboxy-terminus of the HPV-18 L1 made a major contribution to the production and solubility of this wild-type protein in E. coli. This is the first report about soluble production of HPV-18 L1 protein in an E. coli SHuffle strain. However, higher amounts of L1 are needed to scale-up its production for developing an HPV vaccine candidate(AU)

Introducción: Las vacunas contra el virus del papiloma humano (VPH) se fundamentan en la proteína principal de la cápsida L1. Objetivo: Clonar el gen L1 del VPH-18 a partir de una paciente cubana infectada con VPH-18 y expresar las variantes de longitud completa y delecionadas del gen L1 del VPH-18 en Escherichia coli. Métodos: El gen L1 del VPH-18 de longitud completa se amplificó por PCR a partir de ADN total aislado de un paciente cubana, se clonó y finalmente se subclonó en el vector de expresión de E. coli pET26b. Se construyeron tres mutantes de deleción, que codifican para proteínas truncadas que carecen de 30 aminoácidos por el extremo carboxilo, en combinación con 5, 6 o ningún residuo delecionado por el extremo amino. La producción de las proteínas L1 en E. coli BL21(DE3) y E. coli SHuffle T7 se evaluó mediante SDS-PAGE y Western blot. Resultados: El gen L1 del VPH-18 clonado fue 99.9 percent similar a la variante africana EF202152 y probablemente comparte un origen común con el linaje B del genotipo 18. Las tres variantes truncadas de la proteína L1 del VPH-18 se produjeron a mayores niveles que la proteína L1 del VPH-18 de longitud completa, alcanzando mayores niveles en E. coli BL21(DE3) y mayor solubilidad en E. coli SHuffle. La variante truncada solo por el extremo carboxilo, L1(C30, se produjo a niveles similares a las proteínas L1 del VPH-18 truncadas por ambos extremos. E. coli SHuffle produjo aproximadamente tres veces más cantidades de L1(C30 cuando creció en condiciones de autoinducción, con respecto a la inducción convencional y, por ende, las cantidades fueron comparables a las obtenidas por E. coli BL21(DE3) bajo inducción convencional. Conclusiones: La truncación de treinta residuos de aminoácidos por el extremo carboxilo de la proteína L1 del VPH-18 tuvo una importante contribución a la producción y solubilidad de la proteína L1 nativa en E. coli. Este es el primer informe sobre la producción soluble de la proteína L1 del VPH-18 en una cepa de E. coli SHuffle. Sin embargo, se necesitan mayores cantidades de la proteína L1 para escalar su producción para desarrollar un candidato vacunal contra el VPH(AU)

Frecuencia de alteraciones citológicas cervicales y virus papiloma humano en una muestra de estudiantes universitarias en Temuco, Chile / Frequency of cervical cytological alterations and human papilloma virus in a sample of university students in Temuco, Chile

Resumen Introducción: El inicio precoz de actividad sexual puede favorecer el desarrollo de alteraciones cervicales y de infecciones de transmisión sexual, en especial del virus papiloma humano (VPH) muy frecuente en adolescentes y jóvenes. Objetivo: Analizar el estado del cuello uterino, presencia del VPH y conductas sexuales en mujeres menores de 25 años. Material y Métodos: Participaron 182 estudiantes universitarias de 18-24 años, sanas, sexualmente activas y no vacunadas para VPH. Se realizó Papanicolaou (Pap) y clasificación del VPH en alto y bajo riesgo (AR y BR) mediante reacción de polimerasa en cadena en tiempo real. Las conductas sexuales fueron consultadas privadamente. Resultados: El 46,9% de los Pap presentaron alteraciones citológicas (inflamación inespecífica/hemorrágico: 29,4% y frotis atípicos (FA):10,2%). La frecuencia de los VPH-AR fue 24,3%; de éstos, 67,4% presentó un Pap alterado. Hubo asociación entre alteraciones citológicas y presencia de VPH (p < 0,0001) y años de actividad sexual y FA o neoplasia intraepitelial grado I (NIE I) (p = 0,009). El 11,9% de las jóvenes estudiadas (21/177) presentó FA o NIE I, con 66,7% de casos VPH-AR. Conclusiones: Estos hallazgos alertan la vulnerabilidad de estas jóvenes que tendrían un riesgo potencial de persistencia viral, NIE y eventualmente cáncer. Es importante enfatizar consejería y prevención previo a la edad normada de ingreso al programa de cribado para cáncer cérvico uterino en Chile.

Background: The early onset of sexual activity can promote the development of cervical alterations and sexually transmitted infections, especially the human papillomavirus (HPV) very common in adolescents and young people. Aim: The condition of the cervix, HPV and sexual behavior in young women under 25 years of age were analyzed. Methods: 182 university students, healthy, sexually active, 18-24 years old, without vaccine for HPV participated. Papanicolaou (Pap) test and classification of high and low risk HPV (HR and LR) were performed by real time polymerase chain reaction. The sexual behaviors were consulted in private. Results: The 46.9% of Pap presented cytological alterations, non-specific inflammation/hemorrhagic (29.4%) and atypical smear (10.2%) being de most frequent. The overall frequency of HPV-HR was 24.3%, of these 67.4% presented an altered Pap. There was an association between cytological alterations and HPV (p < 0.0001) and years of sexual activity and atypical smear or cervical intraepithelial neoplasia grade I (CIN I) (p = 0.009). 11.9% of young women (21/177) presented atypical smear or CIN I, with 66.7% of cases HPV-HR. Conclusions: These findings alert the vulnerability of these young women who would have a potential risk of viral persistence, CIN and eventually cancer. It is important to emphasize counseling and prevention prior to the regular age of admission to the screening program for cervical cancer. This study was financed by the Universidad de La Frontera through Projects DI15-0047 and DI17-0123.

Genotipificación de VPH en papilomas orales de pacientes con infección por VIH del Hospital San Juan de Dios / HPV genotypes in oral papilloma in Hospital San Juan de Dios HIV (+) patients

Resumen Introducción: Pacientes con infección por VIH presentan mayor riesgo de infecciones orales con genotipos de virus papiloma humano (VPH) de alto riesgo oncogénico (VPH-AR). Objetivo: Determinar los genotipos de VPH en lesiones papilomatosas orales de pacientes con infección por VIH y describir los factores clínicos, histopatológicos y recuento de linfocitos T CD4+ y carga viral asociados. Métodos: Se estudiaron ocho sujetos adultos con infección por VIH y lesiones papilomatosas por VPH. Se extrajo el ADN de la lesión y se detectó el genoma y los genotipos de VPH mediante reacción de polimerasa en cadena e hibridación con el kit comercial HPV 3.5 LCD-Array (Chipron®) y se describieron factores asociados. Resultados: El 63% de los pacientes exhibió más de un genotipo de VPH y 75% de ellos exhibió al menos un genotipo VPH-AR. El genotipo más frecuente fue el VPH 52 (27%), seguido del VPH16 y 56 (18%). El recuento medio de linfocitos T CD4+ en pacientes con al menos un genotipo VPH-AR fue de 330,6 céls/mm3. Conclusiones: Se detectó una mayor frecuencia de infecciones múltiples por VPH, incluido al menos un genotipo de alto riesgo. El genotipo VPH-AR 52 fue el más frecuente. El recuento medio de LT CD4+ en pacientes que presentan al menos un genotipo VPH-AR indica una inmunosupresión moderada. Se requiere aumentar el número de pacientes.

Background: HIV (+) patients have a higher risk of oral infections with high oncogenic risk HPV (HPV-HR). Aim: To determine the HPV genotypes in oral papillomatous lesions in HIV (+) patients and describe the associated factors. Methods: Eight adults HIV (+) subjects with papillomatous HPV lesions were studied. The lesions DNA was extracted and HPV genome and genotypes were detected by PCR and the commercial kit HPV 3.5 LCD-Array Kit (Chipron®) and associated factors were described. Results: 63% of patients exhibited more than one HPV genotype and 75% of them exhibited at least 1 HPV-HR genotype. The most frequent genotype was HPV 52 (27%), followed by HPV 16 and 56 (18%). The mean CD4 T lymphocyte count in patients with at least one HPV-HR genotype was 330.6 cells/mm3. Conclusions: A higher frequency of multiple HPV infections was detected, including at least one high-risk genotype. The genotype HPV-AR 52 was the most frequent. The mean CD4 T lymphocyte count in patients with at least one HPV-HR genotype indicates moderate immunosuppression. It is required to increase the number of patients.

Susceptibilidad genética frente al cáncer bucal por infección del virus del papiloma humano / Genetic susceptibility to oral cancer due to human papillomavirus infection

Resumen Los carcinógenos ambientales y el virus del papiloma humano (VPH) son los principales responsables del cáncer bucal. Los factores de susceptibilidad en el genoma humano desempeñan un papel modulador del riesgo, sin embargo, no todos los individuos expuestos a los carcinógenos padecen cáncer. El objetivo de la presente revisión es describir los principales factores de susceptibilidad genética para cáncer bucal en individuos con infección por VPH. Se realizó una búsqueda sistemática en tres bases de datos en inglés; solo siete artículos cumplieron con los criterios de selección. Se registraron polimorfismos genéticos en tres categorías relacionados con el VPH y que participan en la oncogénesis. Se identificaron tres artículos relativos a la desregulación de los mecanismos de control del ciclo celular, uno relativo a mutaciones en la vía de la apoptosis y tres a polimorfismos en genes de respuesta inflamatoria e inmune. La asociación entre polimorfismos para el desarrollo de cáncer bucal y VPH es evidente, aunque continúa en estudio ya que no siempre el patrón de oncogénesis de las neoplasias bucales está relacionado con el VPH, sino con otros factores ambientales o epigenéticos.

Abstract Environmental carcinogens and human papillomavirus (HPV) are the main responsible factors for oral cancer. Susceptibility factors in the human genome play a risk-modulating role; however not all individuals exposed to these carcinogens suffer from cancer. The purpose of the present review is to describe the main factors of genetic susceptibility to oral cancer due to HPV infection. A systematic search was carried out in three databases in English, with only 7 articles meeting the selection criteria. Genetic polymorphisms are shown in three categories, which are related to HPV and participate in oncogenesis. Three articles related to deregulation of cell cycle control mechanisms were identified, as well as one referring to mutations in the apoptosis pathway and three about polymorphisms in inflammatory and immune response genes. The association of polymorphisms for the development of oral cancer by HPV is evident, although it remains under study. Oral neoplasms’ oncogenesis pattern is not always associated with HPV, but with other environmental or epigenetic factors.

Elevated prevalence of high-risk HPV in healthy oral mucosa of users and nonusers of drugs in Northeastern Brazil

Abstract: Variable rates of HPV infection have been reported in healthy oral mucosa worldwide. The main objective of this study was to detect and genotype HPV infection in users and nonusers of drugs with clinically healthy mucosa from the Northeast Brazil. Samples from 105 patients were amplified using the primers MY09/MY11 and GP5+/GP6+, and genotyping was performed by multiplex-PCR for HPV-6/11, 16 and 18. A total of 81.9% samples were positive. Among drug users, 84.5% presented the virus and 20.4% showed multiple infections. Among non-drug users, 78.7% were positive and 13.5% had multiple infections. Limited information is available on oral HPV in Brazilian population, especially for drug users, and our results showed higher HPV infection rates in both users and nonusers of drugs. More studies and researches focused on drug users including factors like sexual behavior, nutrition and cultural habits are necessary to enhance the comprehension of this relationship, and develop preventive strategies.

Lower genital tract infections in young female juvenile idiopathic arthritis patients

Abstract Background: To evaluate human papillomavirus (HPV), Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) infections in juvenile idiopathic arthritis (JIA) patients. Methods: After exclusion, 33 female adolescent and young JIA patients (ILAR criteria) and 28 healthy controls were selected for this study. Demographic data, gynecological, sexual function, cervical cytology and histological abnormalities were evaluated. JIA clinical/laboratorial parameters and treatment were also assessed. HPV-DNA, CT-DNA and NG-DNA testing in cervical specimens were performed by Hybrid Capture 2 assays. Results: The mean current age was similar in JIA patients and controls (23.3 ± 6.24 vs. 26.1 ± 6.03 years, p = 0.09). The frequencies of sexual intercourse (76% vs. 89%, p = 0.201) and abnormal cervical cytology (24% vs. 11%, p = 0.201) were similar in JIA compared to controls. The higher frequency of HPV infection in JIA patients than controls (30% vs. 11%, p = 0.155) did not reach statistical significance. CT (0% vs. 7%, p = 0.207) and NG infections (0% vs. 4%, p = 0.459) were also alike in both groups. Further evaluation of JIA patients with abnormal and normal cervical cytology showed that the former group had a higher frequency of HPV infection (87% vs. 12%, p = 0.0002) with a low frequency of HPV vaccination (0% vs. 8%, p = 1.0). No differences were evidenced between these two JIA groups regarding demographic data, sexual function and clinical/laboratorial parameters. The frequencies of methotrexate (p =0.206) and biological agent use (p =0.238) were similar in both JIA groups. Conclusions: To our knowledge, this was the first study to assess lower genital infections in JIA patients allowing the identification of HPV as main cause of cervical dysplasia. Methotrexate and biological agents do not seem to increase risk of lower genital tract infections in JIA patients.

Incidence, psychosocial burden, and economic impact of genital warts in Mexico / Incidencia, carga psicosocial e impacto económico de las verrugas genitales en México

Abstract: Objective: To estimate the burden of genital warts (GW) in Mexico. Materials and methods: We estimated the annual incidence of GW based on data reported by specialist physicians. We also assessed GW treatment practices, the average cost of treatment, and the psychosocial burden of GW among patients. Results: The annual incidence of GW in Mexico was estimated to be 547 200 cases. Treatment procedures vary by specialist and patient gender. The estimated annual cost was $195 million USD. The psychosocial impact of GW was slightly greater in males than females. Conclusions: This is the first evaluation of the burden of GW in Mexico. Our data suggest that GW are common, with significant health-related costs and psychosocial impact.

Resumen: Objetivo: Estimar la carga por verrugas genitales (VG) en México. Material y métodos: Estimamos la incidencia anual de VG, con base en información proporcionada por médicos especialistas y el manejo de las VG, así como el costo promedio del tratamiento y la carga psicosocial de las VG. Resultados: La incidencia anual de VG en México fue de 547 200 casos. Los tratamientos variaron según la especialidad y el sexo del paciente. El costo anual por VG fue de $195 millones de dólares estadounidenses. El impacto psicosocial de las VG es ligeramente mayor en hombres que en mujeres. Conclusiones: Esta es la primera evaluación de la carga de VG en México. Los datos sugieren que las VG son frecuentes, tienen costos relacionados con salud e impactos psicosociales significativos.

Prevalence of Trichomonas vaginalis and Human papillomavirus in female sex workers in Central Veracruz, Mexico / Prevalencia de Trichomonas vaginalis y virus del papiloma humano en mujeres trabajadoras sexuales en el centro de Veracruz, México

Female sex workers (FSWs) have been considered a key population for sexually transTrichomonas mitted infections (STIs); therefore, they are periodically screened as a requirement to obtain a work card. However, there is insufficient epidemiological data on STIs among FSWs in Mexico. The detection of Trichomonas vaginalis is limited to microscopic studies and the molecular screening of Human papillomavirus (HPV) is only done to women 35 years of age and older. The objective of this study was to determine the prevalence of T. vaginalis and HPV infections in FSWs in the city of Orizaba, Veracruz, Mexico. Samples from 105 FSWs were obtained by cervical swab and analyzed. The identification of T. vaginalis and HPV was performed by molecular methods. HPV DNA was identified in 5.71% of the samples with the presence of HPV16, HPV18, and HPV58. A percentage of 25.7% samples were positive for T. vaginalis for optical microscopy and 23.8% for PCR. The results of the study indicate the need to incorporate more sensitive methods for the timely diagnosis of STIs as well as comprehensive health promotion programs directed to the most vulnerable groups among FSWs.

Las mujeres trabajadoras sexuales (MTS) han sido consideradas una población clave para las infecciones de transmisión sexual (ITS), por ello son examinadas periódicamente como requisito para obtener una tarjeta de trabajo. Sin embargo, no existen datos epidemiológicos suficientes sobre las ITS en las MTS en México. La detección de Trichomonas vaginalis se limita a los estudios microscópicos, y el cribado molecular del virus del papiloma humano (Human papillomavirus: HPV) solo se realiza en las mujeres de 35 años o mayores. El objetivo de este estudio fue determinar la prevalencia de T. vaginalis e infecciones por HPV en las MTS de la ciudad de Orizaba, Veracruz, México. Se analizaron 105 muestras de las MTS, obtenidas mediante frotis cervical. La identificación de T. vaginalis y HPV se realizó por métodos moleculares. El ADN del HPV se identificó en el 5,71% de las muestras, con la presencia de HPV16, HPV18 y HPV58. El 25,7% de las MTS fueron positivas para T. vaginalis por microscopia óptica el 23,8% por PCR. Los resultados del estudio indican la necesidad de incorporar métodos más sensibles para el diagnóstico oportuno de ITS y programas integrales de promoción de la salud en los grupos más vulnerables, entre las MTS.

Detección del virus del papiloma humano en muestras obtenidas mediante técnica de autotoma en un grupo de universitarias peruanas / Detection of the human papillomavirus in samples obtained by self-collection technique in a group of peruvian college students

RESUMEN Estudio transversal que determinó la frecuencia y genotipos del virus del papiloma humano de alto riesgo (VPH-AR) a través de la técnica de autotoma en un grupo de universitarias de Lima. Participaron 221 estudiantes y se detectó el ADN del VPH-AR con el método de reacción en cadena de la polimerasa (PCR). La frecuencia del VPH-AR en las participantes fue de 43,4%; de este grupo se encontraron los genotipos VPH 16 en el 15,6% y VPH 18 en el 4,2% y otros VPH-AR en el 80,2%. Se concluye que la frecuencia del VPH-AR es mayor en el grupo de universitarias de este estudio en comparación a investigaciones nacionales previas.

ABSTRACT Cross-sectional study that determined the frequency and the genotypes of the (HR-HPV) high-risk human papillomavirus through the self-collection technique in a group of college students of Lima. Two hundred twenty-one (221) students participated and the DNA of the HR-HPV was detected with polymerase chain reaction (PCR). The frequency of HR-HPV in participants was 43.4%; of this group, genotype HPV 16 was present in 15.6%, HPV 18 in 4.2%, and another HR-HPV in 80.2%. We can conclude that the frequency of HR-HPV is greater in the group of college students of this study in comparison with previous national investigations.

Epidemiology and biology of cutaneous human papillomavirus

Cutaneous human papillomaviruses (HPVs) include β- and γ-HPVs, in addition to a small fraction of α-HPVs. β-HPVs were first isolated from patients with the rare genetic disorder Epidermodysplasia verruciformis, and they are associated with the development of nonmelanoma skin cancer at sun-exposed skin sites in these individuals. Organ transplant recipients also have greater susceptibility to β-HPV infection of the skin and an increased risk of developing nonmelanoma skin cancer. In both immunosuppressed and immunocompromised individuals, cutaneous HPVs are ubiquitously disseminated throughout healthy skin and may be an intrinsic part of the commensal flora. Functional analysis of E6 and E7 proteins of specific cutaneous HPVs has provided a mechanistic comprehension of how these viruses may induce carcinogenesis. Nevertheless, additional research is crucial to better understand the pathological implications of the broad distribution of these HPVs.

A pilot study on the identification of human papillomavirus genotypes in tongue cancer samples from a single institution in Ecuador

The relationship between human papillomavirus (HPV) and oropharyngeal squamous cell carcinoma has been established. However, data from Ecuador is limited. The objective of this study was to characterize HPV infection in Ecuadorian patients with tongue cancer. Fifty-three patients with tongue cancer treated at the tertiary referral center Sociedad de Lucha Contra el Cancer (SOLCA), Guayaquil, between 2006 and 2011 were identified. Linear Array® HPV genotyping was used to identify the presence and types of HPV on formalin-fixed paraffin-embedded biopsy samples from these patients with tongue cancer. HPV was identified in 42% (n=22) and high-risk (HR) HPV in 17% (n=9), with 18 different HPV types identified. The most common types were the HR HPV 33 (14%) and low-risk HPV 67 (14%), followed by the HR HPV 58. More than one HPV type was identified in 27.3% of cases. HPV 33 was frequently associated with other HPV types. No statistically significant differences in gender (P=0.58) and age (P=0.12) were observed between HPV-positive and HPV-negative cases. HPV was identified in almost half of the tongue cancer samples, with subtypes 33 and 67 being the most common. This suggested that HPV played an important role in this disease in the population studied. Given these results, current HPV vaccines may not be as effective in reducing tongue cancer rates in this population.

Human papillomavirus infection and cervical cancer precursor lesions in women living by Amazon rivers: investigation of relations with markers of oxidative stress / Infecção por papilomavírus humano e lesões precursoras do câncer cervicouterino em Ribeirinhas da Amazônia: avaliação da relação com marcadores de estresse oxidativo

ABSTRACT Objective To investigate the relation between oxidative stress markers, human papillomavirus infection and cervical cancer precursor lesions. Methods The study comprised women aged 14 to 60 years living in communities located by Amazon rivers in the state of Pará (Itaituba, Limoeiro do Ajuru and Bragança, 126, 68 and 43 women respectively). Papanicolau smears and polymerase chain reaction tests for human papillomavirus DNA detection were performed. Blood samples were collected to test malondialdehyde, total and oxidized glutathione levels. Results Malondialdehyde, total and oxidized glutathione concentrations did not differ significantly (p>0.05) between women with and without low-grade squamous intraepithelial lesions across communities. Malondialdehyde levels (8.02nmols/mL) were almost five times higher in human papillomavirus-positive compared to human papillomavirus-negative women (1.70nmols/mL) living in Itaituba (statistically significant difference; p<0.05). Malondialdehyde levels did not differ significantly (p>0.05) between human papillomavirus-positive and human papillomavirus-negative women living in remaining communities. Significant (p<0.05) differences in total glutathione levels between human papillomavirus-positive and human papillomavirus-negative women (8.20μg/mL and 1.47μg/mL, respectively) were limited to those living in Bragança. Conclusion Malondialdehyde and total glutathione levels were significantly associated with human papillomavirus infection. However, lack of similar associations with squamous lesions suggest oxidative stress alone does not explain correlations with cervical carcinogenesis. Other factors may therefore be involved.

RESUMO Objetivo Avaliar a relação de marcadores de estresse oxidativo com a infecção pelo papilomavírus humano e as lesões precursoras do câncer cervical. Métodos Foram incluídas mulheres ribeirinhas da Amazônia (a saber: 126 do município de Itaituba, 68 de Limoeiro do Ajuru e 43 de Bragança), todas do Estado do Pará, de 14 a 60 anos. Foram realizados testes Papanicolau e de reação em cadeia de polimerase para a detecção de DNA do papilomavírus humano. Coleta de amostras de sangue foi realizada para a avaliação do malondialdeído e a determinação da glutationa total e oxidada. Resultados As concentrações oxidativas do malondialdeído e os teores de glutationa total e oxidada não apresentaram diferença significativa (p>0,05) nas mulheres com e sem lesão intraepitelial escamosa de baixo grau, em todas as comunidades pesquisadas. Mulheres com papilomavírus humano em Itaituba apresentaram níveis de malondialdeído (8,02nmols/mL) quase cinco vezes maior quando relacionadas àquelas sem o HPV (1,70nmols/mL), com diferença estatística significativa (p<0,05). As outras comunidades não mostraram diferença estatística significativa entre as concentrações de malondialdeído nas mulheres com e sem papilomavírus humano (p>0,05). Os teores de glutationa total mostraram diferença significativa (p<0,05) somente em Bragança naquelas com (8,20µg/mL) e sem a infecção pelo papilomavírus humano (1,47µg/mL). Conclusão Houve associação significativa da resposta oxidativa do malondialdeído e da glutationa total com a infecção pelo papilomavírus humano, porém não houve diferença quando associada à lesão escamosa, sugerindo que o estresse oxidativo isoladamente não explica a relação com a carcinogênese do colo uterino, que deve ser influenciada ainda por outros fatores.

Epidemiología de la infección oral por VPH en sujetos jóvenes sanos / Epidemiology of oral HPV infection in young healthy individuals

Resumen Introducción: La infección por VPH es la infección viral de transmisión sexual más frecuente, y se encuentra asociada a diversas neoplasias. Objetivo: Describir la epidemiología, historia natural y factores de riesgo asociados a la infección oral por VPH en adultos jóvenes asintomáticos. Métodos: Se realizó un estudio prospectivo de corte longitudinal, que incluyó sujetos sin patología oral, a los que se les tomó una muestra de la mucosa bucal. A todos los sujetos con resultados positivos se les realizó un nuevo muestreo seis meses después. Se identificó la presencia del virus por RPC; los datos demográficos y de conducta sexual fueron obtenidos con una encuesta que respondieron sin intervención del investigador. Resultados: Se recolectaron 102 muestras de sujetos de 18-26 años de edad, 60 (58,8%) correspondieron al sexo masculino. La prevalencia del virus fue de 6,9%; todos los sujetos positivos tenían vida sexual. Las relaciones sexuales entre personas del mismo sexo fue la única variable asociada a la presencia del virus (p < 0,05). A los seis meses, todos los sujetos habían eliminado al virus. Conclusión: La infección oral por VPH es transitoria y está asociada a relaciones sexuales entre personas del mismo sexo, principalmente mujeres que tienen sexo con mujeres.

Background: HPV infection is the most common sexually transmitted viral infection, and is associated with several neoplasms. Aim: To describe the epidemiology, natural history and risk factors associated with oral HPV infection in asymptomatic young adults. Methods: A prospective and longitudinal study was conducted, including subjects without oral pathology, who were sampled from the oral mucosa. All subjects with positive results were re-sampled 6 months later. The presence of HPV was identified by PCR. Demographic and sexual behavior data were obtained with a survey that was responded without the intervention of the researcher. Results: 102 samples were collected from subject of 18-26 years old, 60 (58.8%) were male. The prevalence of the virus was 6.9%; all positive subjects had active sexual life. Same-gender relationships were the only variable associated with the presence of the virus (p < 0.05). At six months all subjects had eliminated the virus. Conclusion: Oral HPV infection is transient and is associated to same-gender relationships, mainly women who have sex with women.

Detection of High-Risk Human Papillomavirus in Cervix Sample in an 11. 3-year Follow-Up after Vaccination against HPV 16/18 / Detecção de papilomavírus humano de alto risco em amostras de colo uterino em 11. 3 anos de acompanhamento após vacinação contra HPV 16/18

Abstract Purpose the aim of this study was to evaluate the pattern of human papillomavirus (HPV) detection in an 11.3-year post-vaccination period in a cohort of adolescent and young women vaccinated or not against HPV 16/18. Methods a subset of 91 women from a single center participating in a randomized clinical trial (2001-2010, NCT00689741/00120848/00518336) with HPV 16/18 AS04- adjuvanted vaccine was evaluated. All women received three doses of the HPV vaccine (n = 48) or a placebo (n = 43), and cervical samples were collected at 6-month intervals. Only in this center, one additional evaluation was performed in 2012. Up to 1,492 cervical samples were tested for HPV-DNA and genotyped with polymerase chain reaction (PCR). The vaccine group characteristics were compared by Chi-square or Fisher exact or Mann-Whitney test. The high-risk (HR)-HPV 6-month-persistent infection rate was calculated. The cumulative infection by HPV group was evaluated by the Kaplan-Meier method and the log-rank test. Results the cumulative infection with any type of HPV in an 11.3-year period was 67% in the HPV vaccine group and 72% in the placebo group (p = 0.408). The longitudinal analysis showed an increase of 4% per year at risk for detection of HR-HPV (non-HPV 16/ 18) over time (p = 0.015), unrelated to vaccination. The cumulative infection with HPV 16/18 was 4% for the HPV vaccine group and 29% for the placebo group (p = 0.003). There were 43 episodes of HR-HPV 6-month persistent infection, unrelated to vaccination. Conclusions this study showed themaintenance of viral detection rate accumulating HR-HPV (non-HPV-16-18) positive tests during a long period post-vaccination, regardless of prior vaccination. This signalizes that the high number of HPV-positive testsmay be maintained after vaccination.

Resumo Objetivos avaliar o padrão de detecção do papilomavírus humano (HPV) em um período de 11.3 anos após a vacinação em uma coorte de adolescentes e mulheres jovens vacinadas ou não contra HPV 16/18. Métodos avaliou-se um subgrupo de 91 mulheres de um único centro, participantes de ensaio clínico randomizado (2001-2010, NCT00689741/00120848/00518336) com a vacina contra HPV 16/18 com adjuvante AS04. Todas as mulheres receberam três doses de vacina contra HPV (n = 48) ou placebo (n = 43), e tiveram amostras cervicais coletadas em intervalos de 6 meses. Somente neste centro, uma avaliação adicional foi realizada em 2012. Um total de 1.492 amostras cervicais foram testadas para DNA-HPV e genotipadas com reação em cadeia da polimerase (RCP). As características dos grupos de vacina contra HPV ou placebo foram comparadas pelo teste de Qui-quadrado ou teste exato de Fisher ou teste de Mann-Whitney. A infecção persistente por 6meses pelo HPV de alto risco (AR) foi calculada. A infecção cumulativa por grupo foi avaliada pelo método de Kaplan-Meier e pelo teste log-rank. Resultados a infecção cumulativa com qualquer tipo de HPV em11.3 anos foi de 67% no grupo vacina contra HPV e de 72% no grupo placebo (p = 0,408). A análise longitudinal mostrou um aumento de 4% ao ano no risco de detecção de HR-HPV (não-HPV 16/18) ao longo do tempo (p = 0,015), não relacionado com a vacinação. A infecção cumulativa com HPV 16/18 foi de 4% para o grupo vacina contra HPV e 29% para o grupo placebo (p = 0,003). Houve 43 episódios de infecção persistente por 6 meses por HR-HPV, não relacionados com a vacinação. Conclusões este estudo mostrou a manutenção da taxa de detecção viral, acumulando testes positivos de HR-HPV (não HPV-16-18) durante longo período pósvacinação, independentemente da vacinação prévia. Isto sinaliza que a alta positividade dos testes de HPV pode ser mantida após a vacinação.