Human Papillomavirus (HPV) Infection and Risk Behavior in Vaccinated and Non-Vaccinated Paraguayan Young Women.

Cervical cancer is a global health concern and ranks fourth among the most prevalent cancers in women worldwide. Human papillomavirus (HPV) infection is a known precursor of cervical cancer and preventive measures include prophylactic vaccines. This study focused on sexually active Paraguayan women aged 18-25 years, exploring the intersection of HPV vaccination and sexual behavior. Among 254 participants, 40.9% received the Gardasil-4 vaccine, with no significant differences in sexual behavior between the vaccinated and unvaccinated sexually active groups. However, a notable decrease in the prevalence of HPV among the vaccinated women highlights the efficacy of this vaccine in reducing infections. The prevalence of any HPV type was 37.5% in vaccinated participants compared to 56.7% in unvaccinated participants (p = 0.0026). High-risk HPV types showed a significant difference, with a prevalence of 26.0% in vaccinated women compared with 52.7% in unvaccinated women (p < 0.001). Although a potential decline in genital warts was observed among the vaccinated individuals, statistical significance (p = 0.0564) was not reached. Despite the challenges in achieving high vaccination coverage, the observed reduction in HPV prevalence underscores the importance of ongoing monitoring, healthcare professional recommendations, and comprehensive risk management. These findings contribute to dispelling concerns about HPV vaccination influencing sexual behavior, advocating further large-scale research to explore the impact of vaccines on various HPV types and potential cross-protection.

High prevalence and co-infection of high-risk Human Papillomavirus genotypes among unvaccinated young women from Paraguay.

Paraguay launched a human papillomavirus (HPV) vaccination program in 2013, so virological surveillance is important for measuring the impact of HPV vaccines. This study aimed to estimate the type-specific HPV frequency in unvaccinated sexually active women aged 18-25 years in the metropolitan area of Asuncion as a baseline for monitoring the HPV vaccination program. This study included 208 women, attending the Central Laboratory of Public Health between May 2020 and December 2021, were invited for testing through social networks and flyers at local health centers and higher education institutes. All participants who agreed to contribute to the study signed a free, prior, and informed consent form and answered a questionnaire that included basic demographic data and determining factors of HPV infection. Human papillomavirus detection and genotyping were conducted using the CLART HPV2 test (Genomica, Madrid, Spain) that allows the individual identification of 35 genotypes. 54.8% women were positive for any HPV type, with 42.3% positive for high-risk HPV (HR-HPV) types. Several factors were associated with HPV detection including the number of sexual partners, new sexual partners, non-use of condoms, and history of other sexual infections. Moreover, multiple infections were identified in 43.0% of the young women. We detected 29 different viral types present in both single and multiple infections. HPV-58 was the most commonly detected HPV type (14.9%), followed by HPV-16, HPV-51, and HPV-66 (12.3%). We estimated the prevalence of bivalent (16/18), quadrivalent (6/11/16/18), and nonavalent (6/11/16/18/31/33/45/52/58) vaccine types to be 8.2%, 13%, and 38%, respectively. These results reinforce the importance of surveillance studies and provide the first data regarding circulating HPV genotypes in the unvaccinated population in Paraguay, thus generating a baseline to compare future changes in the overall and type-specific HPV prevalence after HPV vaccination.

Implementing HPV testing in 9 Latin American countries: The laboratory perspective as observed in the ESTAMPA study.

Background: Replacement of cytology screening with HPV testing is recommended and essential for cervical cancer elimination. HPV testing for primary screening was implemented in 12 laboratories within 9 Latin American countries, as part of the ESTAMPA cervical cancer screening study. Our observations provide information on critical operational aspects for HPV testing implementation in diverse resource settings. Methods: We describe the implementation process of HPV testing in ESTAMPA, focusing on laboratory aspects. We assess the readiness of 12 laboratories to start HPV testing and their continuity capacity to maintain good quality HPV testing until end of recruitment or up to December 2021. Readiness was based on a checklist. Information from the study database; regular meetings and monitoring visits; and a questionnaire on laboratory operational aspects sent in May 2020 were used to assess continuity capacity. Compliance with seven basic requirements (readiness) and eight continuity requirements (continuity capacity) was scored (1 = compliant, 0 = not compliant) and totaled to classify readiness and continuity capacity as very limited, limited, moderate or high. Experiences, challenges, and enablers of the implementation process are also described. Results: Seven of 12 laboratories had high readiness, three moderate readiness, and of two laboratories new to HPV testing, one had limited readiness and the other very limited readiness. Two of seven laboratories with high readiness also showed high continuity capacity, one moderate continuity capacity, and the other four showed limited continuity capacity since they could not maintain good quality HPV testing over time. Among three laboratories with moderate readiness, one kept moderate continuity capacity and two reached high continuity capacity. The two laboratories new to HPV testing achieved high continuity capacity. Based on gained expertise, five laboratories have become part of national screening programs. Conclusion: High readiness of laboratories is an essential part of effective implementation of HPV testing. However, high readiness is insufficient to guarantee HPV testing high continuity capacity, for which a "culture of quality" should be established with regular training, robust monitoring and quality assurance systems tailored to local context. All efforts to strengthen HPV laboratories are valuable and crucial to guarantee effective implementation of HPV-based cervical screening.

Distribución de virus de papiloma humano de alto riesgo oncogénico y otras infecciones de transmisión sexual en mujeres paraguayas con y sin virus de la inmunodeficiencia humana

El virus de papiloma humano de alto riesgo oncogénico (VPH-AR) es causa necesaria pero no suficiente para la ocurrencia de cáncer de cuello uterino (CCU). Mujeres portadoras del virus de inmunodeficiencia humana (VIH) presentan mayor riesgo de desarrollar lesiones precursoras del cáncer de cuello de útero, por ello, el objetivo del presente trabajo prospectivo de corte transversal fue determinar la frecuencia de VPH-AR y otras infecciones de transmisión sexual-ITS (condilomas, sífilis, virus del herpes simple, gonorrea, citomegalovirus, hepatitis B) en 218 mujeres con y sin VIH que acudieron al Programa Nacional de Lucha contra el SIDA (PRONASIDA) desde julio 2017 hasta marzo 2021. Se encontró que 16/54 (29,6%) mujeres VIH-positivas presentaron infección por VPH-AR en comparación a 41/164 (25%) mujeres VIH-negativas (p>0,05). En relación a la edad, mujeres VIH positivas presentaron una frecuencia comparable de infección por VPH-AR (30 años 30,2%), a diferencia de mujeres VIH negativas donde hubo una disminución significativa de la infección por VPH-AR luego de los 30 años (30 años 18,8%, p= 0,028). Esto podría explicarse por la inmunosupresión observada en mujeres VIH positivas que podría favorecer infecciones persistentes, sugiriendo que deben ser controladas más cercanamente. Además, se observó mayor frecuencia de otras ITS en mujeres VIH positivas (29,6% vs 15,8%, p=0,026), lo cual sugiere que aparte del monitoreo más cercano, es fundamental fortalecer la educación sobre factores de riesgo para la ITS sobre todo VPH y VIH, así como la realización de prevención primaria por vacunación contra el VPH.

High-risk human papillomavirus (HPV-HR) is a necessary but not sufficient cause for cervical cancer (CC). Women carriers of human immunodeficiency virus (HIV) present an increased risk for the development of cervical cancer precursor lesions, therefore, the objective of the present prospective cross-sectional study was to determine the frequency of HPV-HR and other sexually transmitted infections-STIs (condylomas, syphilis, herpes simplex virus, gonorrhoea, cytomegalovirus, hepatitis B) in 218 women with and without HIV who attended the Ministry of Health from July 2017 to March 2021. It was found that 16/54 (29.6%) HIV-positive women had HPV infection compared to 41/164 (25%) HIV-negative women (p>0.05). In relation to age, HIV-positive women had a comparable frequency of HPV infection (30 years 30.2%), unlike HIV-negative women whom above 30 years of age presented a significant decrease in HPV-AR infection (30 years 18.8%, p:0.028). This could be explained by the immunosuppression observed in HIV-positive women which could favour persistent infections, suggesting that they should be controlled more closely. In addition, other STIs were observed to be more frequent in HIV-positive women (29.6% vs 15.8%, p:0.026), which suggests that apart from closer monitoring, it is essential to strengthen education on risk factors for STIs, especially HPV and HIV, as well as the implementation of primary prevention by vaccination against HPV.

Multicentric study of cervical cancer screening with human papillomavirus testing and assessment of triage methods in Latin America: the ESTAMPA screening study protocol.

INTRODUCTION: Human papillomavirus (HPV) testing is replacing cytology in primary screening. Its limited specificity demands using a second (triage) test to better identify women at high-risk of cervical disease. Cytology represents the immediate triage but its low sensitivity might hamper HPV testing sensitivity, particularly in low-income and middle-income countries (LMICs), where cytology performance has been suboptimal. The ESTAMPA (EStudio multicéntrico de TAMizaje y triaje de cáncer de cuello uterino con pruebas del virus del PApiloma humano; Spanish acronym) study will: (1) evaluate the performance of different triage techniques to detect cervical precancer and (2) inform on how to implement HPV-based screening programmes in LMIC. METHODS AND ANALYSIS: Women aged 30-64 years are screened with HPV testing and Pap across 12 study centres in Latin America. Screened positives have colposcopy with biopsy and treatment of lesions. Women with no evident disease are recalled 18 months later for another HPV test; those HPV-positive undergo colposcopy with biopsy and treatment as needed. Biological specimens are collected in different visits for triage testing, which is not used for clinical management. The study outcome is histological high-grade squamous intraepithelial or worse lesions (HSIL+) under the lower anogenital squamous terminology. About 50 000 women will be screened and 500 HSIL+ cases detected (at initial and 18 months screening). Performance measures (sensitivity, specificity and predictive values) of triage techniques to detect HSIL+ will be estimated and compared with adjustment by age and study centre. ETHICS AND DISSEMINATION: The study protocol has been approved by the Ethics Committee of the International Agency for Research on Cancer (IARC), of the Pan American Health Organisation (PAHO) and by those in each participating centre. A Data and Safety Monitoring Board (DSMB) has been established to monitor progress of the study, assure participant safety, advice on scientific conduct and analysis and suggest protocol improvements. Study findings will be published in peer-reviewed journals and presented at scientific meetings. TRIAL REGISTRATION NUMBER: NCT01881659.

Caracterización epidemiológica y molecular de la Hepatitis C en la población que acudió al Laboratorio Central de Salud Pública, Paraguay 2013-2018 / Epidemiological and molecular characterization of Hepatitis C in the population that went to the Central Laboratory of Public Health, Paraguay 2013-2018

Introducción: Las hepatitis causadas por el virus de la hepatitis C (VHC) se han transformado en uno de los principales problemas de enfermedades infecciosas emergentes, responsables del 80% de las hepatitis crónicas con posible evolución a cirrosis o carcinoma hepatocelular y ocasionando un alto costo para el sistema de salud. Objetivo: Describir el perfil epidemiológico y los genotipos del VHC en pacientes que acudieron al Laboratorio Central de Salud Pública (LCSP). Materiales y métodos: Estudio descriptivo; se incluyeron 162 pacientes con infección por Hepatitis C referidos al LCSP entre el 2013 y 2018, para seguimiento y/o genotipificación. Se les realizó la amplificación del genoma mediante la técnica reacción en cadena de la polimerasa en tiempo real previa transcripción reversa (RT-PCR). A una submuestra con PCR detectable y carga viral >500 UI/ml se determinó el genotipo(n=52). Resultados: La media de edad fue de 44,2 ±15,6 años, el 52,5% eran hombres. El 8,02% presentaron carga viral alta, 32,09 % baja y 59,87 % indetectable. La distribución de genotipos fue la siguiente: 61,5 % genotipo 1 (28,1% 1a, 53,1% 1b y 18,8% genotipo 1 sin subtipificación), 15,4% genotipo 2, 15,4% genotipo 3 y 7,7% genotipo 4. Conclusiones: El presente trabajo muestra la importancia de la implementación de técnicas moleculares aplicadas a la vigilancia epidemiológica de nuestro país de manera a establecer programas de detección temprana y seguimiento adecuado de los pacientes, ya que la caracterización genotípica ayuda a determinar lasestrategias terapéuticas más adecuadas y predecir la respuesta antiviral. Se confirma que el genotipo 1 es el que circula con mayor frecuencia, con alto predominio del subtipo 1b. Palabras Clave: Biología Molecular, Hepatitis C, Genotipo, Epidemiología Molecular, Paraguay.

Introduction: Hepatitis caused by the hepatitis C virus (HCV) has become one of the main problems of emerging infectious diseases, responsible for 80% of chronic hepatitis with possible evolution to cirrhosis or hepatocellular carcinoma and causing a high cost for the health system. Objective: To describe the epidemiological profile and the genotypes of HCV in patients who attended the Central Public Health Laboratory (LCSP). Materials and methods: Descriptive study; included 162 patients with Hepatitis C infection referred to the LCSP between 2013 and 2018, for follow-up and / or genotyping. Genome amplification was performed using the polymerase chain reaction technique in real time prior to reverse transcription (RT-PCR). To a subsample with detectable PCR and viral load> 500 IU / ml, the genotype was determined (n = 52). Results: The mean age was 44.2 ± 15.6 years, 52.5% were men. The 8.02% had high viral load, 32.09% low and 59.87% undetectable. The genotype distribution was as follows: 61.5% genotype 1 (28.1% 1a, 53.1% 1b and 18.8% genotype 1 without subtyping), 15.4% genotype 2, 15.4% genotype 3 and 7.7% genotype 4. Conclusions: The present work shows the importance of the implementation of molecular techniques applied to the epidemiological surveillance of our country in order to establish programs of early detection and adequate monitoring of patients, since genotypic characterization helps to determine the most appropriate therapeutic strategies and predict the antiviral response. It is confirmed that genotype 1 is the one that circulates more frequently, with a high predominance of subtype 1b. Keywords: Molecular Biology, Hepatitis C, Genotype, Molecular Epidemiology, Paraguay.

Detección y tipificación del Virus Papiloma Humano en el marco del tamizaje virológico para la detección de lesiones del cuello uterino en Asunción, Paraguay / Detection and typing of Human Papilloma Virus during a virological screening for detection of cervical lesions in Asunción, Paraguay

En Paraguay la incidencia de cáncer de cuello uterino (CCU) es superior a las observadas en otros países de la región. El agente etiológico asociado al CCU es el virus papiloma humano (VPH), esencialmente tipos de alto riesgo oncogénicos. El objetivo es describir aspectos epidemiológicos de la infección genital por el virus papiloma humano de alto riesgo (VPH-AR) en mujeres de 25 a 64 años que consultaron en servicios de Patología Cervical del MSPyBS, de mayo a diciembre de 2013. Se utilizó el Cobas 4800 HPV Test (Roche) que permite la detección individual de VPH-16 y VPH-18 y un pool de otros VPH-AR que incluye 12 genotipos de alto riesgo. Los otros VPH-AR fueron tipificados por hibridación reversa en línea (RLB). Entre las 495 mujeres incluidas, se detectaron 72 casos positivos (14,5%) de VPH-AR. Se identificaron 19 tipos virales; siendo el más frecuente VPH-16 (2,1%), seguido del VPH-31, 33, 58 y 66; el VPH-18 aparece en sexto lugar. Este trabajo aporta los primeros datos sobre la implementación de técnicas moleculares para detección y tipificación de VPH como parte del sistema de salud pública de Paraguay. El predominio de VPH-16, confirma su amplia circulación a nivel mundial y dado su mayor potencial oncogénico, representa una alerta a considerar, en especial en las mujeres mayores de 30 años portadoras de una infección persistente. Estos resultados apoyan la importancia de la implementación criteriosa y la utilización apropiada de las pruebas moleculares actualmente disponibles para la prevención y control del CCU(AU)

Desempeño de una prueba rápida para el diagnóstico de sífilis en mujeres puérperas / Performance of a rapid test for the diagnosis of syphilis inpuerperal women

La sífilis, por su impacto en la salud materno infantil, demanda prevención y tratamiento de calidad. Las pruebas rápidas son herramientas adecuadas de detección con una sensibilidad entre 84% a 97% y especificidad de 84% o más. En Paraguay, las pruebas rápidas han sido validadas e incorporadas a los programas de salud pero no existen estudios que hayan evaluado su desempeño, por lo que este estudio observacional descriptivo tiene por objetivo evaluar el desempeño de una prueba rápida comercial para el diagnóstico de sífilis en comparación a la Hemaglutinación Indirecta del Treponema (TPHA) en muestras de mujeres puérperas remitidas al Laboratorio Central de Salud Pública por hospitales de parto del Municipio Capital, y los departamentos: Central, Concepción y Alto Paraná entre 2011 y 2012. De 334 muestras evaluadas, 108 fueron positivas por las dos pruebas (test rápido y TPHA), y 182 negativas por ambas. En el resto, los resultados fueron discordantes (23 positivos con la prueba rápida y negativos por TPHA y 21 negativos por test rápido y positivos por TPHA). La sensibilidad de la prueba rápida comparada con TPHA fue 83,72% (IC95%: 75,96-89,42%) y la especificidad de 88,78% (IC95%: 83,45-92,61%).

Due to its impact on maternal and child health, syphilis demands quality in preventionand treatment. The rapid test is an appropriate screening tool with a sensitivity of 84-97%and specificity from 84%. In Paraguay, rapid tests have been validated and incorporatedinto health programs. The performance of a rapid test for syphilis was evaluated comparedwith Treponema particle agglutination assay (TPHA) in samples of puerperal women sent tothe Central Laboratory of Public Health from birthing hospitals of the Capital City, and threedepartments: Central, Concepción and Alto Paraná between 2011 and 2012. Of the 334samples tested, 108 were positive by both tests (rapid test and TPHA test), and 182 werenegative by both. The rest presented discordant results (23 positive with the rapid test andTPHA negative and 21 negative by rapid test and positive for TPHA). The sensitivity of therapid test compared with the TPHA was 83.72% (95% CI 75.96 to 89.42%) and specificity88.78% (95% CI 83.45 to 92.61).

Detección molecular del papilomavirus humano de alto riesgo en el seguimiento de mujeres tratadas por lesión escamosa intraepitelial / Molecular detection of high risk human papillomavirus in following women treated for squamous intraepithelial lesion

El cáncer de cuello uterino (CCU) es la primera causa de muerte por cáncer en mujeres en países en vías de desarrollo. La infección persistente por el virus papiloma humano (VPH) es un factor necesario en lesiones preneoplásicas y CCU. La citología cervicovaginal es el método mayormente utilizado para detectar el CCU y su uso combinado con la de detección de ADN viral seis meses post-tratamiento aumenta la efectividad para identificar mujeres tratadas con riesgo de lesión residual/recidiva. El objetivo fue describir la frecuencia de VPH de alto riesgo (AR) en mujeres tratadas por lesión escamosa intraepitelial (SIL) que acudieron al Servicio de Patología Cervical del Hospital San Pablo de enero-diciembre/2014. Se realizó un estudio descriptivo de corte transverso, que incluyó 80 pacientes que acudieron al servicio para control post-tratamiento por SIL. Se utilizó Cobas 4800 HPV Test (Roche) para la detección individual de VPH-16 y 18, y un pool de 10 VPH-AR (31,33,35,39,45,51,52,56,58,59) y dos de “probable” alto riesgo (66,68). Se encontró infección viral en el 7,5% (6/80) de las pacientes tratadas; identificándose VPH-16 en 3/6 de los casos positivos. En Paraguay existe alta incidencia de lesiones pre-neoplásicas y CCU siendo un problema de salud pública. Los datos observados sugieren que la utilización de este sistema para la detección viral puede llevar a optimizar el seguimiento post-tratamiento y la identificación de VPH-16 y 18 podría contribuir a la selección de pacientes en mayor riesgo de desarrollar una lesión cervical que deben someterse a una vigilancia frecuente y meticulosa.

Cervical cancer is the leading cause of cancer death in women in developing country.Persistent infection with human papillomavirus (HPV) is a necessary factor in premalignantlesions and cervical cancer. The Pap smear is the method most commonly used to detect thecervical cancer and its combined with the detection of viral DNA six months post-treatment increases effectiveness to identify women treated in risk with residual/recurrent lesion. Theaim was to describe the frequency of high-risk HPV (HR) in treated women for squamousintraepithelial lesion (SIL) who attended the Cervical Pathology Service of Hospital San Pablofrom January to December/2014. It is cross sectional, descriptive study which included 80patients who attended the service for post-treatment control. The 4800 Cobas HPV Test(Roche) was used, which detects HPV-16 and 18, and a pool of 10 HR-HPV(31,33,35,39,45,51,52,56,58,59) and two "probable" high risk (66,68). Viral infection wasfound in 7.5% (6/80) of patients treated; identifying HPV-16 in 3/6 of positive cases. InParaguay there is a high incidence of pre-neoplastic lesions and cervical cáncer remains apublic health problem. The observed data suggest that the use of this system for viraldetection can lead to optimize the post-treatment monitoring and the identification of HPV-16and 18 could contribute to the selection of patients at increased risk of developing cervicalinjury should undergo to frequent and careful monitoring.

Serorreactividad al antígeno de superficie del virus de la hepatitis B en mujeres que acudieron al laboratorio central de salud pública. Asunción, Paraguay / Seroreactivity against the hepatitis B virus surface antigen in women who attended the central public health laboratory. Asunción, Paraguay

La hepatitis B es un grave problema de salud pública a nivel mundial, aproximadamente cerca de 2 billones de personas tienen evidencia serológica de infección por el virus de la hepatitis B. El objetivo de este trabajo fue describir la frecuencia de hepatitis B e identificar los factores de riesgo asociados en mujeres en edad fértil que acudieron al Laboratorio Central de Salud Pública entre diciembre de 2013 y junio de 2014. Fue un estudio observacional analítico de corte transverso que, previo consentimiento informado, analizó suero de mujeres entre 15 y 44 años con una edad promedio de 26,6 (±6,8) años. Mediante la detección del antígeno de superficie de la hepatitis B por ELISA se identificaron seis casos positivos (0,4%), indicando una endemicidad baja; cifra que ha variado según perfil socio demográfico: según edad, las de 20 y más años presentaron una frecuencia mayor en comparación a las demás (p>0,05). No se observaron diferencias significativas al evaluar la seropositividad según el estado civil, el nivel de escolaridad, la condición de gravidez, los antecedentes de transfusiones, sin embargo, la seropositividad era mayor en las portadoras de tatuajes/piercing que entre las no portadoras, lo que representaba un riesgo 6,2 veces mayor (OR:6,2 IC95%:1,3-31,3). En conclusión, la frecuencia del HBsAg en nuestra población es baja, y el factor de riesgo asociado a su detección fue la presencia de tatuajes y/o piercing.

Hepatitis B is a serious public health problem worldwide; approximately about 2 billionpeople have serologic evidence of infection with hepatitis B virus. The aim of this analyticcross-sectional study was to describe the frequency of hepatitis B and identify risk factorsin women of child bearing age who attended the Central Public Health Laboratory in theperiod 2013 to 2014. Prior informed consent, antigen detection of hepatitis B surface wasperformed by ELISA in women between 15 and 44 years with a mean age of 26.6 (±6.8)years. The identification of six serologic positive cases (0.4%) indicates low endemicity.This figure varied according to socio-demographic profile: according to age, those whowere 20 years old or older had an increased frequency compared to the others (p> 0.05). No significant differences were observed in seropositivity by marital status, level ofeducation, pregnancy, history of transfusion, while seropositivity was higher amongcarriers of tattoos/piercing than among non-carriers, which represented a 6.2 times higherrisk (OR 6.2 95% CI 1.3 to 31.3). In conclusion, the frequency of HBsAg in our populationwas low. The risk factor associated with its detection was the presence of tattoos and / or piercings.

Detección de Metapneumovirus Humano en niños menores de 5 años hospitalizados en Paraguay / Detection of Human Metapneumovirus in Children under Age 5 Hospitalized in Paraguay

El Metapneumovirus humano (hMPV) constituye una importante causa de Infecciones Respiratorias Agudas (IRAs) en niños hospitalizados. En Paraguay, las IRAs están entre las primeras causas de hospitalización durante la infancia, siendo los virus los principales agentes causales. Sin embargo, aún persiste una alta proporción de casos sin etiología identificada. La pandemia de Influenza en el año 2009, condujo a una intensificación de la vigilancia de las infecciones respiratorias, lo cual permitió al mismo tiempo la búsqueda de otros virus como el hMPV. Nuestro objetivo fue detectar hMPV en niños hospitalizados por IRAs en Paraguay durante el año 2009. Fueron estudiadas 240 muestras respiratorias de niños < 5 años internados por IRAs en Paraguay durante el año 2009, que habían resultado negativas para otros virus respiratorios. Fue utilizado el reactivo LightMix Kit human MPV de TIBMOLBIOL, para la detección del gen N de hMPV por PCR en Tiempo Real, siguiendo el procedimiento indicado por el fabricante. De las 240 muestras estudiadas, 29 (12%) resultaron positivas para hMPV, con la mayor detección en julio y agosto; predominando en mayores de 1 año. Los principales signos y síntomas fueron tos, fiebre y dificultad respiratoria; y las complicaciones más frecuentes neumonía y bronquiolitis. Estos resultados proveen las primeras evidencias en Salud Pública, de la importancia del hMPV asociado a niños hospitalizados por IRAs en Paraguay.

Human metapneumovirus (hMPV) is a significant cause of acute respiratory infection (ARI) in hospitalized children. In Paraguay, ARI is one of the leading causes of childhood hospitalization, with viruses being the primary causal agents. However, a large number of cases of unknown etiology remain. The influenza pandemic of 2009 led to intensified vigilance concerning respiratory infections and more thorough efforts to confirm the presence of viruses such as hMPV. Our objective was to detect hMPV in children hospitalized for ARI in Paraguay during 2009. We studied respiratory samples from 240 children age < 5 years hospitalized for IRAs in Paraguay during 2009 who had tested negative for other respiratory viruses. We used the hMPV-reactive LightMix® kit from TIB MOLBIOL for the detection of the nucleoprotein (N) gene by real-time PCR according to manufacturer-specified procedures. Of the 240 samples studied, 29 (12%) were positive for hMPV, with the highest rates detected in July and August (winter) predominating in children over 1 year of age. The most common signs and symptoms were cough, fever, and respiratory distress; while the most common complications were pneumonia and bronchiolitis. These results provide the first evidence concerning the prevalence of hMPV in children hospitalized for ARI in Paraguay.

Phylogenetic relationship of dengue virus type 3 isolated in Brazil and Paraguay and global evolutionary divergence dynamics.

BACKGROUND: Dengue is the most important mosquito-borne viral disease worldwide. Dengue virus comprises four antigenically related viruses named dengue virus type 1 to 4 (DENV1-4). DENV-3 was re-introduced into the Americas in 1994 causing outbreaks in Nicaragua and Panama. DENV-3 was introduced in Brazil in 2000 and then spread to most of the Brazilian States, reaching the neighboring country, Paraguay in 2002. In this study, we have analyzed the phylogenetic relationship of DENV-3 isolated in Brazil and Paraguay with viruses isolated worldwide. We have also analyzed the evolutionary divergence dynamics of DENV-3 viruses. RESULTS: The entire open reading frame (ORF) of thirteen DENV-3 isolated in Brazil (n = 9) and Paraguay (n = 4) were sequenced for phylogenetic analysis. DENV-3 grouped into three main genotypes (I, II and III). Several internal clades were found within each genotype that we called lineage and sub-lineage. Viruses included in this study belong to genotype III and grouped together with viruses isolated in the Americas within the lineage III. The Brazilian viruses were further segregated into two different sub-lineage, A and B, and the Paraguayan into the sub-lineage B. All three genotypes showed internal grouping. The nucleotide divergence was in average 6.7% for genotypes, 2.7% for lineages and 1.5% for sub-lineages. Phylogenetic trees constructed with any of the protein gene sequences showed the same segregation of the DENV-3 in three genotypes. CONCLUSION: Our results showed that two groups of DENV-3 genotypes III circulated in Brazil during 2002-2009, suggesting different events of introduction of the virus through different regions of the country. In Paraguay, only one group DENV-3 genotype III is circulating that is very closely related to the Brazilian viruses of sub-lineage B. Different degree of grouping can be observed for DENV-3 and each group showed a characteristic evolutionary divergence. Finally, we have observed that any protein gene sequence can be used to identify the virus genotype.

Detección de metapneumovirus humano en niños menores de 5 años hospitalizados en Paraguay / Detection of human metapneumovirus in children under age 5 Hospitalized in Paraguay

El Metapneumovirus humano (hMPV) constituye una importante causa de Infecciones Respiratorias Agudas (IRAs) en niños hospitalizados. En Paraguay, las IRAs están entre las primeras causas de hospitalización durante la infancia, siendo los virus los principales agentes causales. Sin embargo, aún persiste una alta proporción de casos sin etiología identificada. La pandemia de Influenza en el año 2009, condujo a una intensificación de la vigilancia de las infecciones respiratorias, lo cual permitió al mismo tiempo la búsqueda de otros virus como el hMPV. Nuestro objetivo fue detectar hMPV en niños hospitalizados por IRAs en Paraguay durante el año 2009. Fueron estudiadas 240 muestras respiratorias de niños <5 años internados por IRAs en Paraguay durante el año 2009, que habían resultado negativas para otros virus respiratorios. Fue utilizado el reactivo LightMix Kit human MPV de TIBMOLBIOL, para la detección del gen N de hMPV por PCR en Tiempo Real, siguiendo el procedimiento indicado por el fabricante. De las 240 muestras estudiadas, 29 (12%) resultaron positivas para hMPV, con la mayor detección en julio y agosto; predominando en mayores de 1 año. Los principales signos y síntomas fueron tos, fiebre y dificultad respiratoria; y las complicaciones más frecuentes neumonía y bronquiolitis. Estos resultados proveen las primeras evidencias en Salud Pública, de la importancia del hMPV asociado a niños hospitalizados por IRAs en Paraguay.