Epidemiología molecular y análisis filogenético de la infección por el virus del papiloma humano en mujeres con lesiones cervicales y cáncer en la región litoral del Ecuador

The aim of the present study was to gather information regarding the molecular epidemiology of Human papillomavirus (HPV) and related risk factors in a group of women with low- and high-grade cervical lesions and cancer from the coastal region of Ecuador. In addition, we studied the evolution of HPV variants from the most prevalent types and provided a temporal framework for their emergence, which may help to trace the source of dissemination within the region. We analyzed 166 samples, including 57 CIN1, 95 CIN2/3 and 14 cancer cases. HPV detection and typing was done by PCR-sequencing (MY09/MY11). HPV variants and estimation of the time to most recent common ancestor (tMRCA) was assessed through phylogeny and coalescence analysis. HPV DNA was found in 54.4% of CIN1, 74.7% of CIN2/3 and 78.6% of cancer samples. HPV16 (38.9%) and HPV58 (19.5%) were the most prevalent types. Risk factors for the development of cervical lesions/cancer were the following: three or more pregnancies (OR = 4.3), HPV infection (OR = 3.7 for high-risk types; OR = 3.5 for HPV16), among others. With regard to HPV evolution, HPV16 isolates belonged to lineages A (69%) and D (31%) whereas HPV58 isolates belonged only to lineage A. The period of emergence of HPV16 was in association with human populations (tMRCA = 91. 052 years for HPV16A and 27. 000 years for HPV16D), whereas HPV58A preceded Homo sapiens evolution (322. 257 years). This study provides novel data on HPV epidemiology and evolution in Ecuador, which will be fundamental in the vaccine era.Fil: Bedoya Pilozo, Cesar H.. Escuela Superior Politécnica del Litoral; Ecuador. Instituto Nacional de Investigaciones en Salud Pública; EcuadorFil: Medina Magües, Lex G.. Escuela Superior Politécnica del Litoral; EcuadorFil: Espinosa García, Maylen. Instituto Nacional de Investigaciones en Salud Pública; EcuadorFil: Sánchez, Martha. Instituto Nacional de Investigaciones en Salud Pública; EcuadorFil: Parrales Valdiviezo, Johanna V.. Instituto Nacional de Investigaciones en Salud Pública; EcuadorFil: Molina, Denisse. Instituto Nacional de Investigaciones en Salud Pública; EcuadorFil: Ibarra, María A.. Instituto Nacional de Investigaciones en Salud Pública; EcuadorFil: Quimis Ponce, María. Instituto Nacional de Investigaciones en Salud Pública; EcuadorFil: España, Karool. Instituto Nacional de Investigaciones en Salud Pública; EcuadorFil: Párraga Macias, Karla E.. Instituto Nacional de Investigaciones en Salud Pública; EcuadorFil: Cajas Flores, Nancy V.. Instituto Nacional de Investigaciones en Salud Pública; EcuadorFil: Solon, Orlando A.. Instituto Nacional de Investigaciones en Salud Pública; Ecuador. Universidad Agraria del Ecuador; EcuadorFil: Robalino Penaherrera, Jorge A.. Instituto Nacional de Investigaciones en Salud Pública; EcuadorFil: Chedraui, Peter. Hospital Gineco-Obstétrico Enrique C. Sotomayor; EcuadorFil: Escobar, Saul. Universidad Católica de Guayaquil; EcuadorFil: Loja Chango, Rita D.. Universidad Católica de Guayaquil; EcuadorFil: Ramirez Morán, Cecibel. Universidad Católica de Guayaquil; EcuadorFil: Espinoza Caicedo, Jasson. Universidad Católica de Guayaquil; EcuadorFil: Sánchez Giler, Sunny. Universidad Especialidades Espíritu Santo. Facultad de Ciencias Médicas; EcuadorFil: Limia, Celia M.. Instituto de Medicina Tropical Pedro Kouri; CubaFil: Alemán, Yoan. Instituto de Medicina Tropical Pedro Kouri; CubaFil: Soto, Yudira. Instituto de Medicina Tropical Pedro Kouri; CubaFil: Kouri, Vivian. Instituto de Medicina Tropical Pedro Kouri; CubaFil: Culasso, Andrés Carlos Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Virología; ArgentinaFil: Badano, Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Secretaría de Educación Superior, Ciencia, Tecnología e Innovación; Ecuador. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales. Laboratorio de Biología Molecular Aplicada; Argentin

Detection of Type 16 Human Papillomavirus DNA in Formalin-fixed Invasive Squamous Cells from Laryngeal Cancers by Polymerase Chain Reaction

To date approximately 77 distinct human papillomavirus (HPV) genotypes have been described and the genomic sequences of most of them have been fully analyzed. About 30 additional partial sequences have been obtained from putative novel HPV types, suggesting that the total number of existing HPV genotypes is in excess of 100. HPV cause benign tumors in the respiratory tract. Mounting evidence suggests that they also play a role in the etiology of a subset of head and neck cancers. Carcinomas in patients with a history of recurrent respiratory papillomatosis are caused clearly by persistent HPV interacting with one or more carcinogenic agents. Previous studies have suggested squamous neoplasia of the larynx and may also be HPV related

Prevalence of Chlamydia trachomatis in Human Immunodeficiency Virus-infected Women in Cuba

To determine the prevalence rates and serovar distribution of Chlamydia trachomatis   cervical infections in Cuban women, two different groups were selected. Group I consisted of 60 human immunodeficiency virus (HIV-1) seropositive women from different regions of Cuba and group II of 60 randomly selected women HIV seronegative and apparently healthy. C. trachomatis was detected in cervical scrapes by mean of nested polymerase chain reaction (PCR) specific for major out membrane protein. The overall prevalence rate of C. trachomatis in cervical scrapes determined by nested PCR was 10% in group I and the estimated prevalence was 6.6% for group II; 83.3% of HIV seropositive women with C. trachomatis infection reported history of pelvic inflammatory disease followed by cervicitis (50%). The control group C. trachomatis-infected women referred a history of cervicitis in 75% of cases. Other reports in the latter group included infertility and pelvic inflamatory disease in 50%. The present study is the first report of C. trachomatis prevalence in Cuba. It showed that there was not significantly difference in the prevalence rate of C. trachomatis between both groups

Analysis of respiratory syncytial virus in clinical samples by reverse transcriptase-polymerase chain reaction restriction mapping

The aim of this study was to develop a polymerase chain reaction (PCR) for the detection of respiratory syncytial virus (RSV) genomes. The primers were designed from published sequences and selected from conserved regions of the genome encoding for the N protein of subgroups A and B of RSV. PCR was applied to 20 specimens from children admitted to the respiratory ward of "William Soler" Pediatric Hospital in Havana City with a clinical diagnosis of bronchiolitis. The PCR was compared with viral isolation and with an indirect immunofluorescence technique that employs monoclonal antibodies of subgroups A and B. Of 20 nasopharyngeal exudates, 10 were found positive by the three assayed methods. In only two cases, samples that yielded positive RNA-PCR were found negative by indirect immunofluorescence and cell culture. Considering viral isolation as the "gold standard" technique, RNA-PCR had 100% sensitivity and 80% specificity. RNA-PCR is a specific and sensitive technique for the detection of the RSV genome. Technical advantages are discussed

Detección y tipificación de papilomavirus humano en lesiones condilomatosas anogenitales de hombres cubanos seropositivos al VIH-1

Introduction. In the general HIV-infected population, there are few data on the prevalence and risk factors for anogenital condyloma, precursor lesions for anogenital cancer. In Cuba, there have been few studies about the prevalence of HPV in HIV infected people. Objective. Detection and typing HPV in anogenital condyloma lesions from Cuban HIV-1 infected men. Materials and Methods. We studied samples of warts from 30 Cuban patients attending the Instituto Pedro Kouri. HPV DNA was detected using PCR with MY09/11 primers. HPV DNA typing was conducted by hybridization with 32 low and high-risk HPV DNA biotin-labelled probes. Clinic, epidemiologic and immunologic features of these patients were analyzed. Results. HPV DNA was detected in 100% of the lesions sampled. Associated with the condyloma lesions, HPV 6 and 11 were detected in 19 of 30 cases (63.3%) and 16 of 30 patients (53.3%), respectively. Multiple infections with high-risk HPV types, predominantly HPV 16 (50%) and 18 (43.3%) were observed in 19 samples, with up to 11 different genotypes in the same patient. Condylomas were frequently localized in perianal region.Trichloroacetic acid (80%) was the most used therapy. In patients with a T lymphocyte CD4+ count < 500 cell/mm3 high-risk HPV types were detected more frequently. This correlation was statistically significant [X2 =11.27; RR=28.5; 95% IC: 2.6-306.6 (p=0.008)]. Conclusions. High and low risk HPV types detection in condylomas from HIV-infected men and T lymphocyte CD4 count should be considered for the monitoring and early detection of anogenital neoplasia. These findings could be a valuable data for follow-up and treatments.Introducción. Los condilomas son causados por Papilomavirus Humano (PVH), esencialmente tipos 6 y 11. No se conoce la prevalencia de PVH en hombres cubanos seropositivos al Virus de la Inmunodeficiencia Humana (VIH). Objetivo. Detectar infección por PVH, genotipos y aspectos clínicos en hombres cubanos seropositivos al VIH-1. Materiales y Métodos. Se estudiaron muestras de condilomas en 30 pacientes cubanos atendidos en el Instituto �Pedro Kourí�. Al tejido se le realizó análisis histopatológico y detección de PVH mediante Reacción en Cadena de la Polimerasa (RCP) con oligos MY09/11. Además, genotipificación con 32 sondas biotinadas, para PVH de alto o bajo riesgo oncogénico. Resultados. El ADN de PVH fue detectado en 100% de las muestras. Los genotipos 6 y 11 se identificaron en 63.3% y en 53.3% de los casos, respectivamente. En 19 individuos, se demostró coinfección con genotipos de bajo y alto riesgo. En un mismo paciente se pudieron detectar hasta 11 genotipos, con predominio de PVH 16 (50%) y 18 (43.3%). La terapia más usada fue el ácido tricloracético. Se observó asociación estadística [X2 =11.27; RR=28.5; 95% IC: 2.6- 306.6 (p=0.008)] entre la presencia de genotipos de alto riesgo oncogénico y el conteo de linfocitos T CD4+ < 500 células /mm3. Conclusiones. La detección de PVH de alto y bajo riesgo oncogénico, en lesiones condilomatosas de pacientes cubanos seropositivos al VIH-1, en relación con el conteo de CD4+ es un hallazgo de interés para el seguimiento y la detección temprana de neoplasias anogenitales