RESUMEN
The recent increase in babies born with brain and eye malformations in Brazil is associated with Zika virus (ZIKV) infection in utero. ZIKV alters host DNA methylation in vitro. Using genome-wide DNA methylation profiling we compared 18 babies born with congenital ZIKV microcephaly with 20 controls. We found ZIKV-associated alteration of host methylation patterns, notably at RABGAP1L which is important in brain development, at viral host immunity genes MX1 and ISG15, and in an epigenetic module containing the causal microcephaly gene MCPH1. Our data support the hypothesis that clinical signs of congenital ZIKV are associated with changes in DNA methylation.
Asunto(s)
Metilación de ADN , Inmunidad/genética , Microcefalia/virología , Neurogénesis/genética , Infección por el Virus Zika , Encéfalo/crecimiento & desarrollo , Encéfalo/virología , Brasil , Proteínas de Ciclo Celular/genética , Preescolar , Proteínas del Citoesqueleto/genética , Femenino , Humanos , Lactante , Masculino , Embarazo , Complicaciones Infecciosas del Embarazo/virología , Virus Zika/inmunologíaRESUMEN
HPV infection is considered one of the leading causes of cervical cancer in the world. To date, more than 180 types of HPV have been described and viral typing is critical for defining the prognosis of cancer. In this work, a seminested PCR which allow fast and inexpensively detection and typing of HPV is presented. The system is based on the amplification of a variable length region within the viral gene E1, using three primers that potentially anneal in all HPV genomes. The amplicons produced in the first step can be identified by high resolution electrophoresis or direct sequencing. The seminested step includes nine specific primers which can be used in multiplex or individual reactions to discriminate the main types of HPV by amplicon size differentiation using agarose electrophoresis, reducing the time spent and cost per analysis.
Asunto(s)
Genoma Viral/genética , Papillomaviridae/clasificación , Infecciones por Papillomavirus/virología , Reacción en Cadena de la Polimerasa/métodos , Neoplasias del Cuello Uterino/virología , Proteínas Virales/genética , Secuencia de Aminoácidos , Cuello del Útero/virología , Cartilla de ADN/genética , Femenino , Genotipo , Técnicas de Genotipaje , Células HeLa , Humanos , Papillomaviridae/genética , Papillomaviridae/aislamiento & purificación , Sensibilidad y Especificidad , Alineación de SecuenciaRESUMEN
BACKGROUND: Surveillance is a critical component of any dengue prevention and control programme. Herein, we investigate the efficiency of the commercial kit Platelia Dengue NS1 Ag-ELISA to detect dengue virus (DENV) antigens in Aedes aegypti mosquitoes infected under laboratory conditions. METHODS: Under insectary conditions, four to five day-old mosquitoes were orally challenged with DENV-2 titer of 3.6 x 105 PFU equivalent/ml, incubated for 14 days and then killed. At ten time-points following mosquito death (0, 6, 12, 24, 72, 96, 120, 144 and 168 h), i.e., during a one-week period, dried mosquitoes were comparatively tested for the detection of the NS1 antigen with other methods of detection, such as qRT-PCR and virus isolation in C6/36 cells. RESULTS: We first observed that the NS1 antigen was more effective in detecting DENV-2 in Ae. aegypti between 12 and 72 h after mosquito death when compared with qRT-PCR. A second round involved comparing the sensitivity of detection of the NS1 antigen and virus isolation in C6/36 cells. The NS1 antigen was also more effective than virus isolation, detecting DENV-2 at all time-points, i.e., up to 168 h after mosquito death. Meanwhile, virus isolation was successful up to 96 h after Ae. aegypti death, but the number of positive samples per time period presented a tendency to decline progressively over time. From the 43 samples positive by the virus isolation technique, 38 (88.4%) were also positive by the NS1 test. CONCLUSION: Taken together, these results are the first to indicate that the NS1 antigen might be an interesting complementary tool to improve dengue surveillance through DENV detection in dried Ae. aegypti females.
Asunto(s)
Aedes/virología , Ensayo de Inmunoadsorción Enzimática/métodos , Proteínas no Estructurales Virales/aislamiento & purificación , Animales , Antígenos Virales , Línea Celular , Femenino , ARN Viral/aislamiento & purificación , Manejo de EspecímenesRESUMEN
We examined levels of dengue virus type 3 (DENV-3) RNA in association with the type of infection (primary or secondary) in 42 patients with fatal and non-fatal outcomes in Rio de Janeiro, Brazil, 2002. Subjects with fatal outcomes had mean virus titers significantly higher than those who survived (12.5 vs. 7.9 log(10) RNA copies/ml). Because primary infections were confirmed among the fatal cases (52.1%), antibody-dependent enhancement alone did not explain all the cases of severe disease in this study population. These findings suggest that high levels of DENV-3 may have contributed to the severe form of dengue in Rio de Janeiro, 2002.