[Identification of adenovirus associated with conjunctivitis by molecular methodology]. / Identificación por métodos moleculares de adenovirus asociados a conjuntivitis.

OBJECTIVE: To identify adenovirus in patients with conjunctivitis by molecular methods such as the Polymerase Chain Reaction (PCR) and DNA sequencing. METHODS: Samples of scrapings from the inferior fornix of 51 patients (39 diagnosed with Follicular Conjunctivitis and 12 diagnosed with Vernal Conjunctivitis) were processed by generic PCR to identify adenovirus. All the samples that were PCR positive were cultured on VERO cells for virus isolation, with this being demonstrated by immunofluorescence. For the identification of the isolated serotype, the multiplex PCR was utilized and DNA automated sequencing was employed to identify the genetic variants. RESULTS: Twenty-eight of the individuals diagnosed with Follicular Conjunctivitis and six of those diagnosed with Vernal Conjunctivitis, had positive results to adenovirus (67%). The cultures in VERO cells allowed the isolation of eight samples. Only three of the isolated viruses (one Ad1 and two Ad2) were identified by the multiplex PCR used to identify the subgenus C adenovirus. An Ad1 genetic variant was identified by automated sequencing while the Ad2 serotypes were identical to the ones reported by Genbank. CONCLUSIONS: The Polymerase Chain Reaction and DNA sequencing are useful tools to identify and characterize microorganisms responsible for diseases such as conjunctivitis caused by adenoviruses.

Identificación por métodos moleculares de adenovirus asociados a conjuntivitis / Identification of adenovirus associated with conjunctivitis by molecular methodology

Objetivo: Por medio de métodos moleculares comola Reacción en Cadena de la Polimerasa (PCR) y lasecuenciación del material genético, identificar adenovirusen pacientes con conjuntivitis.Métodos: Se procesaron muestras de raspado delsaco conjuntival inferior de 51 pacientes (39 diagnosticadoscon conjuntivitis folicular y 12 diagnosticadoscon conjuntivitis vernal) para identificaradenovirus por medio de la PCR genérica. Todas lasmuestras positivas en la PCR genérica, fueron cultivadasen células VERO para el aislamiento delvirus, éste se demostró por inmunofluorescencia. Seutilizó la PCR múltiple para la caracterización delos serotipos aislados y las variantes genéticas seidentificaron mediante la secuenciación automatizada.Resultados: Veintiocho de los pacientes diagnosticadoscon conjuntivitis folicular y seis de los diagnosticadoscon conjuntivitis vernal resultaron positivosa adenovirus (67%). El cultivo en célulasVERO permitió el aislamiento del virus de 8 muestras.Sólo tres de los aislados fueron identificados(un serotipo Ad1 y dos Ad2) por la PCR múltipleque identifica adenovirus del subgénero C. Por estreptomicisecuenciaciónautomatizada se identificó unavariante genética correspondiente al Ad1, mientrasque los aislados de Ad2 fueron idénticos a losreportados en el Banco de Genes.Conclusiones: La Reacción en Cadena de la Polimerasay la Secuenciación son herramientas útilespara la identificación y caracterización de agentescausantes de enfermedades tales como la Conjuntivitispor Adenovirus

Objective: To identify adenovirus in patients with conjunctivitis by molecular methods such as the Polymerase Chain Reaction (PCR) and DNA sequencing. Methods: Samples of scrapings from the inferior fornix of 51 patients (39 diagnosed with Follicular Conjunctivitis and 12 diagnosed with Vernal Conjunctivitis) were processed by generic PCR to identify adenovirus. All the samples that were PCR positive were cultured on VERO cells for virus isolation, with this being demonstrated by immunofluorescence. For the identification of the isolated serotype, the multiplex PCR was utilized and DNA automated sequencing was employed to identify the genetic variants. Results: Twenty-eight of the individuals diagnosed with Follicular Conjunctivitis and six of those diagnosed with Vernal Conjunctivitis, had positive results to adenovirus (67%). The cultures in VERO cells allowed the isolation of eight samples. Only three of the isolated viruses (one Ad1 and two Ad2) were identified by the multiplex PCR used to identify the subgenus C adenovirus. An Ad1 genetic variant was identified by automated sequencing while the Ad2 serotypes were identical to the ones reported by Genbank. Conclusions: The Polymerase Chain Reaction and DNA sequencing are useful tools to identify and characterize microorganisms responsible for diseases such as conjunctivitis caused by adenoviruses