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J Virol Methods ; 179(1): 242-9, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22115788

RESUMO

Rotaviruses are an important cause of severe gastroenteritis in children under two years of age. Two vaccines have become recently available, however, there are no specific pharmacological interventions of rotavirus disease. Recently, libraries of siRNAs or libraries of chemical compounds that can be tested for their ability to inhibit biological processes have been developed. To search these libraries for drugs or siRNAs that may prevent rotavirus replication it is necessary to have methods for high-throughput screening. In this study several methods to quantify rotavirus replication in cell culture were evaluated; the cell death and viral protein expression assays were compared, and an in-cell Western method based on infrared detection that allows the simultaneous quantification of viral antigen and total protein content in the same cell culture well was developed. This is an easy, inexpensive method for detection of viral replication, and it is compatible with high-throughput screening.


Assuntos
Antivirais/isolamento & purificação , Produtos Biológicos/isolamento & purificação , Avaliação Pré-Clínica de Medicamentos/métodos , Ensaios de Triagem em Larga Escala/métodos , RNA Interferente Pequeno/isolamento & purificação , Rotavirus/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos , Animais , Antígenos Virais/biossíntese , Sobrevivência Celular , Células Cultivadas , Humanos
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