Development and Application of An Assay for High-throughput Antiviral Compounds Screening against Alphaviruses / 病毒学报
Chinese Journal of Virology
;
(6): 641-646, 2015.
Artigo
em Chinês
| WPRIM
| ID: wpr-296235
ABSTRACT
To establish a cell-based rapid luciferase suppression assay for high-throughput screening (HTS) anti-alphaviruses compounds screening, which could cause viral encephalitis, raise the social issues associated directly with public health and huge economic burden to the society. The Gaussia luciferase assay system was used for HTS model for identifying inhibitors of labeled virus XJ160-GLUC. The decreased 50% GLUC activity inhibition ratio was deemed to be the screening positive index. The reaction system in this model was optimized, and the reliability of the model was evaluated. For HTS model's optimization, cells were infected with XJ160-GLUC at an MOI of 0.025 PFU/cell. The supernatant treated with compounds 48h were collected for GLUC expression detection. In the model, Z' factor was up to 0.71, demonstrating that HTS assay for identifying inhibitors that target all aspects of the viral life cycle of XJ160-GLUC was stable and reliable. After screening 8080 compounds (five-in-one), 341 positive samples were selected, and the positive rate was 4.2% with a cutoff at 50% inhibition. Then 1705 compounds were screened subsequently and the positive rate was 1.1% with obtaining 19 positive compounds. These results will lay the foundation for finding the anti-alphaviruses' drug targets.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Antivirais
/
Farmacologia
/
Genes Reporter
/
Alphavirus
/
Avaliação Pré-Clínica de Medicamentos
/
Ensaios de Triagem em Larga Escala
/
Genética
/
Luciferases
/
Metabolismo
/
Métodos
Tipo de estudo:
Estudo diagnóstico
/
Estudo prognóstico
/
Estudo de rastreamento
Limite:
Animais
Idioma:
Chinês
Revista:
Chinese Journal of Virology
Ano de publicação:
2015
Tipo de documento:
Artigo
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