Development and Application of An Assay for High-throughput Antiviral Compounds Screening against Alphaviruses.
- Author:
Yan WEI
;
Jianjiao LI
;
Huanqin WANG
;
Shan CEN
;
Guodong LIANG
;
Wenjie TAN
;
Wuyang ZHU
- Publication Type:Journal Article
- MeSH:
Alphavirus;
drug effects;
genetics;
metabolism;
Animals;
Antiviral Agents;
pharmacology;
Drug Evaluation, Preclinical;
methods;
Genes, Reporter;
High-Throughput Screening Assays;
methods;
Luciferases;
genetics;
metabolism
- From:
Chinese Journal of Virology
2015;31(6):641-646
- CountryChina
- Language:Chinese
-
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.
