Application of QuantiGene (TM) nucleic acid quantification technology for high throughput screening

To identify inhibitors of interleukin-8 (IL-8) production, a high throughpu t assay was developed using the QuantiGene(TM) nucleic acid quantification kit that employs branched-chain DNA (bDNA) technology to measure the mRNA d irectly from cells. Unlike polymerase chain reaction and other technologies that employ target amplification, the QuantiGene system uses signal amplif ication. To perform the assay, various molecular probes capable of hybridiz ing with IL-8 mRNA were designed and synthesized. A human lung epithelial c ell line was treated with interleukin-1 alpha (IL-1 alpha) to stimulate the IL-8 gene expression and the mRNA was measured using the QuantiGene system . The QuantiGene assay was sensitive, flexible, and reproducible and achiev ed equivalent or better sensitivity than promoter-reporter assays, and elim inated the time required for constructing a promoter-reporter system. Our d ata show that bDNA technology has the potential to be used as a high throug hput screening assay.