Warrior U, Fan Y, David C A, Wilkins J A, McKeegan E M, Kofron J L, Burns D J
Abbott Laboratories, Abbott Park, IL 60064, USA.
J Biomol Screen. 2000 Oct;5(5):343-52. doi: 10.1177/108705710000500506.
To identify inhibitors of interleukin-8 (IL-8) production, a high throughput assay was developed using the QuantiGene nucleic acid quantification kit that employs branched-chain DNA (bDNA) technology to measure the mRNA directly from cells. Unlike polymerase chain reaction and other technologies that employ target amplification, the QuantiGene system uses signal amplification. To perform the assay, various molecular probes capable of hybridizing with IL-8 mRNA were designed and synthesized. A human lung epithelial cell line was treated with interleukin-1alpha (IL-1alpha) 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 achieved equivalent or better sensitivity than promoter-reporter assays, and eliminated the time required for constructing a promoter-reporter system. Our data show that bDNA technology has the potential to be used as a high throughput screening assay.
为了鉴定白细胞介素-8（IL-8）产生的抑制剂，开发了一种高通量检测方法，该方法使用QuantiGene核酸定量试剂盒，该试剂盒采用分支链DNA（bDNA）技术直接测量细胞中的mRNA。与聚合酶链反应和其他采用靶标扩增的技术不同，QuantiGene系统使用信号放大。为了进行该检测，设计并合成了各种能够与IL-8 mRNA杂交的分子探针。用人白细胞介素-1α（IL-1α）处理人肺上皮细胞系以刺激IL-8基因表达，并使用QuantiGene系统测量mRNA。QuantiGene检测灵敏、灵活且可重复，其灵敏度与启动子报告基因检测相当或更高，并且无需构建启动子报告基因系统所需的时间。我们的数据表明，bDNA技术有潜力用作高通量筛选检测方法。
