• 文献标题:   A Fluorescence Assay for HIV-1 Ribonuclease H Activity and Inhibition Detection Using Graphene Oxide as Sensing Platform
  • 文献类型:   Article
  • 作  者:   LI X, DING XL, WANG GK, ZHOU YX, HOU RP, GAO SY, LU Y
  • 作者关键词:   graphene oxide, hiv1 rnase h activity detection, dnarna duplex
  • 出版物名称:   SCIENCE OF ADVANCED MATERIALS
  • ISSN:   1947-2935 EI 1947-2943
  • 通讯作者地址:   Henan Normal Univ
  • 被引频次:   7
  • DOI:   10.1166/sam.2014.1975
  • 出版年:   2014

▎ 摘  要

In this work, we have developed a simple and highly sensitive fluorescent biosensor for the detection of HIV-1 RNase H activity and inhibition by using graphene oxide (GO) as sensing platform. In our approach, a DNA-RNA substrate was prepared which a carboxy fluorescein (FAM) was labeled at the 3' termini of single strand DNA. The FAM labeled DNA-RNA substrate preserves most of the fluorescence when mixed with GO. However, in the present of RNase H, RNase H can cleavage the RNA into fragments, resulting in the dissociation of ssDNA and mononucleotides. The fluorescence intensity was greatly quenched after the addition of GO. Because of the super quenching ability of GO, the as-proposed method provides a wide linear range from 1.0 to 26.3 units mL(-1) and a low detection limit of 0.6 units mL(-1) for HIV-1 RNase H activity analysis. Additionally, this approach shows potential for high-throughput screening of HIV-1 RNase H inhibitors. The method not only provides a universal platform for monitoring activity and inhibition of RNase H but also process researches, drug discovery, and clinic diagnositcs.