Division Biophotonics , Federal Institute for Materials Research and Testing (BAM) , Richard-Willstaetter Strasse 11 , 12489 , Berlin , Germany.
Department of Chemistry , Humboldt University of Berlin , Brook-Taylor-Strasse 2 , 12489 Berlin , Germany.
Bioconjug Chem. 2018 May 16;29(5):1690-1702. doi: 10.1021/acs.bioconjchem.8b00157. Epub 2018 May 3.
Detection of pathogenic nucleic acids remains one of the most reliable approaches for the diagnosis of a broad range of diseases. Current PCR-based methods require experienced personnel and cannot be easily used for point-of-care diagnostics, making alternative strategies for the sensitive, reliable, and cost-efficient detection of pathogenic nucleic acids highly desirable. Here, we report an enzyme-free method for the fluorometric detection of RNA that relies on a target-induced fluorophore transfer onto a semiconductor quantum dot (QD), uses PNA probes as selective recognition elements and can be read out with simple and inexpensive equipment. For QD-PNA conjugates with optimized PNA content, limits of detection of dengue RNA in the range of 10 pM to 100 nM can be realized within 5 h in the presence of a high excess of noncomplementary RNA.
检测致病核酸仍然是诊断广泛疾病最可靠的方法之一。目前基于 PCR 的方法需要有经验的人员，并且不能轻易用于即时诊断，因此非常需要替代策略来灵敏、可靠且经济高效地检测致病核酸。在这里，我们报告了一种无酶方法，用于依赖于靶标诱导的荧光团转移到半导体量子点 (QD) 上的 RNA 的荧光检测，使用 PNA 探针作为选择性识别元件，并且可以使用简单且廉价的设备进行读取。对于具有优化 PNA 含量的 QD-PNA 缀合物，可以在存在大量非互补 RNA 的情况下，在 5 小时内实现登革热 RNA 的检测限在 10 pM 到 100 nM 的范围内。
