Department of Physics and Astronomy, Texas Christian University, Fort Worth, Texas 76109, United States.
Center for Human Identification, University of North Texas Health Science Center, 3500 Camp Bowie Blvd., Fort Worth, Texas 76107, United States.
Anal Chem. 2022 Mar 29;94(12):5062-5068. doi: 10.1021/acs.analchem.1c05275. Epub 2022 Mar 14.
This article presents a novel approach to increase the detection sensitivity of trace amounts of DNA in a sample by employing Förster resonance energy transfer (FRET) between intercalating dyes. Two intercalators that present efficient FRET were used to enhance sensitivity and improve specificity in detecting minute amounts of DNA. Comparison of steady-state acceptor emission spectra with and without the donor allows for simple and specific detection of DNA (acceptor bound to DNA) down to 100 pg/μL. When utilizing as an acceptor a dye with a significantly longer lifetime (e.g., ethidium bromide bound to DNA), multipulse pumping and time-gated detection enable imaging/visualization of picograms of DNA present in a microliter of an unprocessed sample or DNA collected on a swab or other substrate materials.
本文提出了一种新方法，通过在嵌入染料之间发生Förster 共振能量转移（FRET），来提高样品中痕量 DNA 的检测灵敏度。使用两种呈现高效 FRET 的嵌入剂来提高灵敏度并改善检测微量 DNA 的特异性。通过比较有和没有供体的稳态受体发射光谱，可以简单而特异性地检测到低至 100pg/μL 的 DNA（与 DNA 结合的受体）。当使用具有显著更长寿命的染料（例如，与 DNA 结合的溴化乙锭）作为受体时，多脉冲泵浦和时间门控检测可实现对微升未经处理的样品中存在的皮克级 DNA 或在拭子或其他基底材料上收集的 DNA 的成像/可视化。
