Fluorogenic DNAzyme Probes as Bacterial Indicators
Fluorogenic DNAzyme Probes as Bacterial Indicators
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DOI：
10.1002/anie.201100477
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发表时间：
2011-01-01
影响因子：
16.6
通讯作者：
Li, Yingfu
中科院分区：
文献类型：
--
作者：
Ali, M. Monsur;Aguirre, Sergio D.;Li, Yingfu
食源性病原体的流行、抗药性细菌和病毒的出现以及生物恐怖主义的威胁是我们这个时代最紧迫的问题。病原体的早期检测是防止大规模疫情爆发的关键步骤，在商业全球化和旅行时间缩短显著增加了传染源传播的速度和范围的今天尤为重要。病原体检测传统上是通过使用微生物学技术进行的，这种技术非常准确，但可能需要几天（甚至几周）才能得到结果。[1]基于抗体和PCR的检测都大大缩短了检测时间;然而，这些检测仍然需要多个步骤和专用设备。[2]对于可以实现已知病原体的快速检测的简单方法和可以快速到位以在意外爆发中创建针对新病原体的测定的新平台，存在显著的需求。这些考虑促使我们开发一种基于催化DNA分子（DNA酶）的平台，DNA酶是一类特殊的功能核酸[3]，是具有催化能力的人工单链DNA分子。[4]这些分子可以通过体外选择从随机序列DNA库中分离[5]，并且已经越来越多地被探索作为各种应用的分子工具。[4，6]在本文中，我们证明了可以从DNA文库中分离荧光DNA酶，以在由特定细菌病原体产生的粗细胞外混合物（CEM）中发荧光，并且这种探针可以用于开发简单的“混合和读取”测定以检测该病原体。
The prevalence of food-borne pathogens, emergence of drugresistant bacteria and viruses, and threat of bioterrorism are amongst the most pressing concerns of our time. The early detection of pathogens is a crucial step in preventing largescale outbreaks, and is particularly important today as globalization of commerce and shorter journey times have significantly increased the rate and breadth of the spread of infectious agents.Pathogen detection is traditionally performed by using microbiological techniques, which are highly accurate but can take days (even weeks) to obtain a result.[1] Both antibodyand PCR-based tests offer much-reduced detection times; however these tests still require multiple steps and specialized equipment.[2] There is a significant need for both simple methods that can achieve rapid detection of known pathogens and for new platforms that can be quickly put in place to create assays for a new pathogen in an unanticipated outbreak. These considerations have motivated us to develop a platform based on catalytic DNA molecules (DNAzymes), which are a special class of functional nucleic acids [3] that are artificial single-stranded DNA molecules that have a catalytic ability.[4] These molecules can be isolated from a randomsequence DNA pool by in vitro selection [5] and have been increasingly explored as molecular tools for various applications.[4, 6] Herein we demonstrate that fluorogenic DNAzymes can be isolated from a DNA library to fluoresce in the crude extracellular mixture (CEM) that is produced by a specific bacterial pathogen, and that such probes can be used to develop a simple “mix-and-read” assay to detect this pathogen.