Combining bacteriophage engineering and linear dichroism spectroscopy to produce a DNA hybridisation assay.
Combining bacteriophage engineering and linear dichroism spectroscopy to produce a DNA hybridisation assay.
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DOI：
10.1039/d0cb00135j
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发表时间：
2020-12-01
影响因子：
4.1
通讯作者：
Tucker JHR
中科院分区：
文献类型：
--
作者：
Ali A;Little HA;Carter JG;Douglas C;Hicks MR;Kenyon DM;Lacomme C;Logan RT;Dafforn TR;Tucker JHR
核酸检测是我们生物检测库的重要组成部分，COVID-19大流行清楚地表明了快速高效检测特定致病序列对医疗保健的重要性。作为建立新的DNA检测方法和信号读出的一部分，我们在这里展示了线性二色（LD）光谱如何用于产生快速和模块化的检测系统，用于检测来自细菌和病毒病原体的DNA数量。LD传感方法利用DNA位点共价附着在噬菌体M13表面的生物纳米颗粒（噬菌体M13）的流体排列变化，通过DNA靶点和两个M13生物纳米颗粒之间形成互补双链诱导读出信号。这种新的三明治检测方法可以在1分钟内检测到低至皮摩尔水平的致病性物质，而无需扩增，成功检测到植物病毒（马铃薯病毒Y）和氨苄西林抗性基因ampR的DNA序列就证明了这一点。本文描述了一种基于LD光谱和生物纳米颗粒支架的新型DNA传感方法，该方法可以快速检测与细菌和病毒病原体相关的DNA链。
Nucleic acid detection is an important part of our bio-detection arsenal, with the COVID-19 pandemic clearly demonstrating the importance to healthcare of rapid and efficient detection of specific pathogenic sequences. As part of the drive to establish new DNA detection methodologies and signal read-outs, here we show how linear dichroism (LD) spectroscopy can be used to produce a rapid and modular detection system for detecting quantities of DNA from both bacterial and viral pathogens. The LD sensing method exploits changes in fluid alignment of bionanoparticles (bacteriophage M13) engineered with DNA stands covalently attached to their surfaces, with the read-out signal induced by the formation of complementary duplexes between DNA targets and two M13 bionanoparticles. This new sandwich assay can detect pathogenic material down to picomolar levels in under 1 minute without amplification, as demonstrated by the successful sensing of DNA sequences from a plant virus (Potato virus Y) and an ampicillin resistance gene, ampR. A novel DNA sensing method based on LD spectroscopy and using bionanoparticle scaffolds is described, as demonstrated by the rapid detection of DNA strands associated with bacterial and viral pathogens.