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一种用于检测特定 DNA 靶标的新型 SERRS 夹心杂交分析方法。

A novel SERRS sandwich-hybridization assay to detect specific DNA target.

机构信息

Laboratoire de Géologie de Lyon-Terre Planètes Environnement, ENS Lyon, Université Lyon 1, CNRS, Ecole Normale Supérieure de Lyon, Lyon, France.

出版信息

PLoS One. 2011;6(5):e17847. doi: 10.1371/journal.pone.0017847. Epub 2011 May 31.

Abstract

In this study, we have applied Surface Enhanced Resonance Raman Scattering (SERRS) technology to the specific detection of DNA. We present an innovative SERRS sandwich-hybridization assay that allows specific DNA detection without any enzymatic amplification, such as is the case with Polymerase Chain Reaction (PCR). In some substrates, such as ancient or processed remains, enzymatic amplification fails due to DNA alteration (degradation, chemical modification) or to the presence of inhibitors. Consequently, the development of a non-enzymatic method, allowing specific DNA detection, could avoid long, expensive and inconclusive amplification trials. Here, we report the proof of concept of a SERRS sandwich-hybridization assay that leads to the detection of a specific chamois DNA. This SERRS assay reveals its potential as a non-enzymatic alternative technology to DNA amplification methods (particularly the PCR method) with several applications for species detection. As the amount and type of damage highly depend on the preservation conditions, the present SERRS assay would enlarge the range of samples suitable for DNA analysis and ultimately would provide exciting new opportunities for the investigation of ancient DNA in the fields of evolutionary biology and molecular ecology, and of altered DNA in food frauds detection and forensics.

摘要

在本研究中,我们将表面增强共振拉曼散射(SERRS)技术应用于 DNA 的特异性检测。我们提出了一种创新的 SERRS 夹心杂交测定法,该方法无需任何酶扩增(如聚合酶链反应(PCR))即可进行特异性 DNA 检测。在某些基质中,如古老或加工后的残留物,由于 DNA 改变(降解、化学修饰)或抑制剂的存在,酶扩增会失败。因此,开发一种非酶方法,允许特异性 DNA 检测,可以避免冗长、昂贵和不确定的扩增试验。在这里,我们报告了一种 SERRS 夹心杂交测定法的概念验证,该方法可用于检测特定的羱羊 DNA。这种 SERRS 测定法显示出其作为 DNA 扩增方法(特别是 PCR 方法)的非酶替代技术的潜力,具有多种用于物种检测的应用。由于量和损伤类型高度依赖于保存条件,本 SERRS 测定法将扩大适合 DNA 分析的样品范围,并最终为进化生物学和分子生态学领域的古代 DNA 研究以及食品欺诈检测和法医学中的改变 DNA 研究提供令人兴奋的新机会。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fc8a/3104981/804d7d08f7d1/pone.0017847.g001.jpg

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