SERS primers and their mode of action for pathogen DNA detection.
SERS primers and their mode of action for pathogen DNA detection.
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DOI：
10.1021/ac302254h
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发表时间：
2013-02
影响因子：
7.4
通讯作者：
D. van Lierop;I. A. Larmour;K. Faulds;D. Graham
中科院分区：
文献类型：
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作者：
D. van Lierop;I. A. Larmour;K. Faulds;D. Graham
Sers引物已被用于直接检测特异性PCR产物，利用单链和双链DNA在纳米颗粒表面上的吸附差异。在此，七个参数重要的改进积极Sers测定真实的应用进行了研究。首先，我们应用模型系统进行优化实验，然后通过PCR检测病原体DNA，然后引入一种新的检测方法，该方法利用Taq DNA聚合酶的5 '→ 3'外切核酸酶活性来部分消化Sers探针，产生染料标记的单链DNA，从而增加用于检测病原体DNA的Sers信号。应用该模型体系，发现单分子Sers引物比双分子Sers引物表现得更好。然而，在PCR测定中，发现单分子和双分子Sers引物的表现非常相似，并且使用5 '→ 3'核酸外切酶消化测定获得了最可重复的结果。这些基于SERS的检测方法提供了优于传统荧光技术的新途径，而不会影响灵敏度或选择性。
SERS primers have been used to directly detect specific PCR products utilizing the difference in adsorption of single-stranded and double-stranded DNA onto nanoparticle surfaces. Herein, seven parameters important for improved positive SERS assays for real applications were investigated. First, we applied a model system for optimization experiments, followed by a PCR assay to detect pathogen DNA, and then the introduction of a new assay that utilizes the 5'→3' exonuclease activity of Taq DNA polymerase to partly digest the SERS probe, generating dye-labeled single-stranded DNA increasing the SERS signals for detection of pathogen DNA. Applying the model system, it was found that uni-molecular SERS primers perform better than bi-molecular SERS primers. However, within the PCR assays, it was found that uni- and bi-molecular SERS primers performed very similarly, and the most reproducible results were obtained using the 5'→3' exonuclease digestion assay. These SERS-based assays offer new routes over conventional fluorescence-based techniques without compromising sensitivity or selectivity.