Nonenzymatic detection of bacterial genomic DNA using the bio bar code assay
Nonenzymatic detection of bacterial genomic DNA using the bio bar code assay
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DOI：
10.1021/ac701626y
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发表时间：
2007-12-01
影响因子：
7.4
通讯作者：
Mirkin, Chad A.
中科院分区：
文献类型：
--
作者：
Hill, Haley D.;Vega, Rafael A.;Mirkin, Chad A.
据报道，通过基于非酶纳米材料的扩增方法（生物条形码测定）检测细菌基因组 DNA。该测定利用寡核苷酸功能化的磁性微粒从样品中捕获感兴趣的靶标。新测定中的一个关键步骤涉及在双链 DNA 热变性过程中使用封闭寡核苷酸。这些阻断剂在冷却后与目标 DNA 的特定区域结合，并防止双链 DNA 重新杂交，从而允许粒子探针结合。使用磁性颗粒分离目标后，寡核苷酸功能化的金纳米颗粒充当目标识别剂。纳米颗粒（条形码）上的寡核苷酸充当扩增替代物。然后使用 Scanomettic 方法检测条形码。该测定的检测限确定为 2.5 fM，这是用于检测双链基因组 DNA 的条形码型测定的首次演示。
The detection of bacterial genomic DNA through a nonenzymatic nanomaterials-based amplification method, the bio bar code assay, is reported. The assay utilizes oligonucleotide-functionalized magnetic microparticles to capture the target of interest from the sample. A critical step in the new assay involves the use of blocking oligonucleotides during heat denaturation of the double-stranded DNA. These blockers bind to specific regions of the target DNA upon cooling and prevent the duplex DNA from rehybridizing, which allows the particle probes to bind. Following target isolation using the magnetic particles, oligonucleotide-functionalized gold nanoparticles act as target recognition agents. The oligonucleotides on the nanoparticle (bar codes) act as amplification surrogates. The bar codes are then detected using the Scanomettic method. The limit of detection for this assay was determined to be 2.5 fM, and this is the first demonstration of a bar code-type assay for the detection of double-stranded, genomic DNA.