Del Bonis-O'Donnell Jackson Travis, Fygenson Deborah K, Pennathur Sumita
Department of Mechanical Engineering, University of California Santa Barbara, Santa Barbara, CA, USA.
Analyst. 2015 Mar 7;140(5):1609-15. doi: 10.1039/c4an01735h.
DNA-stabilized fluorescent silver nanoclusters (AgNC DNA) are a new class of fluorophore that are formed by sequence specific interactions between silver and single-stranded DNA. By incorporating both target-binding and fluorescent-reporting sequences into a single synthetic DNA oligomer, AgNC DNA probes eliminate the need to conjugate dye or quencher molecules. In this study, we modify a AgNC DNA probe to demonstrate single-color multiplexed detection of DNA targets. We show that appending different lengths of poly-dT to the probe sequences tunes the electrophoretic mobility of AgNC DNA probes without affecting their fluorescence spectra. We use this to introduce a set of AgNC DNA probes selective for Hepatitis A, B and C target sequences that can be processed together in a simple, single-step protocol and distinguished with a resolution of 3.47 and signal to noise ratio of 17.23 in under 10 seconds by microfluidic capillary electrophoresis.
DNA稳定的荧光银纳米簇（AgNC DNA）是一类新型荧光团，由银与单链DNA之间的序列特异性相互作用形成。通过将靶标结合序列和荧光报告序列整合到单个合成DNA寡聚物中，AgNC DNA探针无需偶联染料或淬灭分子。在本研究中，我们对AgNC DNA探针进行了修饰，以展示对DNA靶标的单色多重检测。我们发现，在探针序列上附加不同长度的聚dT可调节AgNC DNA探针的电泳迁移率，而不影响其荧光光谱。我们利用这一点引入了一组对甲型、乙型和丙型肝炎靶标序列具有选择性的AgNC DNA探针，这些探针可以在一个简单的单步方案中一起处理，并通过微流控毛细管电泳在不到10秒的时间内以3.47的分辨率和17.23的信噪比进行区分。
