A Single Electrochemical Probe Used for Analysis of Multiple Nucleic Acid Sequences.
A Single Electrochemical Probe Used for Analysis of Multiple Nucleic Acid Sequences.
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用于分析多核酸序列的单一电化学探针。
DOI：
10.1002/elan.201600548
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发表时间：
2017-03
期刊：
影响因子：
3
通讯作者：
Chumbimuni-Torres KY
中科院分区：
文献类型：
--
作者：
Mills DM;Calvo-Marzal P;Pinzon JM;Armas S;Kolpashchikov DM;Chumbimuni-Torres KY
电化学杂交传感器已被广泛地探索用于分析特定的核酸。然而，该平台的商业化受到需要将与RNA或DNA靶互补的单独的寡核苷酸探针附着到电极表面的阻碍。在这里，我们证明了一个单一的探针可以用于分析几个核酸目标，具有高选择性和低成本。通用电化学四向结（4J）形成（UE 4J）传感器由连接到电极表面的通用DNA茎环（USL）探针和两个适配器链（m和f）组成，所述适配器链与USL探针和分析物杂交以形成4J缔合物。将m衔接子链与亚甲蓝氧化还原标记物缀合用于信号ON感测，并使用方波伏安法监测。我们证明了单个传感器可以用于检测几种不同的DNA/RNA序列，并且可以通过简单的水冲洗在30秒内再生。即使在室温下，UE 4J传感器也能通过识别单个碱基取代实现高选择性。UE 4J传感器为可重复使用的通用平台开辟了一个场所，该平台可以低成本用于分析DNA或RNA目标。
Electrochemical hybridization sensors have been explored extensively for analysis of specific nucleic acids. However, commercialization of the platform is hindered by the need for attachment of separate oligonucleotide probes complementary to a RNA or DNA target to an electrode’s surface. Here we demonstrate that a single probe can be used to analyze several nucleic acid targets with high selectivity and low cost. The universal electrochemical four-way junction (4J)-forming (UE4J) sensor consists of a universal DNA stem-loop (USL) probe attached to the electrode’s surface and two adaptor strands (m and f) which hybridize to the USL probe and the analyte to form a 4J associate. The m adaptor strand was conjugated with a methylene blue redox marker for signal ON sensing and monitored using square wave voltammetry. We demonstrated that a single sensor can be used for detection of several different DNA/RNA sequences and can be regenerated in 30 seconds by a simple water rinse. The UE4J sensor enables a high selectivity by recognition of a single base substitution, even at room temperature. The UE4J sensor opens a venue for a re-useable universal platform that can be adopted at low cost for the analysis of DNA or RNA targets.
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