Sequence-specific DNA detection at 10 fM by electromechanical signal transduction.
Sequence-specific DNA detection at 10 fM by electromechanical signal transduction.
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DOI：
10.1021/ac5021408
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发表时间：
2014-10-07
影响因子：
7.4
通讯作者：
Schmidt, Jacob J.
中科院分区：
文献类型：
--
作者：
Esfandiari, Leyla;Lorenzini, Michael;Kocharyan, Gayane;Monbouquette, Harold G.;Schmidt, Jacob J.
在模拟真实样本中存在的非互补基因组DNA的DNA背景下，使用一种基于机电信号转导的简单、无PCR、无光学的方法检测到10fM浓度(约6×105个分子)的目标DNA片段。开发一种快速、灵敏和经济高效的核酸检测平台是人们迫切需要的，以满足一系列不同的应用。我们之前描述了一种潜在的低成本的序列特异性核酸检测设备，该设备基于与目标核酸杂交时被电泳化的微球(多肽核酸探针)偶联物堵塞的孔的电导变化测量。在这里，我们演示了我们的设备使用更长的DNA靶标的操作，并描述了由此产生的检测极限(LOD)的改进。我们研究了在下午1点到1fm检测110,235,419和1613个核苷酸的寡聚体，发现LOD随着DNA长度的增加而减少，419和1613个核苷酸寡聚体可以检测到10fm。此外，对于长度相似的非互补对照DNA片段，没有获得假阳性反应。1613碱基的DNA寡聚体在大小上与16S rRNA相似，这表明我们的设备可能有助于基于对物种特异性16S rRNA序列的识别来检测临床相关浓度的病原菌。
Target DNA fragments at 10 fM concentration (approximately 6 × 105 molecules) were detected against a DNA background simulating the noncomplementary genomic DNA present in real samples using a simple, PCR-free, optics-free approach based on electromechanical signal transduction. The development of a rapid, sensitive, and cost-effective nucleic acid detection platform is highly desired for a range of diverse applications. We previously described a potentially low-cost device for sequence-specific nucleic acid detection based on conductance change measurement of a pore blocked by electrophoretically mobilized bead-(peptide nucleic acid probe) conjugates upon hybridization with target nucleic acid. Here, we demonstrate the operation of our device with longer DNA targets, and we describe the resulting improvement in the limit of detection (LOD). We investigated the detection of DNA oligomers of 110, 235, 419, and 1613 nucleotides at 1 pM to 1 fM and found that the LOD decreased as DNA length increased, with 419 and 1613 nucleotide oligomers detectable down to 10 fM. In addition, no false positive responses were obtained with noncomplementary, control DNA fragments of similar length. The 1613-base DNA oligomer is similar in size to 16S rRNA, which suggests that our device may be useful for detection of pathogenic bacteria at clinically relevant concentrations based on recognition of species-specific 16S rRNA sequences.
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影响因子：
5.1
作者：
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通讯作者：
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影响因子：
4.3
作者：
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通讯作者：
Niessner R
影响因子：
15
作者：
Esfandiari, Leyla;Monbouquette, Harold G.;Schmidt, Jacob J.
通讯作者：
Schmidt, Jacob J.
影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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