On-chip conductometric detection of short DNA sequences via electro-hydrodynamic aggregation†,Analyst

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On-chip conductometric detection of short DNA sequences via electro-hydrodynamic aggregation†
Analyst ( IF 3.6 ) Pub Date : 2017-11-24 00:00:00 , DOI: 10.1039/c7an00798a
B. Venzac _{1,

2,

3,

4,

5} , M. L. Diakité _{1,

2,

3,

4,

5} , D. Herthnek _{6,

7,

8,

9,

10} , I. Cissé _{11,

12,

13,

14,

15} , U. Bockelmann _{11,

12,

13,

14,

15} , S. Descroix _{1,

2,

3,

4,

5} , L. Malaquin _{1,

2,

3,

4,

5} , J.-L. Viovy _{1,

2,

3,

4,

5}

Affiliation

Fluorescence measurement is the main technology for post-amplification DNA detection in automated systems. Direct electrical reading of DNA concentration in solution could be an interesting alternative to go toward more miniaturized or less expensive devices, in particular in the pathogen detection field. Here we present the detection of short bacterial biomarkers with a direct impedancemetric measurement, within solutions of amplified and elongated DNA sequences in a microchannel. This technology relies on the electrohydrodynamic instability occurring in solutions of long charged macromolecules in a strong electric field. This instability specifically induces the aggregation of long DNAs and triggers conductivity variations that can be monitored by on-contact conductometry. An innovative isothermal amplification and elongation strategy was developed, combining SDA and HRCA reactions, in order to yield long DNAs suitable to be detected by the above principle, from a dilute initial DNA target. In contrast with previous label-free detection methods, this new strategy is very robust to matrix effects, thanks to the unique molecular weight dependence of the instability, coupled with this specific DNA amplification strategy. We demonstrate the detection of a 1 pM gene sequence specific to Staphylococcus aureus, in a portable system.

中文翻译：

通过电-液动力聚合对短DNA序列进行片上电导检测†

荧光测量是自动化系统中扩增后DNA检测的主要技术。直接电读取溶液中DNA的浓度可能是走向更小型或更便宜的设备的一个有趣的选择，特别是在病原体检测领域。在这里，我们介绍了在微通道中扩增和延长的DNA序列溶液中，用直接阻抗测量法检测短细菌生物标志物的方法。该技术依赖于在强电场中长电荷大分子溶液中发生的电流体动力学不稳定性。这种不稳定性会特异性地引起长DNA的聚集，并触发电导率变化，可以通过接触电导法进行监测。开发了一种创新的等温扩增和延伸策略，结合了SDA和HRCA反应，以从稀释的初始DNA靶标产生适合通过上述原理检测的长DNA。与以前的无标记检测方法相比，由于不稳定性具有独特的分子量依赖性，再加上这种特定的DNA扩增策略，因此这种新策略对基质效应非常鲁棒。我们证明了对1 pM基因序列的检测加上这种特定的DNA扩增策略。我们证明了对1 pM基因序列的检测加上这种特定的DNA扩增策略。我们证明了对1 pM基因序列的检测金黄色葡萄球菌，在便携式系统中。

更新日期：2017-11-24

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