Rapid and simple single-chamber nucleic acid detection system prepared through nature-inspired surface engineering.
Rapid and simple single-chamber nucleic acid detection system prepared through nature-inspired surface engineering.
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作者：
Park J;Woo S;Kim J;Lee H;Yoo YE;Hong S
背景资料：基于核酸（NA）的诊断能够对各种疾病做出快速反应，但目前的技术通常需要多个劳动密集型步骤，这是成功转化为临床环境的主要障碍。方法：我们报道了一种表面工程单室装置，用于NA提取和原位扩增，无需样品转移。我们的系统有两个反应位点：一个NA提取室，其表面用微柱图案化，一个反应室填充有用于基于原位聚合酶的NA扩增的试剂。这两个网站集成在一个单一的微流体装置;我们应用塑料注塑成型的成本效益，大规模生产的设计设备。微柱通过自然灵感的二氧化硅涂层进行化学活化，以具有对NA的特异性亲和力。结果如下：作为概念验证，将比色pH指示剂与NA的芯片上分析结合，以快速方便地检测病原体。NA富集效率取决于裂解物孵育时间，因为扩散控制NA与工程化表面的接触。我们可以通过肉眼在总测定时间的1小时内检测到低至1 × 103 CFU。结论：我们预计，NA富集的表面工程技术可以很容易地集成为各种类型的微流控芯片的一部分，用于快速和方便的基于核酸的诊断。
Background: Nucleic acid (NA)-based diagnostics enable a rapid response to various diseases, but current techniques often require multiple labor-intensive steps, which is a major obstacle to successful translation to a clinical setting. Methods: We report on a surface-engineered single-chamber device for NA extraction and in situ amplification without sample transfer. Our system has two reaction sites: a NA extraction chamber whose surface is patterned with micropillars and a reaction chamber filled with reagents for in situ polymerase-based NA amplification. These two sites are integrated in a single microfluidic device; we applied plastic injection molding for cost-effective, mass-production of the designed device. The micropillars were chemically activated via a nature-inspired silica coating to possess a specific affinity to NA. Results: As a proof-of-concept, a colorimetric pH indicator was coupled to the on-chip analysis of NA for the rapid and convenient detection of pathogens. The NA enrichment efficiency was dependent on the lysate incubation time, as diffusion controls the NA contact with the engineered surface. We could detect down to 1×103 CFU by the naked eye within one hour of the total assay time. Conclusion: We anticipate that the surface engineering technique for NA enrichment could be easily integrated as a part of various types of microfluidic chips for rapid and convenient nucleic acid-based diagnostics.
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DOI：
10.1016/j.btre.2016.10.001
发表时间：
2016-12
期刊：
Biotechnology reports (Amsterdam, Netherlands)
影响因子：
--
作者：
El-Ashram S;Al Nasr I;Suo X
通讯作者：
Suo X
影响因子：
5.4
作者：
Hong, Seonki;Yeom, Jihyeon;Lee, Haeshin
通讯作者：
Lee, Haeshin
影响因子：
2.8
作者：
Barra, Gustavo Barcelos;Santa Rita, Ticiane Henriques;Soares Costa, Sandra Santana
通讯作者：
Soares Costa, Sandra Santana
影响因子：
1.8
作者：
Yaffe H;Buxdorf K;Shapira I;Ein-Gedi S;Moyal-Ben Zvi M;Fridman E;Moshelion M;Levy M
通讯作者：
Levy M
影响因子：
64.5
作者：
Snyder MW;Kircher M;Hill AJ;Daza RM;Shendure J
通讯作者：
Shendure J