An integrated disposable device for DNA extraction and helicase dependent amplification.

An integrated disposable device for DNA extraction and helicase dependent amplification.
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DOI:
10.1007/s10544-009-9391-8
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发表时间:
2010-04
影响因子:
2.8
通讯作者:
Klapperich CM
Klapperich CM
中科院分区:
工程技术3区
文献类型:
--
作者:
Mahalanabis M;Do J;ALMuayad H;Zhang JY;Klapperich CM

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在这里,我们报告了一个集成的微流控芯片,进行解旋酶依赖性扩增(HDA)对含有活细菌的样品的演示。组合的基于芯片的样品制备和等温扩增对于世界卫生应用是有吸引力的,因为减少或消除了对控制流速和温度变化的仪器的需要。将细菌裂解、核酸提取和具有荧光报告物的DNA扩增并入一次性聚合物盒形式中。智能被动流体控制使用瓣阀和疏水排气口(具有纳米多孔PTFE膜)以及简单的片上混合器,消除了多个用户操作。该设备能够检测到少至10个菌落形成单位(CFU)的大肠杆菌。大肠杆菌中。
Here we report the demonstration of an integrated microfluidic chip that performs helicase dependent amplification (HDA) on samples containing live bacteria. Combined chip-based sample preparation and isothermal amplification are attractive for world health applications, since the need for instrumentation to control flow rate and temperature changes are reduced or eliminated. Bacteria lysis, nucleic acid extraction, and DNA amplification with a fluorescent reporter are incorporated into a disposable polymer cartridge format. Smart passive fluidic control using a flap valve and a hydrophobic vent (with a nanoporous PTFE membrane) with a simple on-chip mixer eliminates multiple user operations. The device is able to detect as few as ten colony forming units (CFU) of E. coli in growth medium.
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