Liu Changchun, Mauk Michael G, Bau Haim H
Department of Mechanical Engineering and Applied Mechanics, University of Pennsylvania, 229 Towne Building, 220 S. 33rd St., Philadelphia, PA 19104-6315 USA.
Microfluid Nanofluidics. 2011;11(2):209-220. doi: 10.1007/s10404-011-0788-3. Epub 2011 Mar 18.
An inexpensive, disposable, integrated, polymer-based cassette for loop-mediated isothermal amplification (LAMP) of target nucleic acids was designed, fabricated, and tested. The LAMP chamber was equipped with single-use, thermally actuated valves made with a composite consisting of a mixture of PDMS and expandable microspheres. The effect of the composite composition on its expansion was investigated, and the valve's performance was evaluated. In its closed state, the valve can hold pressures as high as 200 kPa without any significant leakage. Both the LAMP chamber and the valves were actuated with thin film heaters. The utility of the cassette was demonstrated by carrying out LAMP of DNA target and reverse transcribed loop meditated isothermal amplification (RT-LAMP) of RNA targets. The amplicons were detected in real time with a portable, compact detector. The system was capable of detecting as few as 10 target molecules per sample in well under 1 h. The portable, integrated cassette system described here is particularly suited for applications at the point of care and in resource-poor countries, where funds and trained personnel are in short supply.
设计、制造并测试了一种用于靶核酸环介导等温扩增（LAMP）的廉价、一次性、集成的聚合物基盒式装置。LAMP腔室配备有由聚二甲基硅氧烷（PDMS）和可膨胀微球混合物组成的复合材料制成的一次性热驱动阀。研究了复合材料组成对其膨胀的影响，并评估了阀门的性能。在关闭状态下，该阀能够承受高达200 kPa的压力而无明显泄漏。LAMP腔室和阀门均由薄膜加热器驱动。通过对DNA靶标进行LAMP以及对RNA靶标进行逆转录环介导等温扩增（RT-LAMP），证明了该盒式装置的实用性。使用便携式紧凑型检测器实时检测扩增子。该系统能够在不到1小时的时间内检测出每个样品中低至10个靶标分子。这里描述的便携式集成盒式系统特别适用于即时护理应用以及资源匮乏国家，因为这些地方资金和受过培训的人员短缺。
