Department of Chemistry, University of Virginia, McCormick Road, P.O. Box 400319, Charlottesville, VA 22904, USA.
Analyst. 2011 May 7;136(9):1928-37. doi: 10.1039/c0an00922a. Epub 2011 Mar 21.
A valveless microdevice has been developed for the integration of solid phase extraction (SPE) and polymerase chain reaction (PCR) on a single chip for the short tandem repeat (STR) analysis of DNA from a biological sample. The device consists of two domains--a SPE domain filled with silica beads as a solid phase and a PCR domain with an ~500 nL reaction chamber. DNA from buccal swabs was purified and amplified using the integrated device and a full STR profile (16 loci) resulted. The 16 loci Identifiler® multiplex amplification was performed using a non-contact infrared (IR)-mediated PCR system built in-house, after syringe-driven SPE, providing an ~80-fold and 2.2-fold reduction in sample and reagent volumes consumed, respectively, as well as an ~5-fold reduction in the overall analysis time in comparison to conventional analysis. Results indicate that the SPE-PCR system can be used for many applications requiring genetic analysis, and the future addition of microchip electrophoresis (ME) to the system would allow for the complete processing of biological samples for forensic STR analysis on a single microdevice.
已经开发出一种无阀微器件,用于将固相萃取 (SPE) 和聚合酶链反应 (PCR) 集成在单个芯片上,用于对生物样品中的 DNA 进行短串联重复序列 (STR) 分析。该器件由两个区域组成 - SPE 区域填充有作为固相的硅胶珠,以及具有约 500nL 反应室的 PCR 区域。使用集成器件对口腔拭子中的 DNA 进行纯化和扩增,得到完整的 STR 图谱 (16 个基因座)。使用内部构建的非接触式红外 (IR) 介导 PCR 系统进行 16 个基因座 Identifiler®多重扩增,在注射器驱动 SPE 之后,分别使样品和试剂体积消耗减少约 80 倍和 2.2 倍,以及总分析时间减少约 5 倍与传统分析相比。结果表明,SPE-PCR 系统可用于许多需要遗传分析的应用,并且将来向系统中添加微芯片电泳 (ME) 将允许在单个微器件上完成对法医 STR 分析的生物样品的完整处理。