Department of Energy and Refrigerating Air-Conditioning Engineering, National Taipei University of Technology, Taipei 10608, Taiwan.
Sensors (Basel). 2010;10(1):146-66. doi: 10.3390/s100100146. Epub 2009 Dec 28.
The chip-based polymerase chain reaction (PCR) system has been developed in recent years to achieve DNA quantification. Using a microstructure and miniature chip, the volume consumption for a PCR can be reduced to a nano-liter. With high speed cycling and a low reaction volume, the time consumption of one PCR cycle performed on a chip can be reduced. However, most of the presented prototypes employ commercial fluorimeters which are not optimized for fluorescence detection of such a small quantity sample. This limits the performance of DNA quantification, especially low experiment reproducibility. This study discusses the concept of a chip-oriented fluorimeter design. Using the analytical model, the current study analyzes the sensitivity and dynamic range of the fluorimeter to fit the requirements for detecting fluorescence in nano-liter volumes. Through the optimized processes, a real-time PCR on a chip system with only one nano-liter volume test sample is as sensitive as the commercial real-time PCR machine using the sample with twenty micro-liter volumes. The signal to noise (S/N) ratio of a chip system for DNA quantification with hepatitis B virus (HBV) plasmid samples is 3 dB higher. DNA quantification by the miniature chip shows higher reproducibility compared to the commercial machine with respect to samples of initial concentrations from 10(3) to 10(5) copies per reaction.
近年来，基于芯片的聚合酶链反应（PCR）系统已被开发出来以实现 DNA 定量。利用微结构和微型芯片，可以将 PCR 的体积消耗减少到纳升。通过高速循环和低反应体积，可以减少在芯片上进行的一个 PCR 循环的时间消耗。然而，大多数提出的原型采用的是商业荧光计，这些荧光计没有针对如此小体积的样品的荧光检测进行优化。这限制了 DNA 定量的性能，尤其是实验的低重复性。本研究讨论了面向芯片的荧光计设计的概念。利用分析模型，本研究分析了荧光计的灵敏度和动态范围，以满足检测纳升体积荧光的要求。通过优化工艺，仅使用一个纳升体积测试样品的实时芯片上 PCR 系统与使用二十微升体积样品的商用实时 PCR 机器一样灵敏。用于乙型肝炎病毒（HBV）质粒样品的 DNA 定量的芯片系统的信噪比（S/N）比高 3dB。与商用机器相比，微型芯片的 DNA 定量对于初始浓度为 10(3)到 10(5)个拷贝/反应的样品具有更高的重现性。
