Hou Chih-Sheng Johnson, Milovic Nebojsa, Godin Michel, Russo Peter R, Chakrabarti Raj, Manalis Scott R
Department of Electrical Engineering and Computer Science, Biological Engineering Division, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Anal Chem. 2006 Apr 15;78(8):2526-31. doi: 10.1021/ac0520689.
We present a robust and simple method for direct, label-free PCR product quantification using an integrated microelectronic sensor. The field-effect sensor can sequentially detect the intrinsic charge of multiple unprocessed PCR products and does not require sample processing or additional reagents in the PCR mixture. The sensor measures nucleic acid concentration in the PCR relevant range and specifically detects the PCR products over reagents such as Taq polymerase and nucleotide monomers. The sensor can monitor the product concentration at various stages of PCR and can generate a readout that resembles that of a real-time fluorescent measurement using an intercalating dye but without its potential inhibition artifacts. The device is mass-produced using standard semiconductor processes, can be reused for months, and integrates all sensing components directly on-chip. As such, our approach establishes a foundation for the direct integration of PCR-based in vitro biotechnologies with microelectronics.
我们展示了一种使用集成微电子传感器进行直接、无标记PCR产物定量的强大且简单的方法。场效应传感器可以顺序检测多个未处理的PCR产物的固有电荷，并且不需要对样品进行处理或在PCR混合物中添加额外试剂。该传感器可在与PCR相关的范围内测量核酸浓度，并能特异性地检测PCR产物，而不受诸如Taq聚合酶和核苷酸单体等试剂的干扰。该传感器可以监测PCR各个阶段的产物浓度，并能生成类似于使用嵌入染料进行实时荧光测量的读数，但不会产生潜在的抑制假象。该设备采用标准半导体工艺进行大规模生产，可重复使用数月，并且直接在芯片上集成了所有传感组件。因此，我们的方法为基于PCR的体外生物技术与微电子的直接集成奠定了基础。
