Rüppel Nadine, Tröger Vicky, Sandetskaya Natalia, Kuhlmeier Dirk, Schmieder Stefan, Sonntag Frank
Fraunhofer Institute for Cell Therapy and Immunology IZI Leipzig Germany.
Institute of Analytical Chemistry University of Leipzig Leipzig Germany.
Eng Life Sci. 2018 Jan 29;18(4):263-268. doi: 10.1002/elsc.201700098. eCollection 2018 Apr.
In this work, an approach for SPR spectroscopy using the SPR system is examined that combines signal amplification by PCR and magnetic nanoparticles in one injection step. Therefore, the synthesis of PCR products was performed on the beads similar to a solid-phase PCR, termed PCR-on-a-bead. The functionality of this PCR was proven using an enzymatic assay. For validation the detection of oligonucleotides by SPR, an asymmetric PCR product was investigated. A signal increase upon binding of the PCR product to the specific probes was observed. In addition, surface regeneration of the chip was examined and reuse for at least two times ascertained. Amplification of the SPR signal by magnetic beads was verified but no signal was detected for PCR products immobilized on particles prior to injection.
在这项工作中，研究了一种使用表面等离子体共振（SPR）系统的SPR光谱方法，该方法在一个注射步骤中结合了通过聚合酶链反应（PCR）和磁性纳米颗粒进行的信号放大。因此，PCR产物的合成在类似于固相PCR的珠子上进行，称为珠上PCR。使用酶促测定法证明了这种PCR的功能。为了验证通过SPR对寡核苷酸的检测，研究了一种不对称PCR产物。观察到PCR产物与特异性探针结合后信号增加。此外，还检查了芯片的表面再生情况，并确定至少可重复使用两次。验证了磁性珠对SPR信号的放大作用，但在注射前未检测到固定在颗粒上的PCR产物的信号。
