Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, Università degli Studi di Milano, Segrate, Italy.
Proxentia S.r.l., viale Ortles 22/4, Milano, Italy.
Biosens Bioelectron. 2021 Jan 15;172:112751. doi: 10.1016/j.bios.2020.112751. Epub 2020 Oct 22.
MicroRNAs are widely studied as circulating biomarkers for early stage diagnosis of several diseases. Detection and quantification of miRNAs is currently performed through complex and time consuming procedures. Herein we demonstrate a rapid, multiplex, one-pot detection method based on two-step amplification of the signal measured by Reflective Phantom Interface (RPI) label-free optical biosensor. We achieved sub-pM quantification of different miRNAs in about 1.5 h, through specific capture with surface DNA probes combined to a 35-fold mass amplification by an antibody targeting DNA-RNA hybrids and polyclonal secondary antibody, all performed without washing steps. The assay is the result of a modelling and optimization of the multi-step process that has been made possible by the RPI characterization of each individual interaction involved.
微 RNA 被广泛研究作为几种疾病早期诊断的循环生物标志物。miRNA 的检测和定量目前通过复杂且耗时的程序来完成。在此，我们展示了一种基于两步放大的快速、多重、一体化检测方法，该方法通过反射幻影界面 (RPI) 无标记光学生物传感器测量的信号进行检测和定量。我们通过表面 DNA 探针的特异性捕获，并结合针对 DNA-RNA 杂交体的抗体和多克隆二级抗体进行 35 倍质量放大，在大约 1.5 小时内实现了不同 miRNA 的亚皮摩尔级定量，所有步骤均无需洗涤。该检测方法是对多步过程进行建模和优化的结果，这得益于 RPI 对每个涉及的相互作用的单独特征描述。
