Institute of Photonics and Electronics of the Czech Academy of Sciences, Chaberská 1014/57, 182 51 Prague, Czech Republic.
Institute of Hematology and Blood Transfusion, U Nemocnice 2094/1, 128 20 Prague, Czech Republic.
Biosens Bioelectron. 2021 Dec 15;194:113613. doi: 10.1016/j.bios.2021.113613. Epub 2021 Sep 3.
Microribonucleic acids (miRNAs) are short noncoding ribonucleic acids that have been linked with a multitude of human diseases including lung, breast, and hematological cancers. In this work, we present a novel, extremely sensitive assay for the label-free optical biosensor-based detection of miRNAs, which is based on the oligonucleotide-triggered release of nanoparticles from a sensor surface. We combine this assay (herein referred to as the nanoparticle-release (NPR) assay) with a surface plasmon resonance biosensor and show that the assay is able to enhance the specific sensor response associated with the binding of target miRNA while suppressing the interfering effects caused by the non-specific binding. We apply the assay to the detection of miRNAs related to myelodysplastic syndromes (miR-125b, miR-16) in blood plasma and demonstrate that the assay enables detection of miR-125b with a limit of detection (LOD) of 349 aM (corresponding to the lowest detectable amounts of 419 zmol). The achieved LOD is better by a factor of ∼100 when compared to the conventional nanoparticle-enhanced sandwich assay. Moreover, we demonstrate that the NPR assay may be combined with time-division multiplexing for the multiplexed miRNA detection.
微小核糖核酸（miRNAs）是短链非编码核糖核酸，与包括肺癌、乳腺癌和血液癌症在内的多种人类疾病有关。在这项工作中，我们提出了一种新颖的、极其灵敏的基于无标记光学生物传感器的 miRNA 检测方法，该方法基于寡核苷酸触发从传感器表面释放纳米粒子。我们将该测定法（在此称为纳米粒子释放（NPR）测定法）与表面等离子体共振生物传感器结合，并表明该测定法能够增强与靶 miRNA 结合相关的特定传感器响应，同时抑制由非特异性结合引起的干扰效应。我们将该测定法应用于检测与骨髓增生异常综合征相关的 miRNA（miR-125b、miR-16）在血浆中的表达情况，并证明该测定法能够以 349 aM 的检测限（LOD）检测 miR-125b（相当于 419 zmol 的最低可检测量）。与传统的纳米粒子增强型三明治测定法相比，该测定法的检测限提高了约 100 倍。此外，我们证明，NPR 测定法可以与分时复用结合用于多重 miRNA 检测。
