Drori Paz, Mouhadeb Odelia, Moya Muñoz Gabriel G, Razvag Yair, Alcalay Ron, Klocke Philipp, Cordes Thorben, Zahavy Eran, Lerner Eitan
Department of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, Faculty of Mathematics & Science, The Edmond J. Safra Campus, The Hebrew University of Jerusalem, Jerusalem 9190401, Israel.
Department of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness Ziona, Israel.
iScience. 2024 Sep 17;27(10):110982. doi: 10.1016/j.isci.2024.110982. eCollection 2024 Oct 18.
Mainstream virus detection relies on the specific amplification of nucleic acids via polymerase chain reaction, a process that is slow and requires extensive laboratory expertise and equipment. Other modalities, such as antigen-based tests, allow much faster virus detection but have reduced sensitivity. In this study, we introduce an approach for rapid and specific detection of single nanoparticles using a confocal-based flow virometer. The combination of laminar flow in a microfluidic channel and correlated fluorescence signals emerging from both free dyes and fluorescently labeled primary antibodies provide insights into nanoparticle volumes and specificities. We evaluate and validate the assay using fluorescent beads and viruses, including SARS-CoV-2 with fluorescently labeled primary antibodies. Additionally, we demonstrate how hydrodynamic focusing enhances the assay sensitivity for detecting viruses at relevant loads. Based on our results, we envision the future use of this technology for clinically relevant bio-nanoparticles, supported by the implementation of the assay in a portable and user-friendly setup.
主流病毒检测依赖于通过聚合酶链反应对核酸进行特异性扩增，这一过程缓慢，且需要广泛的实验室专业知识和设备。其他检测方式，如基于抗原的检测，能实现更快的病毒检测，但灵敏度较低。在本研究中，我们介绍了一种使用基于共聚焦的流动病毒仪快速、特异性检测单个纳米颗粒的方法。微流控通道中的层流与游离染料和荧光标记的一抗产生的相关荧光信号相结合，可深入了解纳米颗粒的体积和特异性。我们使用荧光微球和病毒（包括用荧光标记一抗标记的严重急性呼吸综合征冠状病毒2）对该检测方法进行评估和验证。此外，我们还展示了流体动力聚焦如何提高在相关载量下检测病毒的检测灵敏度。基于我们的研究结果，我们设想在便携式且用户友好的装置中实施该检测方法，以支持该技术未来用于临床相关生物纳米颗粒。
