Department of Biophysics, School of Medicine, Yeditepe University, Istanbul, Turkey.
Department of Medical Biology, School of Medicine, Yeditepe University, Istanbul, Turkey.
Talanta. 2022 Aug 15;246:123429. doi: 10.1016/j.talanta.2022.123429. Epub 2022 Apr 2.
Viral infection has been one of the major health issues for human life. The real-time reverse transcription polymerase chain reaction (RT-PCR)-based detection has primarily been used for virus detection as a highly reliable procedure. However, it is a relatively long and multi-stage process. In addition, required skilled personnel and complex instrumentation presents difficulties in large scale monitoring efforts. Therefore, we report here a direct and fast detection method for CoV-2 genome as applied in the nose-throat swab samples without any further processing. The detection principle is based on fluorescein-loaded mesoporous silica nanoparticles capped by specific gene sequences probes immobilized on the surface of the nanoparticles. Upon hybridization with the target viral genome, the fluorescein molecules were released from the mesopores. Testing with synthetic oligonucleotides, the NSP12 gene-based detection resulted in a strong signal. Target detection time could be optimized to 15 min and the limit of detection was 1.4 RFU with 84% sensitivity with clinical samples (n = 43).
病毒感染一直是人类生命的主要健康问题之一。基于实时逆转录聚合酶链反应 (RT-PCR) 的检测主要用于病毒检测，是一种高度可靠的方法。然而，它是一个相对较长且多阶段的过程。此外，所需的熟练人员和复杂的仪器在大规模监测工作中带来了困难。因此，我们在此报告了一种直接且快速的 CoV-2 基因组检测方法，应用于未经进一步处理的鼻喉拭子样本。检测原理基于负载荧光素的介孔硅纳米粒子，其表面固定有特定基因序列探针。与目标病毒基因组杂交后，荧光素分子从介孔中释放出来。用合成寡核苷酸进行测试，基于 NSP12 基因的检测产生了强烈的信号。目标检测时间可以优化至 15 分钟，检测限为 1.4 RFU，临床样本的灵敏度为 84%（n=43）。
