Department of Pharmacology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran; Student Research Committee, Mashhad University of Medical Sciences, Mashhad, Iran.
Medical Toxicology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Medical Biotechnology and Nanotechnology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Biosens Bioelectron. 2022 Jul 1;207:114209. doi: 10.1016/j.bios.2022.114209. Epub 2022 Mar 22.
The sudden increase of the COVID-19 outbreak and its continued growth with mutations in various forms has created a global health crisis as well as devastating social and economic effects over the past two years. In this study, a screen-printed carbon electrode reinforced with boron nitride quantum dots/flower-like gold nanostructures (BNQDs/FGNs/SPCE) and functionalized by highly specific antisense DNA oligonucleotide presents an alternative and promising solution for targeting SARS-CoV-2 RNA without nucleic acid amplification. The platform was tested on 120 SARS-CoV-2 RNA isolated from real clinical samples (60 positive and 60 negative confirmed by conventional RT-PCR method). Based on obtained quantitative results and statistical analysis (box-diagram, cutoff value, receiver operating characteristic curve, and t-test), the biosensor revealed a significant difference between the two positive and negative groups with 100% sensitivity and 100% specificity. To evaluate the quantitation capacity and detection limit of the biosensor for clinical trials, the detection performance of the biosensor for continuously diluted RNA isolated from SARS-CoV-2-confirmed patients was compared to those obtained by RT-PCR, demonstrating that the detection limit of the biosensor is lower than or comparable to that of RT-PCR. The ssDNA/BNQDs/FGNs/SPCE showed negligible cross-reactivity with RNA fragments isolated from Influenza A (IAV) clinical samples and also remained stable for up to 14 days. In conclusion, the fabricated biosensor may serve as a promising tool for point-of-care applications.
在过去的两年中，COVID-19 疫情的突然爆发及其以各种形式不断增长，造成了全球卫生危机以及毁灭性的社会和经济影响。在这项研究中，一种基于硼氮量子点/花状金纳米结构（BNQDs/FGNs/SPCE）增强的丝网印刷碳电极，并通过高度特异性的反义 DNA 寡核苷酸功能化，为靶向 SARS-CoV-2 RNA 提供了一种替代且有前途的解决方案，而无需进行核酸扩增。该平台在 120 个从真实临床样本中分离的 SARS-CoV-2 RNA 上进行了测试（60 个阳性，60 个通过常规 RT-PCR 方法证实的阴性）。基于获得的定量结果和统计分析（箱线图、截止值、受试者工作特征曲线和 t 检验），该生物传感器在两组阳性和阴性之间显示出 100%的灵敏度和 100%的特异性，具有显著差异。为了评估生物传感器在临床试验中的定量能力和检测限，将生物传感器对从 SARS-CoV-2 确诊患者中分离的连续稀释 RNA 的检测性能与 RT-PCR 进行了比较，结果表明，生物传感器的检测限低于或可与 RT-PCR 相媲美。ssDNA/BNQDs/FGNs/SPCE 与从流感 A（IAV）临床样本中分离的 RNA 片段几乎没有交叉反应性，并且在长达 14 天的时间内保持稳定。总之，该生物传感器可作为即时护理应用的有前途的工具。
