DESTINA Genomica S.L. Parque Tecnológico Ciencias de la Salud (PTS), Avenida de la Innovación 1, Edificio BIC, 18016, Armilla, Granada, Spain; GENYO Centre for Genomics and Oncological Research, Pfizer/University of Granada/Andalusian Regional Government, PTS Granada - Avenida de la Ilustración, 114- 18016, Granada, Spain; Department of Medicinal & Organic Chemistry, Faculty of Pharmacy, University of Granada, Campus de Cartuja s/n, Granada, Spain; Unit of Excellence in Chemistry Applied to Biomedicine and the Environment of the University of Granada, Granada, Spain; Instituto de Investigación Biosanitaria ibs.GRANADA, Granada, Spain.
DESTINA Genomica S.L. Parque Tecnológico Ciencias de la Salud (PTS), Avenida de la Innovación 1, Edificio BIC, 18016, Armilla, Granada, Spain.
Biosens Bioelectron. 2023 Jun 15;230:115268. doi: 10.1016/j.bios.2023.115268. Epub 2023 Mar 30.
The COVID-19 pandemic has highlighted the need for innovative approaches to its diagnosis. Here we present CoVradar, a novel and simple colorimetric method that combines nucleic acid analysis with dynamic chemical labeling (DCL) technology and the Spin-Tube device to detect SARS-CoV-2 RNA in saliva samples. The assay includes a fragmentation step to increase the number of RNA templates for analysis, using abasic peptide nucleic acid probes (DGL probes) immobilized to nylon membranes in a specific dot pattern to capture RNA fragments. Duplexes are formed by labeling complementary RNA fragments with biotinylated SMART bases, which act as templates for DCL. Signals are generated by recognizing biotin with streptavidin alkaline phosphatase and incubating with a chromogenic substrate to produce a blue precipitate. CoVradar results are analysed by CoVreader, a smartphone-based image processing system that can display and interpret the blotch pattern. CoVradar and CoVreader provide a unique molecular assay capable of detecting SARS-CoV-2 viral RNA without the need for extraction, preamplification, or pre-labeling steps, offering advantages in terms of time (∼3 h/test), cost (∼€1/test manufacturing cost) and simplicity (does not require large equipment). This solution is also promising for developing assays for other infectious diseases.
COVID-19 大流行凸显了创新方法诊断的必要性。在这里,我们提出了 CoVradar,这是一种新颖而简单的比色法,将核酸分析与动态化学标记(DCL)技术和 Spin-Tube 装置相结合,用于检测唾液样本中的 SARS-CoV-2 RNA。该测定包括一个片段化步骤,以增加用于分析的 RNA 模板数量,使用固定在尼龙膜上的碱性肽核酸探针(DGL 探针)以特定的点图案捕获 RNA 片段。通过用生物素化 SMART 碱基标记互补的 RNA 片段形成双链体,这些碱基充当 DCL 的模板。通过用链霉亲和素碱性磷酸酶识别生物素并与显色底物孵育来产生蓝色沉淀来产生信号。CoVradar 结果由基于智能手机的图像处理系统 CoVreader 进行分析,该系统可以显示和解释斑点图案。CoVradar 和 CoVreader 提供了一种独特的分子测定法,能够检测 SARS-CoV-2 病毒 RNA,而无需提取、预扩增或预标记步骤,在时间(约 3 小时/测试)、成本(约 1 欧元/测试制造成本)和简单性(不需要大型设备)方面具有优势。该解决方案对于开发其他传染病的测定法也很有前途。