Methylene Blue functionalized carbon nanodots combined with different shape gold nanostructures for sensitive and selective SARS-CoV-2 sensing

The development of DNA-sensing platforms based on new synthetized Methylene Blue functionalized carbon nanodots combined with different shape gold nanostructures (AuNs), as a new pathway to develop a selective and sensitive methodology for SARS-CoV-2 detection is presented. A mixture of gold nanoparticles and gold nanotriangles have been synthetized to modify disposable electrodes that act as an enhanced nanostructured electrochemical surface for DNA probe immobilization. On the other hand, modified carbon nanodots prepared a la carte to contain Methylene Blue (MB-CDs) are used as electrochemical indicators of the hybridization event. These MB-CDs, due to their structure, are able to interact differently with double and single-stranded DNA molecules. Based on this strategy, target sequences of the SARS-CoV-2 virus have been detected in a straightforward way and rapidly with a detection limit of 2.00 aM. Moreover, this platform allows the detection of the SARS-CoV-2 sequence in the presence of other viruses, and also a single nucleotide polymorphism (SNPs). The developed approach has been tested directly on RNA obtained from nasopharyngeal samples from COVID-19 patients, avoiding any amplification process. The results agree well with those obtained by RT-qPCR or reverse transcription quantitative polymerase chain reaction technique.We acknowledge the support from the Comunidad de Madrid (TRANSNANOAVANSENS-CM, S2018/NMT-4349, NANOCOV-CM, SI3/PJI/2021–00341) and Ministerio de economia y competitividad de España (PID2020–116728RB-100, CTQ2015–71955-REDT (ELECTROBIONET)). IMDEA Nanociencia acknowledges support from the Programme for Centres of Excellence in R&D ‘Severo Ochoa’ (CEX2020–001039-S, MINECO). Authors also acknowledge REACT EU NANOCOV-CM project. RdC acknowledges support from Fundación IMDEA Nanociencia, Banco Santander, UAM (convocatoria CRUE- SANTANDER-CSIC, reference 10.01.03.02.41).Peer reviewe