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Low-cost inkjet-printed nanostructured biosensor based on CRISPR/Cas12a system for pathogen detection
Biosensors and Bioelectronics ( IF 12.6 ) Pub Date : 2024-04-25 , DOI: 10.1016/j.bios.2024.116340 Angela Gilda Carota , Andrea Bonini , Massimo Urban , Noemi Poma , Federico Maria Vivaldi , Arianna Tavanti , Marianna Rossetti , Giulio Rosati , Arben Merkoçi , Fabio Di Francesco
Biosensors and Bioelectronics ( IF 12.6 ) Pub Date : 2024-04-25 , DOI: 10.1016/j.bios.2024.116340 Angela Gilda Carota , Andrea Bonini , Massimo Urban , Noemi Poma , Federico Maria Vivaldi , Arianna Tavanti , Marianna Rossetti , Giulio Rosati , Arben Merkoçi , Fabio Di Francesco
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The escalating global incidence of infectious diseases caused by pathogenic bacteria, especially in developing countries, emphasises the urgent need for rapid and portable pathogen detection devices. This study introduces a sensitive and specific electrochemical biosensing platform utilising cost-effective electrodes fabricated by inkjet-printing gold and silver nanoparticles on a plastic substrate. The biosensor exploits the CRISPR/Cas12a system for detecting a specific DNA sequence selected from the genome of the target pathogen. Upon detection, the trans-activity of Cas12a/gRNA is triggered, leading to the cleavage of rationally designed single-strand DNA reporters (linear and hairpin) labelled with methylene blue (ssDNA-MB) and bound to the electrode surface. In principle, this sensing mechanism can be adapted to any bacterium by choosing a proper guide RNA to target a specific sequence of its DNA. The biosensor's performance was assessed for two representative pathogens (a Gram-negative, , and a Gram-positive, ), and results obtained with inkjet-printed gold electrodes were compared with those obtained by commercial screen-printed gold electrodes. Our results show that the use of inkjet-printed nanostructured gold electrodes, which provide a large surface area, in combination with the use of hairpin reporters containing a poly-T loop can increase the sensitivity of the assay corresponding to a signal variation of 86%. DNA targets amplified from various clinically isolated bacteria, have been tested and demonstrate the potential of the proposed platform for point-of-need applications.
更新日期：2024-04-25
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