Trypsin-amplified Aerolysin Nanopore Amplified Sandwich Assay for Attomolar Nucleic Acid and Single Bacteria Detection

Nanopore technology is promising for the next-generation of nucleic acid-based diagnosis. However, sequence resolvation could still be hardly achieved in low-concentration. Herein, we propose a tryp-sin-activated catalysis reaction for amplified detection, which substantially improves the sensitiv-ity of nanopore technique. The proposed trypsin-amplified nanopore amplified sandwich assay (tNASA) could contribute to a sensitivity approximately 1,00,000 times higher based on nucleic acid probe design. Remarkably, tNASA is capable of attomolar nucleic acid and single cell detection by using a miniaturized current amplifier without alignment algorithm. And it allows to accurately and robustly profiling 10 pathogenic species in serum, thus utilized for the diagnosis of infectious diseases. tNASA may evolve the construction of nanopore techniques for nucleic acid detection, would facili-tate its translation for pocket diagnosis and precision medicine.