Recently, the Precision Medicine Single-Molecule Diagnostics Technology Research Center at Chongqing Institute of Green and Intelligent Technology, Chinese Academy of Sciences has achieved a series of progress in bacterial endotoxin, oncogene conformation identification and protein-nucleic acid specific interaction monitoring based on solid-state pore devices. The results were published in: "Biosens. Bioelectron.", "Chem. Eng. J.", "ACS Appl. Mater. Interf. " and "Coor. Chem. Rev. ".

Bacterial endotoxin of Gram-negative bacteria is the main cause of urinary dysfunction and sepsis. The detection of endotoxin lipopolysaccharide reported in the literature is mainly based on the light, heat and electrochemical methods of lipopolysaccharide interacting with functional proteins and aptamers, and the reliability and cost need to be improved. We can identify different strains of lipopolysaccharide and bacterial mixtures as low as 10 ng / mL through the silicon nitride solid-state pore single-molecule device platform, and also distinguish different subtypes of the same strain. At the same time, we verified the specific single molecule interaction ability of lipopolysaccharide with antibiotic PMB and DNA aptamer. Finally, we extended the method to the detection of bacterial endotoxin in human blood samples and environmental water samples, and preliminarily verified the applicability of the scheme, providing a highly sensitive and simple endotoxin monitoring method for clinical diagnosis and environmental water quality. (Single-Molecule Lipopolysaccharides Identification and the Interplay with Biomolecules via Nanopore Readout, Biosens. Bioelectron. 2023, 240, 115641)

The nucleolin protein is widely present in the human nucleus and controls the metabolism of nucleic acids and chromatin structure. In particular, it can selectively bind to DNA / RNA G-quadruplexes and participate in the biological function of miRNA and the regulation of cancer cell overexpression. We first identified the DNA / RNA G-quadruplex conformations of different oncogenes with a difference of one base length by solid-state single-molecule technology, and also realized the sensitive detection of 100 pM nucleolin protein and the specific binding ability of nucleolin to AS1411-G4 and pre-miRNA 92b, pre-miRNA 149. The selective binding of nucleolin to these oncogene structures was also confirmed in human blood samples. This work provides a new idea for studying the single-molecule conformational differences of oncogene quadruplex and the specific binding mechanism of protein and nucleic acid advanced structure, and also provides a reference for the diagnosis process of nucleolin protein-related diseases. (Single-molecule recognition of Nucleolin and the interactions with DNA/RNA G-quadruplexes via nanopore decoding, Chem. Eng. J., 2023,473, 145311)

C-reactive protein (CRP) is a marker of inflammatory response and is also closely related to the pathology and cancer progression of many diseases. The single-molecule solid-state pore platform also achieved CRP detection with a lower limit of 0.3 ng / μL, and confirmed the selective interaction between CRP and aptamers and the sensitive monitoring of CRP in simulated blood samples. (Single-molecule Identification of the Conformations of Human C-reactive Protein and Its Aptamer complex with Solid-State Nanopore, ACS Appl. Mater. Inter., 2022, 14, 10, 12077-12088)

The research group also systematically reviewed the design of nanopore carrier materials and the principle of single molecule sensing, and summarized and prospected the application prospect, new material development and detection principle innovation in the field of nanopore single molecule sensing. (Overview of the materials design and sensing strategies of Nanopore devices, Coor. Chem. Rev., 2023, 478, 214998)

The above work was funded by the Western Light of the Chinese Academy of Sciences, Chongqing Science and Technology Bureau.

Related paper links

https://doi.org/10.1016/j.bios.2023.115641;

https://doi.org/10.1016/j.cej.2023.145311;

https://doi.org/10.1021/acsami.2c00453;

https://doi.org/10.1016/j.ccr.2022.214998.