期刊
SENSORS AND ACTUATORS B-CHEMICAL
卷 387, 期 -, 页码 -出版社
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2023.133815
关键词
Myeloperoxidase; Phosphorothioate-modified DNA; DNAzyme; Terminal deoxynucleotidyl transferase; Electrochemical biosensor
Using a new phosphorothioate-modified DNA hairpin probe, we developed a label-free and highly sensitive electrochemical sensor for detecting abnormal myeloperoxidase (MPO) expression associated with inflammation and neurodegenerative diseases. This sensor combines DNAzyme-assisted cleavage recycling with DNA polymerization reaction to monitor MPO. It exhibits a linear range of 0.1-100 ng mL-1 and achieves a low detection limit of 0.046 ng mL-1 for MPO. The sensor is also highly selective against interferents and capable of detecting MPO in diluted human serum samples.
Abnormal myeloperoxidase (MPO) expression is associated with inflammation and neurodegenerative diseases. By using a new phosphorothioate (PT)-modified DNA hairpin probe, we fabricate a label-free and highly sensitive electrochemical sensor for monitoring MPO via integration of DNAzyme-assisted cleavage recycling with DNA polymerization reaction catalyzed by terminal deoxynucleotidyl transferase (TdT). The target MPO mediates the specific cleavage of PT-modified hairpin probes with assistance of hydrogen peroxide and chloride ion to initiate DNAzyme-based cyclic cleavage of substrate hairpin probes for the release of numerous trigger strands. Subsequent hybridizations of the trigger strands with the thiolated-hairpin probes on the electrode surface expose the 3 '-OH termini of hairpin probes for triggering TdT-catalyzed in situ DNA polymerization formation of long ssDNAs containing multiple G-quadruplex repeats. Hemin is confined on the electrode surface via the formation of G-quadruplex/hemin complexes to produce highly enhanced current signals for the determination of MPO in a sensitive and label-free way. The proposed sensor exhibits a linear range of 0.1-100 ng mL-1 and achieves a low detection limit of 0.046 ng mL-1 for MPO. Such sensor also shows high selectivity against other interferents and shows the capability to detect MPO in diluted human serum samples.
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