Cascaded strand displacement for non-enzymatic target recycling amplification electrochemical detection of SARS-CoV-2EI北大核心CSCD

An electrochemical biosensor for low cost and highly sensitive and selective detection of SARS-CoV-2 target nucleic acid was developed based on two cascaded toehold-mediated strand displacement reactions (TSDRs). Driven by thermodynamic entropy, the target nucleic acid bound to the first toehold region of the probes, leading to the first TSDR and the second toehold region exposed. Subsequently, the methylene blue (MB)-modified signal probe triggered the second TSDR and led to cyclic reuse of the target nucleic acid. Based on cascaded TSDRs, a large number of signal probes were combined on the sensor surface to produce significantly enhanced square wave voltammetry (SWV)electrochemical signals. The results showed that the SWV signal intensity was proportional to the logarithm of the target nucleic acid concentration, and had a good linear relationship in the range of 5×10-14-5×10-10 mol/L with a detection limit of 1.8×10-14 mol/L. Moreover, the sensor could be employed to monitor SARS-CoV-2 nucleic acid in 10% serum samples. © 2023, Youke Publishing Co.,Ltd. All rights reserved.     

Portable biosensor for quantitative detection of melamine based on CRISPRCas12a and a personal glucose meterEI北大核心CSCD

A CRISPR-Cas system holds great promise as a next-generation biosensing technology for molecular diagnostics. In this paper, a portable biosensor based on the trans-cleavage activity of CRISPR-Cas12a and a personal glucose meter has been developed for quantitative, sensitive and specific detection of melamine. The presence of the target melamine binds to the aptamer, leading to the release of locker DNA. And then, the leasing locker DNA activates the trans-cleavage activity of CRISPR-Cas12a to cleave the single-strand DNA (ssDNA) linker on sucrase-ssDNA modified electrode, releasing a short DNA fragment labeled with sucrase in the resulting solution. The sucrase could further catalyze sucrose to glucose, which could be detected by the PGM. Under the optimized conditions, the increase of PGM signal was relative with the concentration of melamine ranging from 0.1 to 2.5 μmol/L and the limit of detection (LOD) was 37 nmol/L. Moreover, the portable biosensor has strong specificity and can be used for the quantitative detection of melamine in milk samples. © 2023, Youke Publishing Co.,Ltd. All rights reserved.