Foodborne bacterial infections pose serious threats to human health, leading to illnesses and even fatalities. The key to solve this problem is to develop fast and effective sensor for sensitive detection of bacteria. In this study, an electrochemical biosensor based on magnetic nanocomposite material Fe 3 O 4 @COF-AuNPs and trigging isothermal circular amplification (TICA) was constructed for detection of foodborne bacteria. In this design, a hairpin aptamer probe (HAP)-bound DNA monolayer was designed, serving as the substrate for the electrode to increase surface area. The Escherichia coli ( E. coli ) was selected as the target. The presence of E. coli caused the configuration change of the HAP and allowed the occurrence of TICA with the use of the Klenow Fragment polymerase (KF-Poly), thus the HAPs were away from the DNA monolayer. After adding the catalytic magnetic nanocomposite material Fe 3 O 4 @COF-AuNPs, it was immobilized on the DNA monolayer of the electrode through DNA complementary pairing and exhibited high electrocatalytic behavior towards the reduction of 4-nitrophenol. This enabled ultrasensitive detection of E. coli , with the detection limit (LOD) as low as 10 CFU/mL. The detection time was 1 h. This designed electrochemical sensor showed high selectivity, stability, and reproducibility, which offered a new avenue for detection of foodborne bacteria.
Wei Yuxin,Fu Yang,Li Chuanhai,Chen Siyu,Xie Linzhi,Chen Mei
A electrochemical biosensor was designed utilizing a CRISPR Cas9n-driven DNA walker combined with gold-nanosphere-like covalent organic frameworks (COFs-AuNPs) to detect breast cancer markers (PIK3CA E545K ctDNA). The DNA walker probe is activated only in the presence of circulating tumor deoxyribonucleic acid (ctDNA), binding to a support probe to form a double strand that is then specifically cleaved by the Cas9n/sgRNA complex. This cleavage produces numerous DNA fragments for signal amplification. The COF-AuNPs as electrode materials facilitate electronic transfer and provide additional active sites for the immobilization of nucleic acid probes. This setup achieves a detection limit of 1.76 aM, demonstrating high sensitivity. Additionally, Cas9n improves the specificity of the sensor, accurately distinguishing a pair of base-mismatched sequences, and reducing the occurrence of false positives. Overall, the sensor exhibits excellent selectivity, reproducibility, and potential for early diagnosis of breast cancer. Graphical Abstract