Zhang Tao, Liu Guiling, Sun Siwei, Meng Zongwu, Qiu Yuzhe, Ding Ping
Xiangya School of Public Health, Central South University, Changsha, Hunan, 410078, China.
Hunan Provincial Key Laboratory of Clinical Epidemiology, Changsha, Hunan, 410078, PR China.
J Nanobiotechnology. 2025 May 14;23(1):346. doi: 10.1186/s12951-025-03454-3.
The antibiotic residues pose significant risks for bacterial resistance. To address the practical requirements for rapid, accurate, and on-site detection of antibiotic residues and monitoring the abundance of associated resistance genes, we report a smartphone-integrated multi-mode platform. The platform is aimed to simultaneous, accurate, and visual quantitative detection of ampicillin (AMP) and β-lactam antibiotic resistance genes (blaTEM). Specifically, we developed a magnetically controlled fluorescence, colorimetric, and photothermal biosensor based on a magnetic separation unit (aminated modified complementary DNA chain (NH-cDNA) loading on the surface of Ferrosoferric Oxide@polydopamine (FeO@PDA, FP), FP@cDNA) and a signal unit (the aptamer nucleic acid chain modified by phosphate group linked to Prussian blue@UiO-66@manganese dioxide (PB@UiO-66@MnO, PUM) through Zr-O-P bond, PUM@Apt), for the integrated detection of AMP and blaTEM. By utilizing complementary base pairing between FP@cDNA and PUM@Apt, along with precise aptamer recognition the AMP, we achieved the fluorescence quantitative detection of AMP by measuring the signal unit in the supernatant. Subsequently, the difference of signal units in colorimetric process leads to a varying conversion rate of oxidized 3,3',5,5'-Tetramethylbenzidine (oxTMB), enabling the output of colorimetric and photothermal signals. The competitive binding of aptamers permitting the determination of AMP in the range of 0-160 pM with a low detection limit (0.34 pM). Additionally, in the presence of blaTEM, the activated CRISPR/Cas12a indiscriminately cleaves the single-stranded portion of the FP@DNA@PUM complex obtained by magnetic separation. A PUM-based three-signal detection scheme was established for the sensitive determination of blaTEM with the limit of detection (LOD) of 1.03 pM. The integration of smartphone-assisted analysis broadens the potential of the platform for visual detection. Notably, the innovative platform, with its excellent stability, exhibits great potential as a simple yet robust approach for the simultaneously visually monitoring antibiotics and drug resistance genes, and holds promise in the field of kit development.
抗生素残留对细菌耐药性构成重大风险。为满足对抗生素残留进行快速、准确和现场检测以及监测相关耐药基因丰度的实际需求，我们报告了一种集成智能手机的多模式平台。该平台旨在同时、准确且可视化地定量检测氨苄青霉素（AMP）和β-内酰胺抗生素耐药基因（blaTEM）。具体而言，我们基于磁分离单元（负载在四氧化三铁@聚多巴胺（FeO@PDA，FP）表面的氨基化修饰互补DNA链（NH-cDNA），FP@cDNA）和信号单元（通过Zr-O-P键与普鲁士蓝@UiO-66@二氧化锰（PB@UiO-66@MnO，PUM）连接的磷酸基团修饰的适体核酸链，PUM@Apt）开发了一种磁控荧光、比色和光热生物传感器，用于AMP和blaTEM的集成检测。通过利用FP@cDNA与PUM@Apt之间的互补碱基配对，以及适体对AMP的精确识别，我们通过测量上清液中的信号单元实现了AMP的荧光定量检测。随后，比色过程中信号单元的差异导致氧化3,3',5,5'-四甲基联苯胺（oxTMB）的转化率变化，从而实现比色和光热信号的输出。适体的竞争性结合使得能够在0 - 160 pM范围内测定AMP，检测限低至0.34 pM。此外，在存在blaTEM的情况下，活化的CRISPR/Cas12a会无差别地切割通过磁分离获得的FP@DNA@PUM复合物的单链部分。建立了基于PUM的三信号检测方案，用于灵敏测定blaTEM，检测限（LOD）为1.03 pM。智能手机辅助分析的集成拓宽了该平台用于可视化检测的潜力。值得注意的是，这个创新平台具有出色的稳定性，作为一种简单而强大的同时可视化监测抗生素和耐药基因的方法具有巨大潜力，并在试剂盒开发领域有广阔前景。
