Picomolar chloramphenicol detection by orchestrating multi-level signal systems of metal-organic frameworks-catalytic hairpin assembly-gold nanoprobes

Chloramphenicol (CAP), a broad-spectrum antibiotic, faces stringent regulatory restrictions in clinical and food safety applications due to its severe toxicity. In practical scenarios, achieving highly sensitive and rapid detection of ultralow-level CAP presents a significant challenge in sensor system design. Here, we propose an orchestrated multi-level signal strategy to boost the sensitivity of an electrochemical aptasensor, enabling picomolar-level CAP detection. Amino-functionalized cobalt-based metal-organic framework nanosheets (Co-BDC(NH₂)) and thionine-labeled gold nanoparticles (thio-AuNPs) are synthesized as electron transfer carriers and signal amplifiers, respectively. In the presence of picomolar concentrations of CAP, the catalytic hairpin assembly (CHA) reaction of DNA is triggered, linking the aptamer-modified Co-BDC(NH₂) nanosheets with thio-AuNPs and facilitating precise CAP detection at low concentrations. Under optimal conditions, the aptasensor delivers an ultralow limit of detection (LOD) of 3.33 pM with a wide linear range from 10 pM to 50 nM. Notably, the aptasensor also exhibits excellent anti-interference capability, reproducibility, and stability in real matrices like milk, bottled water, and lake water. This work presents a high-sensitivity sensor platform design strategy that enables effective detection of ultralow-level targets beyond environmental substances through multiple signal amplification process.