Multiplexed aptasensor based on metal ions labels for simultaneous detection of multiple antibiotic residues in milk

A dual-target electrochemical aptasensor was developed for the simultaneous detection of multiple antibiotics based on metal ions as signal tracers and nanocomposites as signal amplification strategy. Metal ions such as Cd²⁺ and Pb²⁺ could generate distinct differential pulse voltammetry (DPV) peaks. When targets were present, kanamycin (KAN) and streptomycin (STR) as models, the KAN aptamer (KAP) and STR aptamer (STP) were released from their complementary strands, with more change of Cd²⁺ and Pb²⁺ corresponding to peak currents. At the same time, complementary strand of KAP (cKAP) and STP (cSTP) were linked with the poly (A) structure (cSTP-PolyA-cKAP) to increase their conformational freedom. Graphitized multi-walled carbon nanotubes (MWCNT_Gr) and carbon nanofibers-gold nanoparticles (CNFs-AuNPs) as a biosensor platform enhanced the surface area to capture a large amount of cSTP-PolyA-cKAP, thus amplifying the detection response. Under the optimal conditions, the aptasensor could detect KAN and STR as low as 74.50 pM and 36.45 pM respectively with the range from 0.1 to 100 nM and exhibited excellent selectively. Moreover, this aptasensor showed promising applications for the detection of other analytes by changing corresponding aptamers.