Simultaneous detection and quantification of antimalarial drugs is important for evaluating the spread of their resistance. On the base of the target-induced assembly of branched DNA nanostructure signal amplification and different electrochemical coding tags at distinct potentials, an aptasensing method for multiplexed and highly sensitive detection of piperaquine (PQ) and mefloquine (MQ) antimalarial drugs is established. The target drugs associate with and switch the conformation of the aptamers to release free ssDNA trigger sequences, which initiate the self-assembly of many methylene blue (MB)- and ferrocene (Fc)-labeled auxiliary signal sequences to form branched nano-assemblies on electrode. The electrode-captured MB and Fc tags thus yield highly magnified current reponses at well-resolved potentials for simultaneously monitoring PQ and MQ with respective detection limits as low as 0.061 ng mL −1 (i.e., 0.114 nM) and 0.17 ng mL −1 (i.e., 0.449 nM). Moreover, the simultaneous detection of PQ and MQ in diluted human serum samples has also been verified by the developed sensor, revealing its potential for multiplexed monitoring of different drugs in connection with the corresponding aptamers for various application purposes.
Malaria remains a significant threat to global health, especially with the emergence of drug-resistant strains. Therefore, precise monitoring of antimalarial drug levels is crucial for assessing and managing drug resistance. Here, we report a multiplex aptasensor that enables simultaneous fluorescence detection of two antimalarial drugs (MQ and PQ) via split dsT7 promoter-mediated transcription of two-color light-up RNA aptamers. The detection mechanism relies on specific binding of target antimalarial drugs (MQ/PQ) to their aptamers, each triggering formation of intact dsT7 promoter to induce amplified transcription of its cognate RNA aptamer. The generated RNA aptamers bind their corresponding organic dyes (TO1-Biotin and MG) with high affinity, yielding 7.7-/25-fold fluorescence enhancement to enable ultrasensitive detection of MQ and PQ with limits of 0.71 ng/mL and 0.069 ng/mL, respectively. The method also exhibits high selectivity and low background signal. Furthermore, this label-free system achieves multiplexed detection of antimalarial drugs in 10-fold diluted human serum (recovery: 98.67–106.84 % for PQ and 99.89–106.19 % for MQ), demonstrating its real utility. This label-free system is adaptable to other biomarkers through aptamer replacement.
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