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Aptamers, as single-stranded DNA or RNA fragments, have been widely applied as the bio-recognition element for fabrication of flexible and reliable aptasensors to be used in food safety control, clinical therapy and diagnosis and environment monitoring fields. With increasingly fierce antibiotics resistance appearing as a worldwide problem, a highly efficient method is urgently needed to detect antibiotics residues in animal-sourced food. Herein, a simply operated aptasensor based on quantitative real-time PCR (qRT-PCR) was fabricated to realise the simultaneous detection of two antibiotics (i.e. chloramphenicol and kanamycin). The limit of detection (LOD) of 6.13 ng/mL for chloramphenicol and of 19.17 ng/mL for kanamycin of this dual-aptasensor were achieved. Actually, such LOD values were not as good as that of an aptasensor individually established for each antibiotic. The circular dichroism analysis suggested that in the dual-aptasensor, adjacent aptamers might disturb each other's binding with their respective target. Although certain detection sensitivity was lost, the dual-aptasensor could still fulfil the detection requirements, and more importantly, it would improve the detection efficiency. Finally, this dual-aptasensor was applied for detecting chloramphenicol and kanamycin in real spiked food samples, and results indicated good recovery rates. These results demonstrated this developed dual-aptasensor to be a promising highly efficient method with low cost for simultaneous detection of chloramphenicol and kanamycin residues in animal-sourced food samples. [ABSTRACT FROM AUTHOR]

[Display omitted] • Study on the mechanism of aptamer-based fluorescent DNA biosensors. • Antibiotic detection is facilitated by the construction of aptamer-based fluorescent DNA biosensors using both label-free and labeled methods. • Signal amplification are harnessed to pursuit the heightened sensitivity of the antibiotic detection. The inappropriate employment of antibiotics across diverse industries has engendered profound apprehensions concerning their cumulative presence within human bodies and food commodities. Consequently, many nations have instituted stringent measures limiting the admissible quantities of antibiotics in food items. Nonetheless, conventional techniques employed for antibiotic detection prove protracted and laborious, prompting a dire necessity for facile, expeditious, and uncomplicated detection methodologies. In this regard, aptamer-based fluorescent DNA biosensors (AFBs) have emerged as a sanguine panacea to surmount the limitations of traditional detection modalities. These ingenious biosensors harness the binding prowess of aptamers, singular strands of DNA/RNA, to selectively adhere to specific target antibiotics. Notably, the AFBs demonstrate unparalleled selectivity, affinity, and sensitivity in detecting antibiotics. This comprehensive review meticulously expounds upon the strides achieved in AFBs for antibiotic detection, particularly emphasizing the labeling modality and the innovative free-label approach. It also elucidates the design principles behind a diverse array of AFBs. Additionally, a succinct survey of signal amplification strategies deployed within these biosensors is provided. The central objective of this review is to apprise researchers from diverse disciplines of the contemporary trends in AFBs for antibiotic detection. By doing so, it aspires to instigate a concerted endeavor toward the development of heightened sensitivity and pioneering AFBs, thereby contributing to the perpetual advancement of antibiotic detection methodologies. [ABSTRACT FROM AUTHOR]