Establishment of a dual-aptasensor for simultaneous detection of chloramphenicol and kanamycin

Abstract

ABSTRACT 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.