A fluorescent sensing strategy for ultrasensitive detection of oxytetracycline in milk based on aptamer-magnetic bead conjugate, complementary strand of aptamer and PicoGreen

Misuse of antibiotics in animal husbandry and presence of their residues in animal foods isa serious crisis worldwide and thus, monitoring the level of them in food samples is vital for human health. Herein, a fluorescent aptasensor was developed for highly sensitive quantification of oxytetracycline (OTC) in food samples. This method is based on OTC aptamer conjugated to magnetic beads, functioned as recognition element, complementary strand of OTC aptamer, and PicoGreen (PG) as a sensitive double-stranded DNA (dsDNA) fluorescent dye. Formation of OTC aptamer-magnetic bead conjugate provides the opportunity of sample condensation and separation technology. Additionally, the presence of complementary strand leads to significant fluorescence signal alteration of aptasensor in the presence or absence of target and a noteworthy improvement of the aptasensor sensitivity. In the absence of target, complementary strand could bind to aptamer and form dsDNA on the surface of magnetic bead. As a consequence, adding PG to the sample leads to observation of high fluorescence signal from sample. In contrast, once OTC is added to the sample, it binds to OTC aptamer-magnetic bead complex and prevents hybridization of OTC aptamer and its complementary strand. Hence, after addition of PG to the sample, a weak fluorescence intensity is measured. Under optimized conditions, the linear ranges for OTC detection were 0.2-2 nM and 2-800 nM. The detection limit was calculated to be as low as 0.15 nM for the fabricated aptasensor. Besides the great sensitivity, proposed method demonstrated superior specificity towards OTC once it was used against several antibiotics. More significantly, the recovery rates of OTC in milk ranged from 96.46% to 101.5%, implying the great feasibility of designed sensoras well as its potential to be employed for analysis of OTC in real samples. (c) 2020 Elsevier B.V. All rights reserved.

Automated summary

A fluorescent aptasensor was developed for highly sensitive quantification of oxytetracycline in food samples. The method utilizes OTC aptamer conjugated to magnetic beads for sample separation, and the presence of a complementary strand significantly alters the fluorescent signal of the aptasensor, resulting in improved sensitivity. This approach demonstrated great feasibility and potential for analysis of OTC in real samples.