Colorimetric aptasensor for on-site detection of oxytetracycline antibiotic in milk

Oxytetracycline (OTC), one of the largely used antibiotic in veterinary practice has been banned due to its potential side effects. Development of a field applicable and affordable kit to detect OTC will help to eliminate such milk from human consumption. An aptamer has been designed (27 nt; K_d = 29.2 ± 19.4 nM) through rational truncation. OTC interacts with this aptamer in G rich regions as confirmed by molecular modelling and circular dichroism spectroscopy. To develop a lateral flow based aptasensor, OTC was conjugated with a 7 kDa carrier protein to immobilize onto the nitrocellulose membrane. Using 0.125 µM aptamer-gold conjugate, assay could visually detects upto 5 ng/mL of OTC in spiked milk within 10 mins [Limit of quantitation (LOQ)-0.254 ± 1.62 ng/mL; permissible limit 100 ng/mL]. It showed no cross reactivity with components of milk and data correlated with analysis done through HPLC.