A signal-on fluorescent aptasensor by sensitized Tb3+ luminescence for detection of melamine in milk

Abstract

A fluorescent aptasensor based on sensitized terbium(III) luminescence was constructed to detect melamine in milk. Tb³⁺ as the fluorescence probe can be sensitized by a guanine-rich single-stranded DNA sequence, so the complementary sequence of the polythymidine aptamer (cDNA) was modified with six consecutive guanine bases (G6). In the absence of melamine, melamine aptamer combined with cDNA to form a double helix structure, and G6 hybridized with the extended cytosine bases in the aptamer, resulting in low fluorescence intensity of Tb³⁺. In the presence of melamine, cDNA was released due to the specific recognition of melamine to the aptamer, resulting in stronger sensitized fluorescence intensity of Tb³⁺. Under the optimum conditions, the linear concentration of melamine in the milk ranged from 1.0 μg/mL to 10.0 μg/mL. This aptasensor can be used for the accurate and rapid detection of melamine in milk with a detection limit of 0.02 μg/mL, and has the advantages of high sensitivity, high efficiency, simple operation and low cost.

Keywords: Aptamer; DNA-sensitized luminescence; Melamine; Terbium.