Upconversion luminescence-based aptasensor for the detection of thyroid-stimulating hormone in serum

Abstract

Thyroid-stimulating hormone (TSH) plays a crucial physiological and pathological role in humans, and a timely and sensitive detection of TSH is critical for early diagnosis and prevention of thyroid-related diseases. Herein, we developed a simple wash-free biological aptasensor based on luminescence resonance energy transfer (LRET) between NaYF₄:Yb,Er upconversion nanoparticles (UCNPs) and tetramethylrhodamine (TAMRA) for the detection of TSH with high sensitivity. In this LRET system, UCNPs as donors and TAMRA as receptors were modified with nucleic acid aptamers Apt-1 and Apt-2, respectively. When TSH was present, the two aptamer strands both specifically recognized TSH to form a hairpin-like structure, thereby shortening the space between UCNPs and TAMRA. The LRET occurred under radiation of 980-nm light. By detecting the change of upconversion luminescence (UCL) intensity (I_545nm), the activity of TSH was quantified. The resulting detection dynamic range and the limit of detection were 0.1-5.0 mIU·L^-1 and 0.065 mIU·L^-1, respectively. The aptasensor using UCNPs as LRET donors was capable of effectively eliminating the background interference of a complicated biological environment, and showed good specificity because of the excellent recognition function of aptamers. Due to high sensitivity, easiness of fabrication, operational convenience, and selectivity, the UCL-based aptasensor is a promising candidate for clinical TSH determination. Based on nucleic acid aptamer and the mechanism of luminescence resonance energy transfer (LRET) between upconversion nanoparticles (UCNPs) donor and tetramethylrhodamine (TAMRA) receptor, an aptasensor was constructed for the quantitative analysis of TSH activity in serum by testing the change of I_545nm.

Keywords: Aptasensor; LRET; Tetramethylrhodamine; Thyroid-stimulating hormone; UCNPs.