A time-resolved fluorescence immunoassay for the ultrasensitive determination of diethylstilbestrol based on the double-codified gold nanoparticles

作者：Wang Longjun; Zhang Yuanfu; Liu Guofu; Zhang Chunyan; Wang Shuhao^*

来源：Steroids, 2014, 89: 41-46.

DOI：10.1016/j.steroids.2014.07.014

• Time-resolved fluorescence immunoassay
• Double-codified gold nanoparticles
• Amplification technique
• Diethylstilbestrol

An ultrasensitive and selective method is presented for the determination of diethylstilbestrol (DES) using time-resolved fluorescence immunoassay (TRFIA) based on double-codified gold nanoparticles (DC-AuNPs). In this system, the DC-AuNPs, that are gold nanoparticles (AuNPs) modified with anti-DES antibody and SH-dsDNA-biotin, was regarded as signal amplifier. A competitive immunoreaction was performed on polystyrene microtitration plates, where the DES compete with the immobilized DES-ovalbumin on polystyrene microtitration plates to bind to anti-DES antibodies on DC-AuNPs, and the europium(III)-labeled streptavidin was added to link to the SH-dsDNA-biotin as a tracer. Fluorescence signal was amplified via the AuNPs and the biotin-streptavidin double amplification systems. Under the optimized condition, DES can be quantified by TRFIA. The linear range and the limit of detection of DES were 1.0 x 10(-6)-10 ng mL(-1) and 0.4 fg mL(-1), respectively. This method was applied to determine DES in beef sample, with the recoveries ranging from 88% to 105%.

• 出版日期2014-11
• 单位聊城大学