Simultaneous Determination of Antibiotics, Mycotoxins, and Hormones in Milk by an 8–17 DNAzyme-Based Enzyme-Linked Immunosorbent Assay

Abstract

The simultaneous detection of three kinds of small-molecule contaminants (antibiotics, mycotoxins, and hormones) in milk was realized by using an 8–17 DNAzyme-based fluorescent enzyme-linked immunosorbent assay (ELISA), in which 8–17 DNAzyme was utilized as the catalytic enzyme for amplifying the signal. Compared with the conventional ELISA in which horseradish peroxidase is used as the catalyzing factor, this 8–17 DNAzyme-based ELISA could achieve multicolor signal output with lower detection limits. The linearities for chloramphenicol, 17β-estradiol, and aflatoxin M1 were in the range of 0.3 ng/mL–3 μg/mL, 3 ng/mL–3 μg/mL, and 3 pg/mL–3 ng/mL with quantitation limits of 0.3, 3, and 0.003 ng/mL, respectively. This proposed scheme demonstrated that the 8–17 DNAzyme might be an effective substitute for horseradish peroxidase in ELISA for the development of ultrasensitive and multicolor fluorescence immunoassay, which would stimulate the development of ELISA in a new orientation.

Aptamer‐Based Antibiotic Electrochemical Detection Platform Using Portable Plastic Gold Electrode and Multichannel Chip

“…Traditional detection techniques for antibiotic usually include methods based on chromatography/mass spectrometry, [ 5‐12 ] microbiological assay, [ 13‐15 ] and enzyme‐linked immunosorbent assay (ELISA). [ 16‐22 ] Due to the tedious and lack of portability of pre‐and post‐processing, chromatography/mass spectrometry methods are usually expensive and time‐consuming, while it is difficult to achieve repetitive analysis and accurate quantification in the microbiological assay. In comparison to the above two methods, although ELISA has been commercialized, it still has some problems such as enzyme inactivation and long detection time.…”

A Metasurface Plasmonic Analysis Platform Combined with Gold Nanoparticles for Ultrasensitive Quantitative Detection of Small Molecules

“…The excessive use of sulfamethazine can leave drug residues in food [5], and their residues in poultry eggs may cause health issues, such as allergic reactions, destruction of the hematopoietic system, and emergence of drug-resistant strains [5,6]. Currently, the methods most commonly used to monitor small molecules include liquid chromatography-mass spectrometry [7], enzyme-linked immunosorbent assay (ELISA) [8], and thin-layer chromatography (TLC) [9]. These methods can be expensive, use large and complex equipment that require skilled operation, are time-consuming, or have limited sensitivity [10,11], potentially preventing their broad application in point-of-care diagnostics and private laboratories.…”