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

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, 17 beta-estradiol, and aflatoxin M1 were in the range of 0.3 ng/mL-3 mu g/mL, 3 ng/mL-3 mu g/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.

Automated summary

The study utilizes 8-17 DNAzyme as a substitute for horseradish peroxidase for simultaneous detection of small-molecule contaminants in milk, achieving multicolor signal output with lower detection limits. This approach may stimulate the development of ELISA in a new direction.