An Ultrasensitive ELISA for Medroxyprogesterone Residues in Fish Tissues Based on a Structure-Specific Hapten
2015
Kong, Na | Guo, Lingling | Guan, Dandan | Liu, Liqiang | Guang, Hua | Xu, Chuanlai
Medroxyprogesterone (MP) is an anabolic steroid, and its use is forbidden in livestock production. To enforce this ban, it is necessary to develop a highly specific and accurate enzyme-linked immunosorbent assay (ELISA) to detect MP residue. Herein, we describe the first synthesis of a specific hapten which retains the specific structure of MP by replacing the hydroxy group of MP with bromoacetic acid. This MP hapten was conjugated with bovine serum albumin and used as the immunogen in ELISA. The monoclonal antibody was produced by cell fusion. An indirect competitive ELISA was developed based on the monoclonal antibody for the detection of MP, which had a half inhibition concentration of 0.06 ng/mL and a limit of detection of 0.01 ng/mL. No cross-reactivity was found with eight analogous compounds by this method, which showed high specificity for MP. Recoveries ranged from 81 to 88 %, indicating the suitability of this ELISA for MP analysis in food samples.
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