An Ultrasensitive ELISA for Medroxyprogesterone Residues in Fish Tissues Based on a Structure-Specific Hapten
- PDF / 671,133 Bytes
- 8 Pages / 595.276 x 790.866 pts Page_size
- 77 Downloads / 186 Views
An Ultrasensitive ELISA for Medroxyprogesterone Residues in Fish Tissues Based on a Structure-Specific Hapten Na Kong & Lingling Guo & Dandan Guan & Liqiang Liu & Hua Kuang & Chuanlai Xu
Received: 20 August 2014 / Accepted: 7 October 2014 # Springer Science+Business Media New York 2014
Abstract 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. Keywords Medroxyprogesterone . Antigen . Monoclonal antibody . ELISA
Introduction Medroxyprogesterone (MP) is a type of anabolic steroid, which is used as therapy for dysmenorrhea, functional amenorrhea, and habitual abortion. Medroxyprogesterone can also be used in large doses as a long-term contraceptive (Leider 1981). In livestock farming, it is mainly used as a growth promoter and as a treatment for reproductive disease (Chan N. Kong : L. Guo : D. Guan : L. Liu : H. Kuang : C. Xu (*) State Key Lab of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi 214122, Jiangsu, People’s Republic of China e-mail: [email protected]
et al. 2014). However, potentially harmful effects have been reported (Barel-Cohen et al. 2006; Stanczyk and Bhavnani 2014) to be caused by the abuse of anabolic steroids in food and the environment. Thus, many countries including China have banned the addition of anabolic steroids to animal feed. Over the years, many techniques have been developed for the detection of anabolic steroid residues, such as high-performance liquid chromatography (HPLC) (Chanthai and Tessiri 2012), liquid chromatography (LC) coupled with mass spectrometry (MS) (Porphyre et al. 2013), solid-phase dispersion extraction (Yang et al. 2008), and gas chromatography (GC)-MS (Lu et al. 2013). However, the limitations of these techniques include their time-consuming and complexity of sample preparation. In order to avoid these disadvantages, more and more people are developing immunoassays to detect steroid residues (Liu et al. 2009; Peng et al. 2007). Immunoassays, especially enzyme-linked immunosorbent assay (ELISA), are a rapid and simple detection method based on antigen-antibody recognition (García-Cordero et al. 2014; Hua et al.
Data Loading...
