Abstract
ABSTRACT An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on immunomagnetic beads' (IMBs) clean-up was developed for detection of the residues of maduramicin (MD) in different chicken tissues. IMBs coated with a specific monoclonal antibody (MAb) against MD named MAb 2D6 were applied to enrich the residues of MD in chicken tissues. The specificities of these IMBs were well maintained and the reversibility remained at more than 73% of the original capability after being used for three times. After elution, enriched MD was detected by a conventional ic-ELISA. The limits of detection of MD were 72, 74 and 173 μg/kg in chicken muscle, skin and fat, and liver, respectively. Recoveries ranged from 80.0% to 115.8% with coefficients of variation being less than 11.3%. These results indicated that a rapid, robust clean-up of IMBs combining ELISA provides a simple, time-saving and environmentally friendly method to detect MD in chicken tissues.
Highlights
Polyether antibiotics are widely used as feed additives for the prevention of coccidiosis in chickens (Stanker, Elissalde, Rowe, Beier, & El-Aziz Nasr, 1994; Tian et al, 2017)
In order to prepare immunomagnetic beads’ (IMBs), hydrophilic magnetic beads with carboxyl groups on the surface were used because carboxyl acid groups on the surface of Magnetic bead (MB) could be coupled with the amino groups in monoclonal antibody (MAb) by ethyl-3-(3-dimethylaminopropy) carbodiimide (EDC) and NHS (Jang & Keng, 2008; Song et al, 2014)
MAb 2D6 with lower affinity constant was selected to couple with MBs, and MAb 3B4 with higher affinity constant was chosen for Enzyme-linked immunosorbent assay (ELISA) analysis
Summary
Polyether antibiotics are widely used as feed additives for the prevention of coccidiosis in chickens (Stanker, Elissalde, Rowe, Beier, & El-Aziz Nasr, 1994; Tian et al, 2017). Maduramicin (MD), a polyether ionophore antibiotic, is extensively used to prevent coccidiosis in poultry due to its board anticoccidial activity. MD has been widely used as a growth promoter to improve feed conversion rate in ruminants (Le-Minh, Khan, Drewes, & Stuetz, 2010). The extensive or illegal use of MD can lead to intoxication of animals and residues in chicken tissues (Wang, Zhang, Murtazina, Eremin, & Shen, 2008), which constitutes a potential hazard for both human and environment’s health (Sharma, Bhalla, Varma, Jain, & Singh, 2005).
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