Validation of a Commercial ELISA Kit for Screening 3-amino-2-oxazolidinone, A Furazolidone Antibiotic Residue in Shrimp

Abstract

Furazolidone, a banned nitrofuran antibiotic, is still illegally used in aquaculture and livestock husbandry. A commercial Enzyme-linked Immunosorbent Assay (ELISA) from Europroxima, The Netherlands is described, and validated for the screening of 3-amino-2-oxazolidinone (AOZ), metabolite of furazolidone antibiotic in the muscle of shrimp. Research works was conducted at Veterinary Drug Residue Analysis Laboratory, Institute of Food and Radiation Biology, Atomic Energy Research Establishment, Savar, Dhaka-1349, between July 2018 and December 2019. Validation was done in accordance with the European Commission (EC) decision 2002/657/EC in terms of specificity, detection capability CCβ, selectivity, ruggedness, and stability. Homogenized sample was subjected to acid hydrolysis for releasing of tissue bound metabolite AOZ, and derivatized with o-nitrobenzaldehyde followed by ethyl acetate/n-hexane extraction and detection by ELISA reader. The detection capability CCβ of the method was lower or at 0.3 µg/kg accepting 0% false compliant result. Cut-off factor Fm and the positivity threshold value T was 0.716 and 0.982, respectively. Mean OD of blank sample B was1.225. The assay was valid because T was greater than Fm and smaller than B. The kit was stable even after ten months of storage at recommended temperature and showed good sensitivity. From the practical point of view, the kit studied has good reliability, and could be used in the routine application for the control of illegal use of the drug.