Validation of a Five Plate Test, the STAR protocol, for the screening of antibiotic residues in muscle from different animal species according to the European decision 2002/657/EC

Valerie Gaudin (1) , Celine Hedou (2) , Annie Rault (2) , Eric Verdon (2)

1 LERMVD
2 LERMVD

The STAR protocol is a Five Plate Test (FPT) developed several years ago at the Community Reference Laboratory (CRL) for the screening of antimicrobial residues in milk and muscle. This paper presents the validation of this method according to the European Decision 2002/657/EC and to an internal guideline for validation. A validation protocol based on "simulated tissues" and on a list of 16 representative antimicrobials to be validated has been implemented in our laboratory during several months for the STAR protocol. The performance characteristics of the method have been determined (specificity, detection capabilities CCβ, applicability, ruggedness). In conclusion, the STAR protocol is applicable to a broad spectrum detection of antibiotic residues in muscles of different animal species (pig, cattle, sheep, poultry). The method has good specificity (false positive rate 4%). The detection capabilities have been determined for 16 antibiotics from different families in relation to their respective Maximum Residue Limit (MRL): beta-lactams (penicillins and cephalosporins ≤ MRL), tetracyclines (≤MRL and ≤ 2.5 MRL), macrolides (2 MRL), quinolones (≤ 2 MRL), some sulphonamides (≤ 3 MRL), trimethoprim (2 MRL). However, the sensitivity of the STAR protocol towards aminoglycosides (> 8 MRL) and florfenicol (≤ 10 MRL) was unsatisfactory (>> MRL). The 2 objectives of this study have been met: first validate the STAR protocol according to the European decision 2002/657/EC, then demonstrate that the validation guideline (Anonymous 2010) developed to implement this decision is applicable to microbiological plate tests even for muscle. The use of "simulated tissue appeared as a good compromise between spiked discs with antibiotic solutions and incurred tissues. In addition, the choice of a list of representative antibiotics allowed to reduce the scope of the validation, which was already costly in time and effort.

Mots clés: Life Sciences

Valerie Gaudin, Celine Hedou, Annie Rault, Eric Verdon. Validation of a Five Plate Test, the STAR protocol, for the screening of antibiotic residues in muscle from different animal species according to the European decision 2002/657/EC. Food Additives and Contaminants, 2010, 27 (07), pp.935-952. ⟨10.1080/19440041003677483⟩. ⟨hal-00598938⟩