Saint-André-de-la-Marche, France
Saint-André-de-la-Marche, France

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Gaudin V.,Community Reference Laboratory | Hedou C.,Community Reference Laboratory | Rault A.,Community Reference Laboratory | Verdon E.,Community Reference Laboratory
Food Additives and Contaminants - Part A Chemistry, Analysis, Control, Exposure and Risk Assessment | Year: 2010

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 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 was implemented in our laboratory during several months for the STAR protocol. The performance characteristics of the method were determined (specificity, detection capabilities CC, applicability, ruggedness). In conclusion, the STAR protocol is applicable to the broad-spectrum detection of antibiotic residues in muscles of different animal species (pig, cattle, sheep, poultry). The method has good specificity (false-positive rate 1/44%). The detection capabilities were 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), and trimethoprim (2 MRL). However, the sensitivity of the STAR protocol towards aminoglycosides (48 MRL) and florfenicol (10 MRL) was unsatisfactory (44MRL). The two objectives of this study were met: firstly, to validate the STAR protocol according to European Decision 2002/657/EC, then to demonstrate that the validation guideline developed to implement this decision is applicable to microbiological plate tests even for muscle. The use of simulated tissue appeared a good compromise between spiked discs with antibiotic solutions and incurred tissues. In addition, the choice of a list of representative antibiotics allowed the reduction of the scope of the validation, which was already costly in time and effort. © 2010 Taylor & Francis.


Gaudin V.,Community Reference Laboratory | De Courville A.,Community Reference Laboratory | Hedou C.,Community Reference Laboratory | Rault A.,Community Reference Laboratory | And 3 more authors.
Food Additives and Contaminants - Part A Chemistry, Analysis, Control, Exposure and Risk Assessment | Year: 2013

Two microbiological kits based on Bacillus stearothermophilus (Eclipse 50® and Premi®Test) have been evaluated and validated according to the European guideline for the validation of screening methods (January 2010) and in relation to the concentrations recommended by the EU-RL in 2007. Both tests are robust, a fast method and easy to implement. Both tests are applicable to a very large variety of honeys from different floral and geographical origins (rosemary, lavender, scrub, heath, alder, forest, lemon, acacia, chestnut, raspberry, mountain and flowers) as well as honey of different colours (from blank honey to brown honey, including yellow and orange honey). A satisfactory false-positive rate of 5% was obtained for the Eclipse 50® test. The observed detection capabilities CCβ of the Eclipse 50® kit were: chlortetracycline (<75 μg kg-1), oxytetracycline (≤200 μg kg-1), tetracycline (<100 μg kg-1), cloxacillin (≤40 μg kg-1), tylosin (≤200 μg kg-1), desmycosin (<400 μg kg-1), sulfadiazine (≤300 μg kg-1), sulfadimethoxine (≤250 μg kg-1), sulfamerazine (<300 μg kg-1), sulfamethazine (<1000 μg kg-1), sulfamethizole (<75 μg kg-1), sulfamethoxazole (≤25 μg kg-1), sulfanilamide (≫1000 μg kg-1), sulfaquinoxaline (<75 μg kg-1), sulfathiazole (≤250 μg kg-1) and lincomycin (<1500 μg kg-1). These levels were all higher than the recommended concentrations where they exist. Due to its lack of sensitivity, it cannot be recommended for reliable routine use. The observed CCβ of the Premi®Test kit were: chlortetracycline (10 μg kg-1), oxytetracycline (<10 μg kg-1), tetracycline (≤10 μg kg-1), cloxacillin (≤5 μg kg-1), tylosin (≤10 μg kg-1), desmycosin (≤15 μg kg-1), sulfadiazine (≤25 μg kg-1), sulfadimethoxine (≤25 μg kg-1), sulfamerazine (≤25 μg kg-1), sulfamethazine (≤25 μg kg-1), sulfamethizole (≤25 μg kg-1), sulfamethoxazole (≤10 μg kg-1), sulfanilamide (≤25 μg kg-1), sulfaquinoxaline (≤10 μg kg-1), sulfathiazole (25 μg kg-1) and lincomycin (≤25 μg kg-1). The Premi®Test kit could be recommended for reliable use in routine control due to its low detection capabilities (except for aminoglycosides), but the disadvantage is a high false-positive rate of 14%. © 2013 Copyright Taylor and Francis Group, LLC.


PubMed | Community Reference Laboratory
Type: Evaluation Studies | Journal: Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment | Year: 2013

Two microbiological kits based on Bacillus stearothermophilus (Eclipse 50 and PremiTest) have been evaluated and validated according to the European guideline for the validation of screening methods (January 2010) and in relation to the concentrations recommended by the EU-RL in 2007. Both tests are robust, a fast method and easy to implement. Both tests are applicable to a very large variety of honeys from different floral and geographical origins (rosemary, lavender, scrub, heath, alder, forest, lemon, acacia, chestnut, raspberry, mountain and flowers) as well as honey of different colours (from blank honey to brown honey, including yellow and orange honey). A satisfactory false-positive rate of 5% was obtained for the Eclipse 50 test. The observed detection capabilities CC of the Eclipse 50 kit were: chlortetracycline (>75 g kg(-1)), oxytetracycline (200 g kg(-1)), tetracycline (>100 g kg(-1)), cloxacillin (40 g kg(-1)), tylosin (200 g kg(-1)), desmycosin (>400 g kg(-1)), sulfadiazine (300 g kg(-1)), sulfadimethoxine (250 g kg(-1)), sulfamerazine (>300 g kg(-1)), sulfamethazine (>1000 g kg(-1)), sulfamethizole (>75 g kg(-1)), sulfamethoxazole (25 g kg(-1)), sulfanilamide (>>1000 g kg(-1)), sulfaquinoxaline (>75 g kg(-1)), sulfathiazole (250 g kg(-1)) and lincomycin (>1500 g kg(-1)). These levels were all higher than the recommended concentrations where they exist. Due to its lack of sensitivity, it cannot be recommended for reliable routine use. The observed CC of the PremiTest kit were: chlortetracycline (10 g kg(-1)), oxytetracycline (>10 g kg(-1)), tetracycline (10 g kg(-1)), cloxacillin (5 g kg(-1)), tylosin (10 g kg(-1)), desmycosin (15 g kg(-1)), sulfadiazine (25 g kg(-1)), sulfadimethoxine (25 g kg(-1)), sulfamerazine (25 g kg(-1)), sulfamethazine (25 g kg(-1)), sulfamethizole (25 g kg(-1)), sulfamethoxazole (10 g kg(-1)), sulfanilamide (25 g kg(-1)), sulfaquinoxaline (10 g kg(-1)), sulfathiazole (25 g kg(-1)) and lincomycin (25 g kg(-1)). The PremiTest kit could be recommended for reliable use in routine control due to its low detection capabilities (except for aminoglycosides), but the disadvantage is a high false-positive rate of 14%.

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