Validation of a dissociation enhanced lanthanide fluorescence immunoassay for the screening of 17beta-estradiol in bovine serum according to European Union decision 2002/657/EC.
A validation study was carried out in order to evaluate the performances of a dissociation enhanced lanthanide fluorescence immunoassay (DELFIA) for rapid screening of 17 beta-estradiol in bovine serum. This validation was performed according to European Union (EU) Decision 2002/657/EC, which establishes criteria and procedures for determination of detection capability (CCbeta), selectivity/specificity, and applicability/ruggedness/stability for qualitative screening tests. To determine these performance characteristics, 20 blank serum samples of cattle were collected and spiked with 17 beta-estradiol at 40 pg/mL, corresponding to the maximum residue limit permitted by Italian legislation. According to the EU Decision CCbeta criterion, spiked samples must have <5% probability to be classified as a false negative. 17 beta-Estradiol was detected in each spiked sample, and the CCbeta results were <40 pg/mL. There was also no observed interference effect due to chemically related substances or from the matrix. Moreover, slight variations of some critical factors in the DELFIA procedure, deliberately introduced for ruggedness evaluation, did not result in any negative effect on the 17beta-estradiol detection. The proposed method is suitable for qualitative screening analysis of 17 beta-estradiol according to EU performance requirements.
Scalas
Daniela
D
Institute Zooprofilattico Sperimentale del Piemonte, Liguria eValle d'Aosta I. Altara, Via Bologna, 148, 10154 Torino, Italy.
Squadrone
Stefania
S
Gili
Marilena
M
Marchis
Daniela
D
Prearo
Marino
M
Abete
Maria Cesarina
MC
D000818:Animals; D002138:Calibration; D002417:Cattle; D002623:Chemistry Techniques, Analytical / Q000379:methods / Q000592:standards*; D004350:Drug Residues / Q000032:analysis; D004958:Estradiol / Q000032:analysis*; D005062:European Union; D005504:Food Analysis; D007118:Immunoassay / Q000379:methods*; D028581:Lanthanoid Series Elements / Q000032:analysis*; D015233:Models, Statistical; D011336:Probability; D012015:Reference Standards; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*
D028581:Lanthanoid Series Elements; D004958:Estradiol