DETECTION AND QUANTIFICATION OF PROGESTER ONE IN SERUM SAMPLES USING ENZYME-LINKED IMMUNOASSAY (ELISA)

A method for the detection and quantification of progesterone residues in serum samples using enzyme linked immune sorbent assay (ELISA) is being presented. Furthermore, the method has been validated according to the criteria listed in the appendix of the European Community's gui deline 85/591/EWG. Sample preparation is performed by extracting the s erum with diethyl-ether, evaporation and takeup of the dry residue in buffer solution, the progesterone content of which is determined using a microtiter plate coated with the appropriate antibodies. The time r equired for 40 samples including the preparation of chemicals and appa ratus, measurement and computer-aided calculation of the results varie s within 5 and 5.5 hours. The quality criteria, upon which the validat ion of the presented method is based, have been derived by fortificati on experiments using the concepts of DIN 32645 and DFG (1991) for an e stimation of the limits of detection and of determination. The limit o f detection was calculated as 0.13 ng/ml, the limit of determination b eing 0.30 ng/ml, the method's sensitivy 0.85 and the method's variatio n coefficient 7.4%.