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Direct enzyme immunoassay of progesterone in bovine plasma
Author(s) -
ISOBE Naoki,
NAKAO Toshihiko
Publication year - 2003
Publication title -
animal science journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.606
H-Index - 38
eISSN - 1740-0929
pISSN - 1344-3941
DOI - 10.1046/j.1344-3941.2003.00128.x
Subject(s) - chemistry , chromatography , immunoassay , horseradish peroxidase , peroxidase , assay sensitivity , enzyme , endocrinology , medicine , antibody , biochemistry , biology , immunology , alternative medicine , pathology
The present study was undertaken to develop a novel, practical and simple procedure for enzyme immunoassay (EIA) of plasma progesterone in cows. Diluted plasma was heated for 70°C for 30 min and applied directly to wells of a microtitre plate without extraction. Then plasma was incubated with antiprogesterone antibody and horseradish peroxidase‐labeled progesterone. The sensitivity of the assay was estimated as 4.4 pg/mL (0.11 pg/well). The intra‐assay and interassay coefficients of variation were 5.7–19.1% and 6.6–19.3%, respectively. When 0.3, 1 and 3 ng of progesterone were added to plasma, the recovery rates ranged between 79.9 and 108.4%. Only 4 h were needed to complete an assay to measure progesterone concentration. To apply the present direct EIA, progesterone concentration in plasma was assayed in crossbred cows used for the embryo transfer program. During insertion of controlled‐internal drug release (CIDR), progesterone concentrations were kept at a high level, although the removal of CIDR with treatment of dinoprost trometamine reduced progesterone concentration drastically. These results suggest that the present direct EIA is a practical and suitable method for measuring the plasma concentration of progesterone.