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1993, Number 3

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Vet Mex 1993; 24 (3)

Antibodies obtention against progesterone and estradiol, radioimmunoassay standardization and validation in ruminant serum

Herrera DMR, Luna MM, Romero RCM

Language: English/Spanish
References: 37
Page: 223-230
PDF size: 164.01 Kb.


ABSTRACT

Male rabbits were weekly immunized six times with; 100 µg of progesterone-CMO-BSA or with 160 µg of estradiol-17ß in order to obtain specific antibodies against progesterone (P4) and estradiol-17ß (E2) as well as the standardization of radioimmunoassays (RIASs). Titration was made weekly and the best titrer was chosen for cross reaction with other steroids. Results of the cross reaction for antisera against P4 were: 4.4 and 4.4% for pregnenolone; 4.0 and 13.3% for epipregnenolone and ‹ 1.0% for alopregnenolone, testosterone, E2 and cortisol for AcQKP5 and AcQKP6, respectively. Results of cross reaction of antisera against E2 were: 3.6% for estrone and ‹ 0.01% for estriol, androstendione, estradiol-17α, P4 and testosterone. Assayssensivity were 6.25 pg Of P4 or 0.5 pg of E2. Intra-and inter-assay coefficients of variation were: 10.9% and 12.8% for P4, and 5.0% and 10.0% for E2, respectively. RIAs validation was achieved by determining P4 and E2, plasma concentration in goats and cows during different reproductive stages. Results obtained for P4 in goats were 8.3 ± 0.4 ng/ml during the last third of gestation. During labour period, P4 concentrations decreased to 5.4 ± 0.8 ng/ml. E2 values in labor period were 55.1 ± 8.1 pg/ml. Values obtained in cows for P4 and E2 were: 0.9 ± 0.08 ng/ml and 10.6 ± 2.4 pg/ml during estrous; 8.6 ± 2.7 ng/ml and 3.3±0.6 pg/ml during luteal phase; and 6.4 ± 0.6 ng/ml and 17.3 ± 1.6 pg/ml during gestation, respectively. These results show that RIAs sensivity with the obtained antibodies is suitable for detecting typical values of P4 and E2 in domestic ruminants during different reproductive stages.


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