Articles

Coupling steroids with alkaline phosphatase for competitive immunoenzymatic assays in the plasma of various domestic mammals

Abstract

The concentrations of various steroids involved in animal reproduction and behavior, in particular progesterone, are analyzed by assays in the Endocrinology and Phenotyping Laboratory. Since progesterone is known to play a major role in reproduction in females, its concentration in plasma is analyzed by competitive immuno-enzymatic assay. With the halting of the sale of progesterone coupled with alkaline phosphatase (P4-Pal), the reactant essential for this competitive assay, we decided to develop a protocol for coupling steroids with alkaline phosphatase. Following this coupling for progesterone, and in the framework of the laboratory’s quality approach, the reliability, repeatability and reproducibility of the results were assessed. Since all these quality standards were satisfactorily met, we looked for the existence of correlations between the concentrations obtained in plasma samples with assays using the P4-Pal produced in the laboratory and those obtained using the P4-Pal initially marketed. For the 5 animal species studied mostly in the laboratory, the correlation coefficients r varied between 0.92 and 0.98. The protocol for coupling progesterone with alkaline phosphatase has been validated.

References

  • Canépa S, Lainé AL, Bluteau A, Fagu C, Flon C, Monniaux D (2008), Validation d'une méthode immunoenzymatique pour le dosage de la progestérone dans le plasma des ovins et des bovins. Cah. Techn. Inra, 64, 19-30.
  • Driancourt MA, Freret S, Saint Dizier M (2014), Les cycles œstriens. La reproduction animale et humaine, Saint Dizier M., Chastant-Maillard S., coord, Eds, Quae, Versailles Chapitre 12, 219-234.
  • Saumande J, Tamboura D and Chupin D (1985), Changes in milk and plasma concentrations of progesterone in cows after treatment to induce superovulation and their relationships with number of ovulations and of embryos collected. Theriogenology 23, 719-731. https://doi.org/10.1016/0093-691X(85)90147-5
  • Sesay AM, Micheli L, Tervo P, Palleschi G, Virtanen V (2013), Development of a competitive immunoassay for the determination of cortisol in human saliva. Anal. Biochem, 434, 308-314. https://doi.org/10.1016/j.ab.2012.12.008

Authors


Corinne Laclie

corinne.laclie@inrae.fr

Affiliation : CNRS, IFCE, INRAE, Université de Tours, UMR Physiologie de la Reproduction et des Comportements, Laboratoire Phénotypage Endocrinologie, F - 37380, Nouzilly, France

Country : France


Dominique Gennetay

Affiliation : CNRS, IFCE, INRAE, Université de Tours, UMR Physiologie de la Reproduction et des Comportements, Laboratoire Phénotypage Endocrinologie, F - 37380, Nouzilly, France

Country : France


Maïlys Gaudet

Affiliation : CNRS, IFCE, INRAE, Université de Tours, UMR Physiologie de la Reproduction et des Comportements, Laboratoire Phénotypage Endocrinologie, F - 37380, Nouzilly, France

Country : France


Anne-Lyse Lainé

Affiliation : CNRS, IFCE, INRAE, Université de Tours, UMR Physiologie de la Reproduction et des Comportements, Laboratoire Phénotypage Endocrinologie, F - 37380, Nouzilly, France

Country : France

Attachments

No supporting information for this article

Article statistics

Views: 17