Vet Med - Czech, 2007, 52(1):19-28 | DOI: 10.17221/2009-VETMED

Development of an ELISA-based kit for the on-farm determination of progesterone in milk

R. Simersky1,2, J. Swaczynova1,2, D.A. Morris1,2, M. Franek3, M. Strnad1,2
1 Laboratory of Growth Regulators, Palacky University, Olomouc, Czech Republic
2 Institute of Experimental Botany of the Academy of Sciences of the Czech Republic, Olomouc, Czech Republic
3 Veterinary Research Institute, Brno, Czech Republic.

Effective reproductive management is a crucial concern in the dairy industry and missed oestrus and late recognition of unsuccessful insemination can lead to substantial financial loss. Progesterone levels change in a predictable manner with progress of the oestrus cycle and with the onset of pregnancy. We report here the development of a simple ELISA test for the measurement of progesterone levels in bovine milk. The method is based on the use of polyclonal antibodies raised against 11α-hydroxyprogesterone-hemisuccinate and competition for antibody binding sites between free progesterone and horseradish peroxidase-labelled progesterone. Binding is quantified by means of the colour reaction between horseradish peroxidase and 3,3',5,5'-tetramethylbenzidine. The method has proved to be a cost effective, sensitive and easy to perform alternative to currently available methods of oestrus and pregnancy detection. The practical feasibility of the method has been proven under dairy farm conditions.

Keywords: bovine milk; ELISA; oestrus; progesterone; reproduction

Published: January 31, 2007  Show citation

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Simersky R, Swaczynova J, Morris DA, Franek M, Strnad M. Development of an ELISA-based kit for the on-farm determination of progesterone in milk. Vet Med - Czech. 2007;52(1):19-28. doi: 10.17221/2009-VETMED.
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    References

    1. Abraham G.E. (1969): Solid-phase radioimmunoassay of estradiol-17β. Journal of Clinical Endocrinology and Metabolism, 29, 866-870. Go to original source... Go to PubMed...
    2. Allen R.M., Redshaw M.R. (1978): The use of homologous and heterologous 125I-radioligands in the radioimmunoassay of progesterone. Steroids, 32, 467. Go to original source... Go to PubMed...
    3. Arnstadt K.I., Cleere W.F. (1981): Enzyme-immunoassay for determination of progesterone in milk from cows. Journal of Reproduction and Fertility, 62, 173-180. Go to original source... Go to PubMed...
    4. Beale M.H. (1999): Immunological methods in plant hormone research. In: Hooykaas P.J.J., Hall M.A., Libbenga K.R. (eds.): Biochemistry and Molecular Biology of Plant Hormones. Elsevier, Amsterodam. 61-84. Go to original source...
    5. Booth J.M., Davies J., Holdsworth R.J. (1979): Use of the milk progesterone for pregnancy determination. British Veterinary Journal, 135, 478-488. Go to original source... Go to PubMed...
    6. Claycomb R.W., Delwiche M.J., Munro C.J., BonDurant R.H. (1998): Rapid enzyme immunoassay for measurement of bovine progesterone. Biosensors & Bioelectronics, 13, 1165-1171. Go to original source... Go to PubMed...
    7. Cleere W.F., Gosling J.P., Morris M.C., Charleton M.F., Moloney B.T., Fottrell P.F., Sreenan J.M. (1984): A high performance, high throughput enzymeimmunoassay for the analysis of progesterone in plasma or milk. Irish Veterinary Journal, 39, 6-14.
    8. Dobson H., Midmer S.E., Fitzpatrick R.J. (1975): Relationship between progesterone concentrations in milk and plasma during the bovine oestrus cycle. Veterinary Record, 10, 222-223. Go to original source... Go to PubMed...
    9. Dray F., Andrieu J.M., Renaud F. (1975): Enzyme immunoassay of progesterone at the picogram level using β-galactosidase as label. Biochimica et Biophysica Acta, 403, 131-138. Go to original source... Go to PubMed...
    10. Elder P.A., Yeo K.H.J., Lewis J.G., Clifford J.K. (1987): An enzyme-linked immunosorbent assay (ELISA) for plasma progesterone: immobilised antigen approach. Clinica Chimica Acta, 162, 199-206. Go to original source... Go to PubMed...
    11. Erlanger B.F. (1967): The preparation of steroid-protein conjugates to elicit antihormonal antibodies. Methods in Immunology and Immunochemistry, 1, 144-150.
    12. Erlanger B.F., Borek F., Beiser S.M., Liebermann S. (1957): Steroid-protein conjugates. I. Preparation and characterization on conjugates of bovine serum albumin with testosterone and with cortisol. Journal of Biological Chemistry, 228, 713-727. Go to original source...
    13. Friggens N.C., Chagunda M.G.G. (2005): Prediction of the reproductive status of cattle on the basis of milk progesterone measures: model description. Theriogenology, 64, 155-190. Go to original source... Go to PubMed...
    14. Gowan E.W., Etches R.J. (1979): Solid-phase radioimmunoassay for progesterone and its application to pregnancy diagnosis in the cow. Theriogenology, 12, 327-343. Go to original source...
    15. Gross H., Bilk L. (1968): Zur reaktion von N-hydroxysuccinimide mit dicyklohexylcarbodiimid. Tetrahedron, 24, 6935-6939. Go to original source...
    16. Hansel W., Concannor P.W., Lukaszewska J.H. (1973): Corpora lutea of large domestic animals. Biology of Reproduction,8, 222-245. Go to original source...
    17. Harlow E., Lane D. (1988): Antibodies - A laboratory manual. Cold Spring Harbor Laboratory, USA.
    18. Heap R.B., Gwyn M., Laing J.A., Walters D.E. (1973): Pregnancy diagnosis in cows: changes in milk progesterone concentration during the oestrus cycle and pregnancy measured by a rapid radioimmunoassay. Journal of Agricultural Science, 81, 151-157. Go to original source...
    19. Holdsworth R.J., Chaplin V.M., Booth J.M. (1979): Radioimmunassay of progesterone in milk: development of techniques for large scale use as a test of pregnancy. British Veterinary Journal, 135, 470-477. Go to original source... Go to PubMed...
    20. Hruska K. (1996): Milk progesterone determination in dairy cows. Reproduction in Domestic Animals, 31, 483-485.
    21. Hruska K., Veznik Z. (1983): Significance of progesterone determination in milk. Endocrinologia Experimentalis, 17, 207-211. Go to PubMed...
    22. Hruska K., Franek M., Bursa J., Kalab P. (1983): Radioimmunoassay of progesterone in cow milk. Endocrinologia Experimentalis, 17, 213-218. Go to PubMed...
    23. Kyle B.L., Kennedy A.D., Small J.A. (1997): Measurement of vaginal temperature by radiotelemetry for the prediction of estrus in beef cows. Theriogenology, 49, 1437-1449. Go to original source... Go to PubMed...
    24. Laing J.A., Heap R.B. (1971): The concentration of progesterone in milk of cows during the reproductive cycle. British Veterinary Journal, 127, 19-22. Go to original source... Go to PubMed...
    25. Laing J.A., Eastman S.A.K., Bouflower J.C. (1979): The use of progesterone concentrations in milk and plasma for pregnancy diagnosis in cattle. British Veterinary Journal, 135, 204-209. Go to original source... Go to PubMed...
    26. Landon J., Crookall J., McGregor A. (1975): Enzyme-labelled immunoassays for steroids. In: Cameron E.H.D., Hillier S.G., Griffiths K. (eds.): Steroid Immunoassay. Alpha Omega, Cardiff. 183-188.
    27. Lucy M.C. (2001): Reproductive loss in high-producing dairy cattle: Where will it end? Journal of Dairy Science, 84, 1277-1293. Go to original source... Go to PubMed...
    28. Maatje K., Rossing W. (1976): Detecting oestrus by measuring milk temperatures of dairy cows during milking. Livestock Production Science, 3, 85-89. Go to original source...
    29. Midgley A.R. Jr., Niswender G.D. (1970): Radioimmunoassay of steroids. Acta Endocrinologica, 147, 320-328. Go to original source... Go to PubMed...
    30. Nakao T., Sugihashi A., Saga N., Tsunoda N., Kawata K. (1983): Use of milk progesteroneenzyme immunoassay for differential diagnosis of follicular cyst, luteal cyst and cystic corpus luteum in cows. American Journal of Veterinary Research, 44, 888-890.
    31. Nebel R.L. (1988): On-farm milk progesterone tests. Journal of Dairy Science, 71, 1682-1690. Go to original source... Go to PubMed...
    32. Oltenacu P.A., Ferguson J.D., Lednor A.J. (1990): Economic evaluation of pregnancy diagnosis in dairy cattle: A decision analysis approach. Journal of Dairy Science, 73, 2826-2831. Go to original source... Go to PubMed...
    33. O'Sullivan M.J., Bridges J.W., Marks V. (1979): Enzyme immunoassay: a review. Annals of Clinical Biochemistry, 16, 221. Go to original source... Go to PubMed...
    34. Prakash B.S., Meyer H.H.D., Van de Wiel D.F.M. (1988): Sensitive enzyme immunoassay of progesterone in skim milk using second-antibody technique. Animal Reproduction Science, 16, 225-235. Go to original source...
    35. Rorie R.W., Bilby T.R., Lester T.D. (2002): Application of electronic estrus detection technologies to reproductive management of cattle. Theriogenology, 57, 137-148. Go to original source... Go to PubMed...
    36. Royal M.D., Darwash A.O., Flint A.P.E., Webb R., Woolliams J.A., Lamming G.E. (2000): Declining fertility in dairy cattle: Changes in traditional and endocrine parameters of fertility. Animal Science, 70, 487-501. Go to original source...
    37. Ruiz F.J., Oltenacu P.A., Smith R.D. (1992): Cost-benefit evaluation of on-farm milk progesterone testing to monitor return to cyclicity and to classify ovarian cysts. Journal of Dairy Science, 75, 1036-1043. Go to original source... Go to PubMed...
    38. Satake K. (1960): The spectrophotometric determination of amine, amino acid and peptide with 2,4,6-trinintrobenzene 1-sulfonic acid. Journal of Biochemistry, 47, 654-660. Go to original source...
    39. Sauer M.J., Foulkes J.A., O'Neill P.M. (1981): Direct enzymeimmunoassay of progesterone in bovine milk. Steroids, 38, 45-53. Go to original source... Go to PubMed...
    40. Sprecher D.J., Nebel R.L., Whittier W.D (1988): Predictive value of palpation per rectum vs milk and serum progesterone levels for the diagnosis of bovine follicular and luteal cysts. Theriogenology, 30, 701-710. Go to original source... Go to PubMed...
    41. Stott A.W., Veerkamp R.F.,Wassel T.R. (1999): The economics of fertitlity in the dairy herd. Animal Science, 68, 49-57. Go to original source...
    42. Strnad M., Hanus J., Vanek T., Kaminek M., Ballantine J.A., Fussel B., Hanke D.E. (1997): Meta-topolin, a highly active aromatic cytokinin from poplar leaves (Populus x canadensis Moench, cv. Robusta). Phytochemistry, 45, 213-218. Go to original source...
    43. Van de Wiel D.F.M., Koops W. (1986): Development and validation of an enzyme immunoassay for progesterone in bovine milk or blood plasma. Animal Reproduction Science, 10, 201-213. Go to original source...

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