View More View Less
  • 1 University of Veterinary Medicine, István u. 2, H-1078 Budapest, Hungary
  • 2 University of Veterinary Medicine, István u. 2, H-1078 Budapest, Hungary
  • 3 National Public Health Centre, Budapest, Hungary
Open access

Twenty-two serum samples of healthy bitches were tested with the frozen and lyophilised version of the same ELISA kit (Quanticheck, Faculty of Veterinary Science, Budapest, Hungary). Samples were chosen on the basis of their progesterone (P4) concentrations, which were between 1.00 and 20.00 ng/mL. As it is well known, this range has the highest clinical relevance in ovulation diagnosis. Both types of microplates were read at 15-min intervals from the 15th until the 90th minute (min) of incubation, and the results were compared with those of frozen plates at 60 min of incubation as 100 percent. Lyophilised microplates gave on average 18 percent higher results than the frozen version at equal incubation times. The highest difference between lyophilised and frozen samples was observed at 45 and 60 min of incubation. Ninety-four percent of the reaction in the frozen microplate occurred in the first 15 min, and during the subsequent 30 min the reaction seemingly stopped. After the 45th min of incubation, this 94 percent increased to 108 percent in the subsequent 30 min, which remained the final approximate result at the end of the 90 min of incubation. In contrast to the frozen microplate, the measured concentration increased continuously in the lyophilised version and reached the highest level at the 60th min. The results of the lyophilised microplate reached the same level at 30 min of incubation as those of the frozen version at 60 min. In conclusion, a mechanical increase or decrease of the incubation time does not generate a linear change in the test results. This study demonstrated that the results of a series of samples collected from the same bitch cannot be compared if they are measured with different laboratory methods or different ELISA kits.

If the inline PDF is not rendering correctly, you can download the PDF file here.

  • Chapwanya, A., Clegg, T., Stanley, P. and Vaughan, L. (2008): Comparison of the Immulite and RIA assay methods for measuring peripheral blood progesterone levels in Greyhound bitches.. Theriogenology 70, 795799.

    • Search Google Scholar
    • Export Citation
  • Concannon, P. W. (2011): Reproductive cycles of domestic bitch.. Anim. Reprod. Sci. 124, 200210.

  • Crowther, J. R. (Ed.) (1995): ELISA Theory and Practice. Humana Press Inc., Totowa, New Jersey, USA.

  • Henning, D. and Nielsen, K. (1987): Peroxidase-labelled monoclonal antibodies for use in enzyme immunoassay.. J. Immunoassay 8, 297307.

    • Search Google Scholar
    • Export Citation
  • Hock, B., Rahman, M., Rauchalles, S., Dankwardt, A., Seifert, M., Haindl, S. and Kramer, K. (1999): Stabilisation of immunoassays and receptor assays.. J. Mol. Catal. B: Enzym. 7, 115124.

    • Search Google Scholar
    • Export Citation
  • Molnár, L., Ács, J., Csernus, V., Solti, L., Varró, R. and Huszenicza, Gy. (1984): Enzyme immunoassay (EIA) of progesterone. Proc. of the Symp. on the Analysis of Steroids, Szeged, Hungary.

    • Search Google Scholar
    • Export Citation
  • Munro, C. and Stabenfeldt, G. (1984): Development of a microtitre microplate enzyme immunoassay for the determination of progesterone.. J. Endocrinol. 10, 4149.

    • Search Google Scholar
    • Export Citation
  • Navarro, H., Zarco, L., Ducoing, A., Flores, G. and Valencia, J. (1990): Effect of time and temperature of incubation of heparinized caprine blood on concentrations of progesterone detected in plasma.. Theriogenology 33, 749755.

    • Search Google Scholar
    • Export Citation
  • Schumacher, M., Mattern, C., Ghoumari, A., Oudinet, J. P., Liere, P., Labombarda, F., Sitruk-Ware, R., De Nicola, A. F. and Guennoun, R. (2014): Revisiting the roles of progesterone and allopregnanolone in the nervous system: Resurgence of the progesterone receptors.. Prog. Neurobiol. 113, 639.

    • Search Google Scholar
    • Export Citation
  • Sibley, D. E. T., Ramsay, G., Lubrano, G. J. and Guilbault, G. G. (1993): Stability and reusability of enzyme-linked immunosorbent assay (ELISA) microplates.. Anal. Lett. 26, 16231634.

    • Search Google Scholar
    • Export Citation
  • Tahir, M. Z., Thoumire, S., Raffaelli, M., Grimard, B., Reynaud, K. and Chastant-Maillard, S. (2013): Effect of blood handling conditions on progesterone assay results obtained by chemiluminescence in the bitch.. Domest. Anim. Endocrin. 45, 141144.

    • Search Google Scholar
    • Export Citation
  • Thuróczy, J., Wölfling, A., Tibold, A., Balogh, L., Jánoki, Gy. A. and Solti, L. (2003): Effect of anticoagulants and sampling time on results of progesterone determination in canine blood samples.. Reprod. Domest. Anim. 38, 386389.

    • Search Google Scholar
    • Export Citation
  • Volkmann, D. H. (2006): The effects of storage time and temperature and anticoagulant on laboratory measurements of canine blood progesterone concentrations.. Theriogenology 66, 15831586.

    • Search Google Scholar
    • Export Citation

Monthly Content Usage

Abstract Views Full Text Views PDF Downloads
Aug 2020 0 6 0
Sep 2020 0 3 0
Oct 2020 0 4 7
Nov 2020 0 4 4
Dec 2020 0 2 3
Jan 2021 0 3 5
Feb 2021 0 0 0