Accuracy of a rapid enzyme-linked immunosorbent assay to measure progesterone in mares.
The aim of this study was to validate an enzyme-linked immunosorbent assay (ELISA) for the measurement of progesterone (P4) in mares. Specifically, the objectives were as follows: (1) to determine the specificity and sensitivity of the ELISA test for determination of P4, (2) to measure the potential agreement between the 2 people performing the test, and 3) to evaluate the effect of time on the outcome. Ten mares were sampled on the day before ovulation (D-1), and on days 1 (D1), 3 (D3), and 5 (D5) following ovulation, during the reproductive season. While mares were cycling regularly, estrus was induced by the injection of 5 mg of prostaglandin (PGF2) and monitored starting on the 4th day by daily transrectal palpation and ultrasonography to determine the time of ovulation. Blood was collected and all samples (n=96) were assayed for P4 by a semiquantitative ELISA, by chemiluminescent immunoassay, and by radioimmunoassay (RIA). Based on the RIA, values of P4 on D-1, D1, D3, and D5 were significantly different (P < 0.0001) with mean and standard deviation(s) of 0.004, s = 0.52; 2.05, s = 2.58; 8.37, s = 4.17; and 12.76, s = 4.00 ng/mL respectively. The sensitivity and specificity of the semiquantitative assay were 94% and 95%, respectively for the lowest values of P4 (< 1.0 ng/mL). The value of kappa was 0.90 between 2 individuals performing the test. In conclusion, these results suggest that the semiquantitative test may be used reliably and economically to evaluate P4 levels in equine plasma in the clinical
Relave
Fabien
F
Department of Clinical Sciences, University of Montreal, 3200 rue Sicotte, Saint-Hyacinthe.
Lefebvre
Réjean C
RC
Beaudoin
Sandra
S
Price
Christopher
C
D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards / Q000662:veterinary*; D004971:Estrus / Q000097:blood / Q000502:physiology; D005260:Female; D006736:Horses / Q000097:blood*; D007118:Immunoassay / Q000379:methods / Q000592:standards / Q000662:veterinary; D010060:Ovulation / Q000097:blood / Q000502:physiology; D011247:Pregnancy; D011270:Pregnancy, Animal / Q000097:blood / Q000502:physiology; D011374:Progesterone / Q000097:blood*; D011863:Radioimmunoassay / Q000379:methods / Q000592:standards / Q000662:veterinary; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors