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Collection, Isolation, and Flow Cytometric Analysis of Human Endocervical Samples
Published on: July 6, 2014
A novel cytologic sampling technique to diagnose subclinical endometritis and comparison of staining methods for
O B Pascottini1, P Dini1, M Hostens1
1Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
Abstract:
The present article describes a study of the diagnosis of subclinical endometritis in dairy cows having two principal aims: first, to validate a novel technique for taking endometrial cytology samples to diagnose subclinical endometritis in dairy cows. Second, to compare the percentage of polymorphonuclear cells (PMNs) in cytology samples stained with Diff-Quik versus a specific staining method for PMNs, naphthol-AS-D-chloroacetate-esterase (CIAE). In the first experiment, Holstein-Friesian cows (n = 204) were used to take two cytology samples at the same time using the conventional cytobrush (CB) and the new cytotape (CT). Both devices were assembled within the same catheter allowing sampling at the same time, and approximately at the same location. Cytotape consisted of a 1.5-cm piece of paper tape rolled on the top of an insemination catheter covered with a double guard sheet. Parameters used to evaluate both methods were: PMNs percentage, total cellularity, quality of the smears, and red blood cell contamination. The concordance correlation coefficient analysis was used to assess agreement between continuous and Pearson chi-square tests for categorical variables. Agreement between the percentage of PMNs in both methods was good ρ = 0.84 (0.79, 0.87) with a minor standard error of 2%. Both methods yielded similar total cellularity (P = 0.62). Cytotape yielded better quality smears with more intact cells (P < 0.01) while samples that were taken by CB were more likely to be bloody (P < 0.01). Hence, CT and CB methods yielded smears with a similar PMNs percentage and a total number of cells, but CT provided smears with higher quality and significantly less blood contamination. For the second experiment, 114 duplicate cytology slides were stained using both Diff-Quik and CIAE. Agreement between PMNs percentage in both staining techniques was good ρc = 0.84 (0.78, 0.89) with a standard error of only 2%. Hence, Diff-Quik was confirmed as an easy, fast, and high-quality staining technique, which can be routinely used to stain endometrial cytology samples satisfactorily.
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