Comparison of 3 corneal cytology collection methods for evaluating equine ulcerative keratitis: Cytobrush, kimura platinum spatula, and handle edge of scalpel blade

PURPOSE To compare corneal cytology samples from three common sampling techniques: cytobrush (CB), Kimura platinum spatula (KS), and the handle edge of a scalpel blade (SB). METHODS Equine patients presenting to the University of Florida College of Veterinary Medicine with ulcerative keratitis were included. Following diagnosis of corneal ulcer and sampling for microbial culture, two cytology samples per technique were collected with sterile CB, KS, and SB in a randomized order. Cytologic evaluation was performed by two observers masked to collection method. Objective measures of sample cellularity, quality, distribution, and identification of infectious organisms were recorded per 10 monolayer cell populations using 50× magnification with oil immersion which were compared to culture results. Variables were compared using ANOVA with Student's t test when appropriate and Cohen's kappa (k) to evaluate inter- and intra-observer agreement (IOA) between observers and techniques. RESULTS Twenty equine patients (120 samples) were included. The IOA between observers was substantial (k = 0.75 ± 0.06) for cytological parameters. SB provided the most cellular samples (P < .01). There was a trend toward agreement (k = 0.12 ± 0.16) in technique for sample quality (P = .08). CB and SB had significantly poorer cell distribution than KS (P < .05). Infection was confirmed in 12 of 20 patients with SB and CB techniques having a significantly higher diagnostic yield than KS (P < .05) and was most consistent with infection confirmed on culture. CONCLUSIONS The SB provided the most diagnostic samples but all three techniques are clinically useful in evaluating equine ulcerative keratitis.