The purpose of this study was to evaluate the diagnostic performance of a hand-held electronic glucometer (Precision Xtra; Abbott Diabetes Care Inc., Mississauga, ON, Canada) for cow-side use in dairy cattle. This device has been validated for measuring blood concentrations of β-hydroxybutyrate in dairy cows. This study was designed to assess the accuracy of whole-blood glucose measurements from the glucose meter relative to a reference chemical analyzer in a diagnostic laboratory. Duplicate samples were taken from the same cows at the same time, into blood tubes with either the glycolysis-inhibiting preservative sodium fluoride (NaF) or without preservative. Glucometer readings were taken on whole blood with no preservative, and laboratory measurements were conducted on serum preserved with NaF. Blood samples were collected from cows between 3 wk before and 5 wk after calving, including during a glucose tolerance test conducted 1 wk before expected calving. Passing-Bablok and Bland-Altman data analyses were used to evaluate the performance of the glucometer relative to the laboratory results. A strong correlation was observed in 709 samples from 81 cows between the hand-held meter and serum from samples preserved with NaF (R2=0.95). Overall, 96% of measurements with the glucometer fell within the 95% confidence limits of analysis in the laboratory, although at higher-than-physiologic glucose concentrations (>5.2mmol/L) the glucometer tended to overestimate. The hand-held glucometer appears suitable for rapid measurement of glucose under field conditions in dairy cattle.