Blood glucose curves in the management for diabetic patients have several limitations including intermittent assessment of blood glucose concentration, hospitalization, patient restraint, and repeated phlebotomy. The aim of this study was to apply and evaluate a wireless continuous glucose monitoring system (CGMS) in healthy dogs. Subcutaneous interstitial glucose concentrations in 7 dogs were continuously monitored and recorded by wireless CGMS. During induced hyperglycemia, the interstitial glucose concentrations were compared with whole blood glucose concentrations measured by glucometer and serum glucose concentrations measured by automated chemistry analyzer, respectively. There were no significant differences among interstitial, whole blood and serum glucose concentrations. The interstitial glucose concentrations had a good correlation to serum glucose concentrations. The real-time wireless CGMS is a valuable tool for monitoring system of glucose concentrations in dogs. Use of the CGMS for diabetic patients will provide accurate information over traditional blood glucose curves.