Mitral regurgitation (MR) resulting from myxomatous mitral valve disease (MMVD) is a prevalent condition in dogs, particularly smaller breeds like Cavalier King Charles Spaniels (CKCSs) and Chihuahuas (CHHs). An accurate assessment of MR severity is essential for effective treatment and disease monitoring, yet a standardized method has yet to be established. In this retrospective study, we evaluated 124 client-owned dogs diagnosed with MMVD, including 64 CKCSs and 60 CHHs. Dogs were categorized into three stages: asymptomatic (B1), remodeled (B2), and congestive heart failure (CHF, C). The MR severity was quantified using the proximal isovelocity surface area (PISA) method, specifically focusing on the PISA radius (PISA-r). The PISA-r measurements exhibited significant increases across disease stages and demonstrated strong correlations with echocardiographic parameters (ranging from 0.83 to 0.94), including the left atrial size and left ventricular internal diameter. The receiver operating characteristic (ROC) curve analysis revealed the high sensitivity and specificity of PISA-r in classifying disease stages, establishing optimal cut-off values. The method displayed excellent repeatability (interobserver variability: 0.95) and reproducibility (intraobserver variability: 0.97). In conclusion, the PISA method, specifically PISA-r, was reliable for assessing MR severity in dogs with MMVD. By simplifying the diagnosis and management of MR, this research can potentially improve the life and management of MMVD-affected dogs.