The aim of the current study was to determine the sensitivity and specificity of serum canine pancreatic elastase-1 (cPE-1) for the diagnosis of pancreatitis in dogs. The study was prospective, assessing dogs presenting with clinical signs similar to pancreatitis. Sixty-one dogs were recruited (49 with pancreatic disease and 12 with non-pancreatic disease). There was no significant difference in serum cPE-1 between dogs with pancreatic disease and non-pancreatic disease. However, there was a significant difference in serum cPE-1 between severe acute pancreatitis and non-pancreatic disease. A cut-off value for serum cPE-1 > 17.24 ng/ml resulted in sensitivity of 61.4% and specificity of 91.7% for diagnosis of all types of pancreatic disease. The sensitivity rose to 65.85% and 78.26% for the diagnosis of all types of acute pancreatitis and severe acute pancreatitis, respectively. Serum cPE-1 is more sensitive at diagnosing severe acute pancreatitis than chronic or mild acute pancreatitis, and has a high positive likelihood ratio. Dogs with chronic pancreatitis tended to have lower serum cPE-1 concentration, suggesting decreased exocrine function.