Analytical validation of an ELISA for measurement of canine pancreas-specific lipase

Abstract

Background: The diagnosis of canine pancreatitis is challenging. Clinical presentation often includes nonspecific clinical signs, such as vomiting, anorexia, and abdominal discomfort. Increased serum lipase activity can be indicative of pancreatitis; however, it can also be increased with other conditions. An immunoassay for measurement of canine pancreas-specific lipase in canine serum that would be suitable for commercial application and provide rapid results would be beneficial.

Objective: The goal of this study was to validate the Spec cPL assay, a commercially available ELISA for the quantitative measurement of canine pancreas-specific lipase.

Methods: Dynamic range, dilutional linearity, precision, interfering substances, assay stability, and reproducibility were investigated for analytical validation. The method was compared with the reference assay, canine pancreatic lipase immunoreactivity (cPLI), and included evaluation of a sample population of dogs and bias.

Results: Analytical validation showed a dynamic range of 36-954 μg/L; good precision (intra- and interassay coefficient of variation <12%); absence of interference from lipid, hemoglobin, or bilirubin; 12-month kit stability; and good reproducibility. Method comparison showed a positive bias relative to the cPLI reference method; however, the bias can be accommodated by adjustment of decision limits. The upper limit of the reference interval for Spec cPL was determined to be 216 μg/L based on the upper 97.5th percentile of results from 93 clinically healthy, kennel-housed dogs.

Conclusions: Validation data demonstrated that the Spec cPL assay provides reproducible results for canine pancreas-specific lipase. A readily available assay for measurement of this enzyme allows broader clinical utilization of this analytical tool, generating timely results to aid in the diagnosis of canine pancreatitis.