Related Experiment Video
Updated: May 9, 2026

Multiplexed Fluorometric ImmunoAssay Testing Methodology and Troubleshooting
Published on: December 12, 2011
Analytical Validation of an Automated Point-of-Care Immunoassay for the Measurement of Feline Pancreatic Lipase
Lisa Wetzel1, Joerg M Steiner2, Harry Cridge1
1Department of Small Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, Michigan, USA.
Background:
Pancreatic lipase assays are commonly utilized in the diagnostic approach to suspected pancreatitis in cats. The Vcheck fPL 2.0 assay is commercially available; however, analytical validation has yet to be reported in the peer-reviewed literature.
Objective:
To evaluate the analytic validity of a fluorescent immunoassay for the point-of-care quantification of feline pancreatic lipase immunoreactivity (Vcheck fPL 2.0 assay).
Methods:
Linearity was assessed via dilutional parallelism. Intra- and interassay variability was assessed by calculating coefficients of variation (%CV) across replicates on the same day and across multiple days, respectively. Bilirubin, hemoglobin, and intralipid (intralipid 30% IV emulsion, VWR, Radnor, PA, USA) were utilized to evaluate the potential effects of interfering substances on the assay.
Results:
Linearity was sub-optimal, with 12/15 diluted runs having an O/E ratio outside of the 80-120% target range. The mean %CV was 10.2% for intra-assay variability and 16.3% for interassay variability. Interfering substances had no significant effect on the results of the Vcheck fPL 2.0 assay (P = 0.888).
Conclusions:
The Vcheck fPL 2.0 appears to have poor linearity and suboptimal precision, limiting the accurate interpretation of results. Additional assay optimization should be considered prior to routine clinical use.

