Development and validation of an enzyme-linked immunosorbent assay for feline trypsin-like immunoreactivity

Jörg M. Steiner, David A. Williams, Erik M. Moeller, Tonatiuh Melgarejo

Research output: Contribution to journalArticlepeer-review

Abstract

Objective - To develop and validate an ELISA for quantitative analysis of feline trypsin-like immunoreactivity (fTLI). Sample Population - Purified feline cationic trypsin (fCT) and rabbit anti-fCT antiserum; blood samples from 63 healthy cats. Procedures - A sandwich capture ELISA was developed, using anti-fCT antiserum purified by affinity chromatography that underwent biotinylation. Purified fCT was used for standards. The assay was validated by determination of sensitivity, working range, linearity, accuracy, precision, and reproducibility. A reference range was established by assaying serum samples from the 63 healthy cats. Results - Sensitivity was 1.23 μg/L; working range was 2 to 567 μg/L. Ratios of observed versus expected results for 4 samples tested at various dilutions ranged from 90.0 to 120.7%. Ratios of observed versus expected results for 5 samples spiked with various concentrations of fCT ranged from 82.0 to 101.8%. Intra-and inter-assay coefficients of variability ranged from 9.9 to 11.1% and from 10.2 to 21.7%, respectively. The reference range for serum fTLI measured with this ELISA was 12 to 82 μg/L. Conclusions and Clinical Relevance - Results suggest that an ELISA can be used to measure serum fTLI in cats. The ELISA was sufficiently sensitive, linear, accurate, precise, and reproducible for clinical use.

Original languageEnglish (US)
Pages (from-to)620-623
Number of pages4
JournalAmerican journal of veterinary research
Volume61
Issue number6
DOIs
StatePublished - Jun 2000
Externally publishedYes

ASJC Scopus subject areas

  • veterinary(all)

Fingerprint Dive into the research topics of 'Development and validation of an enzyme-linked immunosorbent assay for feline trypsin-like immunoreactivity'. Together they form a unique fingerprint.

Cite this