Development and validation of a novel competitive ELISA for the detection of serum amyloid A in pigs

• Published in: The veterinary journal (1997) 2011-11, Vol.190 (2), p.e7-e11
• Main Authors: Soler, L., Gutiérrez, A., Martínez-Subiela, S., Cerón, J.J.
• Format: Article
• Language: English
• Subjects: Acute phase protein; acute phase proteins; Acute phase response; Acute-Phase Reaction - blood; Acute-Phase Reaction - etiology; Acute-Phase Reaction - veterinary; amyloid; Animals; blood serum; Competitive ELISA; correlation; detection limit; enzyme-linked immunosorbent assay; Enzyme-Linked Immunosorbent Assay - methods; Enzyme-Linked Immunosorbent Assay - veterinary; Escherichia coli; Life Sciences; Limit of Detection; Linear Models; Male; Pig; polyclonal antibodies; Porcine Reproductive and Respiratory Syndrome - blood; Rabbits; Recombinant Proteins - analysis; Recombinant Proteins - metabolism; Serum amyloid A (SAA); Serum Amyloid A Protein - analysis; Serum Amyloid A Protein - metabolism; Swine
• ISSN: 1090-0233; 1532-2971
• DOI: 10.1016/j.tvjl.2011.02.016

The objective of this study was to develop a novel competitive ELISA to measure the acute phase protein serum amyloid A (SAA) in pigs using species-specific reagents. Polyclonal antibodies were produced in rabbits immunised with recombinant porcine SAA (rSAA) expressed in Escherichia coli. Both the rSAA and polyclonal antibodies were used to develop a novel competitive assay that was analytically and clinically validated. This assay had within- and between-run coefficients of variation of 8.6% and 25%, respectively, and demonstrated a high level of accuracy as determined by linearity-under-dilution (correlation coefficient, r=0.965). The analytical and functional limits of detection were 3.3 and 105.02mg/L, and the upper and lower quantification limits were 66.9 and 2.8mg/L, respectively. Statistically significant differences (P