Development and validation of a novel competitive ELISA for the detection of serum amyloid A in pigs |
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| Authors: | Soler L Gutiérrez A Martínez-Subiela S Cerón J J |
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| Institution: | Department of Animal Medicine and Surgery, University of Murcia, Murcia, Spain. |
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| Abstract: | 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<0.0001) were found between the concentrations of SAA in healthy and diseased pigs. This novel assay precisely and sensitively measures SAA levels in pigs and will facilitate the more accurate assessment and study of the acute phase response in this species. |
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