A time-resolved immunofluorometric assay for porcine C-reactive protein quantification in whole blood

摘要

A time-resolved immunofluorometric assay (TR-IFMA) for C-reactive protein (CRP) determination in whole blood of pigs was developed and validated. CRP was isolated from porcine acute-phase serum by affinity chromatography on agarose, coupled with phosphorylethanolamine and polyclonal antibodies to porcine CRP were purified from antiserum raised in sheep immunized with porcine CRP. Intra- and inter-assay coefficients of variation (CVs) were in the range 3.13-7.19% and 7.06-15.66%, respectively, showing good precision. The assay measured the CRP values in a proportional and linear manner (r = 0.99); additionally, CRP concentrations measured in whole blood by the present TR-IFMA and in serum by an established immunoturbidimetric assay were highly correlated (R2 = 0.97). The limit of detection of the method was 0.0028 mg/L. Significantly lower CRP concentrations were observed after 7 days of sample storage at 4℃. The injection of turpentine oil caused a significant increase in CRP concentrations and significantly higher CRP concentrations were observed in pigs with pathological processes compared to healthy animals.