A time-resolved immunofluorometric assay for porcine C-reactive protein quantification in whole blood.
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 (R(2)=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 degrees C. 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.
Martinez-Subiela
S
S
Department of Animal Medicine and Surgery, University of Murcia, 30100 Espinardo, Murcia, Spain.
Eckersall
P D
PD
Campbell
F M
FM
Parra
M D
MD
Fuentes
P
P
Ceron
J J
JJ
D000818:Animals; D001769:Blood; D002097:C-Reactive Protein / Q000032:analysis* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013552:Swine
D002097:C-Reactive Protein