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

S Martinez-Subiela, P D Eckersall, Fiona Margaret Campbell, M D Parra, P Fuentes, J J Ceron

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    Abstract

    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.
    Original languageEnglish
    Pages (from-to)171-176
    Number of pages6
    JournalLuminescence
    Volume22
    Issue number3
    Early online date29 Jan 2007
    DOIs
    Publication statusPublished - May 2007

    Keywords

    • animals
    • blood
    • C-reactive protein
    • electrophoresis, polyacrylamide gel
    • fluorescent antibody technique
    • reproducibility of results
    • sensitivity and specificity
    • swine
    • TR-IFMA
    • whole blood
    • pigs

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  • Cite this

    Martinez-Subiela, S., Eckersall, P. D., Campbell, F. M., Parra, M. D., Fuentes, P., & Ceron, J. J. (2007). A time-resolved immunofluorometric assay for porcine C-reactive protein quantification in whole blood. Luminescence, 22(3), 171-176. https://doi.org/10.1002/bio.945