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Analytical and bioanalytical chemistry2014; 406(22); 5507-5512; doi: 10.1007/s00216-014-7944-z

A new ELISA for the quantification of equine procalcitonin in plasma as potential inflammation biomarker in horses.

Abstract: In human medicine, procalcitonin (PCT) is a very common and well-established biomarker for sepsis. Even though sepsis is also a leading cause of death in foals and adult horses, up to now, no data about the role of equine PCT in septic horses has been available. Based on monoclonal antibodies targeted against human PCT, we report here the development of a sandwich ELISA for the quantification of equine PCT in equine plasma samples. The ELISA was characterized for intra- and interassay variance and a working range from 25 to 1,000 ng mL(-1) was defined as within this range; both intra- and interassay variances were below 15 %. The target recovery ranged between 73 and 106 %. The ELISA was used to determine the equine PCT concentration in 24 healthy and 5 septic horses to show the potential for clinical evaluation of equine PCT. Significantly different (P = 0.0006) mean equine PCT concentrations were found for the healthy control group and the sepsis group (47 and 8,450 ng mL(-1)).
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Publication Date: 2014-06-14 PubMed ID: 24928115DOI: 10.1007/s00216-014-7944-zGoogle Scholar: Lookup
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Summary

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The research article presents the development of a new Enzyme-linked immunosorbent assay (ELISA) aimed at measuring the levels of procalcitonin, a biomarker for inflammation, in horses. The study has demonstrated a clear difference in procalcitonin concentrations between healthy horses and those with sepsis.

Development of Sandwich ELISA for Equine PCT

  • The researchers developed a new sandwich ELISA using monoclonal antibodies targeted against human PCT, but designed for the measurement of equine PCT in horse plasma samples.
  • This method was chosen due to its high specificity and sensitivity, which are important characteristics for a reliable diagnostic tool.
  • The ELISA devised for this research has a working range from 25 to 1,000 ng mL(-1). Any readings within this range are considered reliable, and this helps in reducing errors in the test results.

Characterization and Variance

  • The researchers evaluated the new ELISA based on two main criteria: intra-assay variance (the variability of results within a single test run) and interassay variance (the variability of results between separate test runs).
  • Both intra- and interassay variances for this ELISA fell below 15%, indicating a high level of precision and consistency in the assay’s measurements.
  • Additionally, the target recovery, or the amount of the initial analyte recovered after the testing, ranged between 73 and 106%, showcasing the test’s accuracy.

Evaluation of Equine PCT in Healthy and Septic Horses

  • The researchers used the newly developed ELISA to establish the PCT concentration in a total of 29 horses: 24 who were healthy, and 5 who had a confirmed case of sepsis.
  • Significant differences in PCT concentrations were found between the healthy control group and the sepsis group. The mean PCT concentrations for the healthy group was notably lower (47 ng mL(-1)) compared to the sepsis group (8,450 ng mL(-1)). This result strongly suggests the utility of the developed ELISA for identifying sepsis in horses.

Conclusion

This research shows promise in the application of the new ELISA for the diagnosis of sepsis and potentially other inflammatory conditions in horses. The developed test exhibits a high level of precision, accuracy, and consistency, which are essential properties for any diagnostic tool. The identification of significantly high PCT concentrations in septic horses compared to healthy ones further reinforces the potential of this tool. Further research and clinical testing are needed to completely validate its utility.

Cite This Article

APA
Rieger M, Kochleus C, Teschner D, Rascher D, Barton AK, Geerlof A, Kremmer E, Schmid M, Hartmann A, Gehlen H. (2014). A new ELISA for the quantification of equine procalcitonin in plasma as potential inflammation biomarker in horses. Anal Bioanal Chem, 406(22), 5507-5512. https://doi.org/10.1007/s00216-014-7944-z

Publication

Analytical and bioanalytical chemistry
ISSN: 1618-2650
NlmUniqueID: 101134327
Country: Germany
Language: English
Volume: 406
Issue: 22
Pages: 5507-5512

Researcher Affiliations

Rieger, Martin
  • Research Unit Microbe-Plant Interactions, Department of Environmental Sciences, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), Ingolstädter Landstraße 1, 85764, Neuherberg, Germany, martin.rieger@helmholtz-muenchen.de.
Kochleus, Christian
    Teschner, Dana
      Rascher, Daniela
        Barton, Ann Kristin
          Geerlof, Arie
            Kremmer, Elisabeth
              Schmid, Michael
                Hartmann, Anton
                  Gehlen, Heidrun

                    MeSH Terms

                    • Animals
                    • Antibodies, Monoclonal / chemistry
                    • Biomarkers / blood
                    • Calcitonin / blood
                    • Calcitonin Gene-Related Peptide
                    • Enzyme-Linked Immunosorbent Assay / methods
                    • Female
                    • Horses
                    • Humans
                    • Inflammation
                    • Male
                    • Protein Precursors / blood
                    • ROC Curve
                    • Recombinant Proteins / chemistry
                    • Reproducibility of Results
                    • Sepsis / blood
                    • Sepsis / diagnosis

                    Citations

                    This article has been cited 17 times.
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