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

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.