Abstract
Simple SummaryAmong sepsis biomarkers, procalcitonin resulted to be a specific indicator of bacterial infection or severity of infection, and to be a good control of the success of a therapeutic procedure. The clinical studies on the relevance of procalcitonin as a sepsis predictor in veterinary patients are few, likely due to the total absence of validated assays. For this reason, this study aimed to investigate commercial ELISA kits for the detection of canine and equine procalcitonin. Validation was performed evaluating linearity, limits of detection (LOD), recovery, and intra-assay and inter-assay variability; furthermore, clinical samples were analyzed. The results of the present study demonstrate that the human PCT ELISA kit is suitable to detect equine procalcitonin with a LOD of 56 ng/mL, and the canine recombinant PCT ELISA kit can be used to measure canine procalcitonin in plasma samples, showing an intra-assay and inter-assay coefficient of variation less than 20% and a LOD of 11 pg/mL.In human medicine, procalcitonin (PCT), the precursor of calcitonin, is used for the rapid identification of the origin and severity of sepsis. In veterinary medicine, PCT has been studied in horses, cattle, and dogs, but the use of PCT in diagnostic and/or prognostic settings is not possible because of the lack of validated assays to obtain reference ranges. The aim of the present study was the investigation of commercially available ELISA kits for the detection of canine and equine PCT in plasma samples. Validation of the ELISA kits was performed by using species-specific recombinant proteins spiked both in plasma and buffer samples; linearity, limit of detection (LOD), recovery, and intra-assay and inter-assay variability were calculated. Moreover, clinical samples obtained from sick and healthy animals were also analyzed with the tested kits. Canine PCT was measured with a recombinant canine and a canine PCT ELISA kit. Equine PCT was measured with an equine and a human ELISA PCT kit. Our data demonstrate that the canine recombinant PCT ELISA kit can be used to measure canine PCT in plasma samples, showing an intra-assay and inter-assay coefficient of variation less than 20% and a LOD of 11 pg/mL, whereas the present results do not support the use of the canine PCT ELISA kit. The human PCT ELISA kit is suitable to detect equine PCT with a LOD of 56 ng/mL, whereas the equine PCT ELISA kit did not detect recombinant equine PCT.
Highlights
Procalcitonin (PCT) is a precursor of the hormone calcitonin and consists of 116-amino acids, which are normally processed by a specific protease to calcitonin, katacalcin, and an N-terminal residue [1]
The aim of this study was to evaluate commercially available enzyme-linked immunosorbent assays (ELISA) kits targeted for dog and horse for the detection of their respective plasma PCT levels
Our Systemic Inflammatory Response Syndrome (SIRS) group was composed of dogs (n = 5) and horses (n = 5) with a positive SIRS score (≥2 criteria) plus diagnosis of bacterial infection in order to only include animals truly affected by SIRS due to bacteria (Table 2)
Summary
Procalcitonin (PCT) is a precursor of the hormone calcitonin and consists of 116-amino acids, which are normally processed by a specific protease to calcitonin, katacalcin, and an N-terminal residue [1]. PCT in humans is used to distinguish patients with severe bacterial infections from those with severe non-septic conditions, to guide and reduce the antimicrobial treatment, and as a prognostic parameter [3]. For these reasons, PCT is very important to rapidly diagnose sepsis, minimize morbidity and mortality, and decrease unnecessary antibiotic use. Several methods are available to detect the PCT concentration in human blood, mostly using chemiluminescence [4,5], immunofluorescence [6], immunoluminometry [7,8], and surface plasmon resonance biosensors [9,10]. The use of PCT in diagnostic and/or prognostic settings in veterinary species is not possible because of the lack of appropriate and validated analytical methods to measure
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