Validation of commercial ELISAs for quantifying anabolic growth factors and cytokines in canine ACD-A anticoagulated plasma. (March 2017)

• Record Type: Journal Article
• Authors: Birdwhistell, Kate; Basinger, Robert; Hayes, Brian; Norton, Natalie; Hurley, David J.; Franklin, Samuel P.
• Is Part Of: Journal of veterinary diagnostic investigation. Volume 29:Number 2(2017:Mar.)
• Page Start: 143
• Page End: 147
• Publication Date: 2017-03
• Subjects: Anabolic growth factors -- dogs -- enzyme-linked immunosorbent assay -- inflammatory cytokines -- platelet-rich plasma; Veterinary medicine -- Diagnosis -- Periodicals; 636.0896075
• DOI: 10.1177/1040638717690186
• Languages: English
• ISSNs: 1040-6387

Abstract

Platelet-rich plasma has been studied extensively in dogs, but validation of enzyme-linked immunosorbent assays (ELISAs) for quantifying anabolic growth factors and inflammatory cytokines in canine plasma prepared with citrate-based anticoagulants is not available. We performed a validation of commercial ELISAs for transforming growth factor–beta 1 (TGF-β1), platelet-derived growth factor–BB (PDGF-BB), vascular endothelial growth factor (VEGF), tumor necrosis factor–alpha (TNF-α), and interleukin–1 beta (IL-1β) for use with canine plasma prepared with acid–citrate–dextrose, solution A (ACD-A). Platelet-poor plasma (PPP) anticoagulated with ACD-A as well as PPP anticoagulated with ACD-A and spiked with the relevant canine recombinant proteins were evaluated with each ELISA to calculate the efficiency of spike recovery. Replicates of the spiked PPP were also assessed in 2 additional assays to quantify intra-assay and interassay precision. The efficiency of spike recovery was within 75–125% of the expected concentration for the TGF-β1, PDGF-BB, and VEGF ELISAs. The intra- and interassay variability were <25% for the TGF-β1, PDGF-BB, VEGF, and TNF-α ELISAs. The TGF-β1, PDGF-BB, and VEGF ELISAs demonstrate acceptable efficiency of spike recovery and intra- and interassay variability, whereas the TNF-α and IL-1β ELISAs did not meet industry standards of performance with ACD-A anticoagulated canine plasma.